Phospholipases, nucleic acids encoding them and methods for preparing and using them

Abstract

PHOSPHOLIPASES, NUCLEIC ACIDS ENCODING THEM AND METHODS TO PREPARE AND USE THEM. The invention provides new polypeptides that have phospholipase activity, including, for example, phospholipase A, B, CED activity, patatin activity, phosphatidic acid phosphates ( PAP) and / or lipid acyl hydrolase (LAH) activity, nucleic acids encoding them and antibodies that bind to them. Industrial methods, for example, degumming of oil and products comprising use of these phospholipases are likewise provided.

Description

Descriptive Report of the Patent of Invention for "PHOSPHOLIPASES, NUCLEIC ACIDS ENCODING THEM AND METHODS FOR PREPARING AND USING THEM". Field of Invention The present invention generally relates to phospholipase enzymes, polynucleotides encoding the enzymes, methods of preparing and employing these polypeptide polynucleotides. In particular, the invention provides novel polypeptides having phospholipase activity, nucleic acids encoding them, and antibodies that bind to them. Industrial methods and products comprising the use of these phospholipases are also provided. background Phospholipases are enzymes that hydrolyze the ester bonds of phospholipids. Corresponding to their importance in the metabolism of phospholipids, these enzymes are widespread among prokaryotes and eukaryotes. Phospholipases affect the metabolism, construction and reorganization of biological membranes and are involved in signal cascades. Several types of phospholipases are known that differ in their specificity according to the position of the bond attached to the phospholipid molecule. Phospholipase A1 (PLA1) removes the 1-position fatty acid to produce free fatty acid and 1-lyso2-acylphospholipid. Phospholipase A2 (PLA2) removes the 2-position fatty acid to produce the free fatty acid and 1-acyl-2-lysophospholipid. PLA1 and PLA2 enzymes can be intra- or extra-cellular, membrane-bound or soluble. Intracellular PLA2 is found in almost every mammalian cell. Phospholipase C (PLC) removes the phosphate moiety to produce phosphate ester and 1,2-diacylglycerol. Phospholipase D (PLD) produces 1,2-diacylglycerophosphate and base group. PLC and PLD are important in cell fusion and signaling. PLD has been the dominant phospholipase in biocatalysis (see, for example, Godfrey, T. and West S. (1996) Industrial enzymology, 299-300, Stockton Press, New York). Patatins are another type of phospholipase, thought to work like a PLA (see, for example, Hirschberg HJ, et al., (2001), Eur J Biochem 268(19):5037-44). Common oil seeds such as soybeans, rapeseed, sunflower, rice bran oil, sesame and peanut are used as sources of oil and feedstock. In the oil extraction process, the seeds are mechanically and thermally treated. The oil is separated and divided from the flour by a solvent. Using distillation, the solvent is then separated from the oil and recovered. The oil is "degummed" and refined. The solvent content in the flour can be evaporated by heat treatment in a "desolventizing toaster," followed by drying and cooling the flour. After a solvent has been distilled off, the crude oil produced is processed into edible oil, employing special degumming and physical refinement procedures. It can be used as a feedstock for the production of fatty acids and methyl ester. The flour can be used for animal feed. Degumming is the first step in vegetable oil refining and is designed to remove contaminating phosphatides that are extracted with the oil but interfere with subsequent oil processing. These phosphatides are soluble in vegetable oil only in an anhydrous form and can be precipitated and removed if they were simply hydrated. Hydration is usually carried out by mixing a small proportion of water with substantially dry oil. Typically, the amount of water is 75% of the phosphatide content, which is typically 1 to 1.5%. Temperature is not highly critical, although separation of the hydrated gums is better when the oil viscosity is reduced to 50°C to 80°C. Many methods for degumming oil are currently employed. The oil degumming process can be enzymatically aided by employing phospholipase enzymes. Phospholipases A1 and A2 have been employed for oil degumming in various commercial processes, for example "ENZYMAX® degumming" (Lurgi Life Science Technologies GmbH, Germany). Phospholipase C (PLC) has also been considered for oil degumming because the phosphate moiety generated by its action on phospholipids is very water soluble and easy to remove and the diglyceride would remain with the oil and reduce the losses; see, for example, Godfrey, T. and West S. (1996) Industrial Enzymology, pp. 299-300, Stockton Press, New York; Dahlke (1998) "An enzymatic process for the physical refining of seed oils," Chem. Eng. Technol. 21:278-281; Clausen (2001) "Enzymatic oil degumming by a novel microbial phospholipase," Eur. J. Lipid Sci. Technol. 103:333-340. High phosphatide oils such as soybean, canola and sunflower are processed differently from other oils such as palm. Unlike the steam process or "physical refinement" for low-phosphatide oils, these high-phosphorus oils require special chemical and mechanical treatments to remove the phosphorus-containing phospholipids. These oils are typically chemically refined in a process that links the neutralization of free fatty acids to form soap and an insoluble gum fraction. The neutralization process is highly effective in removing free fatty acids and phospholipids, but this process also results in significant yield losses and quality sacrifices. In some cases, the high phosphatide crude oil is deslimed in a step that precedes caustic neutralization. This is the case for soybean oil used for lecithin, where the oil is first degummed by water or acid. Phytosterols (plant sterols) are members of the "tri-terpene" family of natural products, which include more than 100 different phytosterols and more than 4000' other types of triterpenes. In general, phytosterols are thought to stabilize plant membranes, with an increase in the sterol / phospholipid ratio leading to membrane rigidity. Chemically, phytosterols closely resemble cholesterol in structure and are thought to regulate membrane fluidity in plant membranes, like cholesterol in animal membranes. The main phytosterols are p-sitosterol, campesterol and stigmasterol. Others include stigmastanol (p-sitostanol), sitostanol, desmosterol, dihydrobrassicasterol, calinasterol, poriferasterol, clionasterol and brassicasterol. Plant sterols are important agricultural products for the nutritional and health industries. They are useful emulsifiers for manufacturing cosmetic ingredients and provide most steroid intermediates and precursors for pharmaceutical hormone production. Saturated phytosterol analogues and their esters have been suggested as effective cholesterol-lowering agents with cardiac health benefits. Plant sterols reduce serum cholesterol levels by inhibiting the absorption of cholesterol in the intestinal lumen and have immunomodulatory properties at extremely low concentrations, including enhanced cellular response of T lymphocytes and the cytotoxic capacity of natural killer cells against a cellular strain of cancer. Furthermore, its therapeutic effect has been demonstrated in clinical studies for the treatment of pulmonary tuberculosis, rheumatoid arthritis, control of HIV-infected patients and inhibition of immune tension in marathon runners. Plant sterol esters, also referred to as phytosterol esters, were approved as GRAS (Generally Recognized As Safe) by the US Food and Drug Administration (FDA) for use in margarine and spreads in 1999. In September 2000, the FDA similarly issued an interim rule allowing labeling of health claims of foods containing phytosterol esters. Consequently, fortification of foods with phytosterol esters is highly desired for consumer acceptance. Soybean oil is widely employed and is an important foodstuff, accounting for ~30% of oil production from seeds and fruits. Soybeans contain only 20% oil and extraction is normally completed using a solvent such as hexane on a commercial scale. The recognized quality of its oil and the nutritional value of the flour protein makes soy a primary oilseed. Prior to extraction, soybeans must be cleaned, cracked and chipped as efficient solvent extraction of oil requires that every oil cell be broken down to improve mass transfer. Cell walls mainly composed of cellulose, associated with hemicelluloses, pectic substances and lignin), can likewise be broken down by means of enzymes, to obtain a significant improvement in rates and yields of extraction. Diacylglycerol oil (DAG) is an edible oil containing 80% or more of DAG than natural fatty acids. Postprandial elevation of triglyceride in chylomicrons has been shown in humans to be markedly lower after ingestion of a DAG oil emulsion compared to a TAG oil with a similar fatty acid composition. In studies employing American women and men and Japanese men, long-term consumption of DAG oil promoted weight loss and reduced body fat. One study showed that replacing DAG oil with ordinary cooking oil reduces the incidence of obesity and similar risk factors. Summary of the Invention The invention provides recombinant or isolated nucleic acids comprising a nucleic acid sequence having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60 %, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for an exemplary nucleic acid of the invention, e.g., SEQ ID NO: 1, SEQ ID NO: 3, SEO ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, S EQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO : 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145 , SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 199, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, over a region of at least about 10, 15, 20, 25 , 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000 , 1050,1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, 2000, 2050, 2100, 2200, 2250, 2300 , 2350, 2400, 2450, 2500, or more residues, and in one aspect the nucleic acid encodes a polypeptide having at least one phospholipase (PL) activity, for example, a phospholipase (PL) activity. and phospholipase A, C or D, or any combination of phospholipase activity, for example a PL A, PL C and / or PL D activity as a multifunctional activity. In one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The invention provides recombinant or isolated nucleic acids comprising a nucleic acid sequence having at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or more, or full sequence identity (100%) for SEQ ID NO: 1 over a region of at least approx. of 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800 , 850 more consecutive residues, and in one aspect the nucleic acid encodes at least one polypeptide having a phospholipase (PL) activity, for example, a phospholipase A, C or D activity, or any combination of phospholipase activity, for example , a PL A, PL C and / or PL D activity as a cross-functional activity. In one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The invention provides recombinant or isolated nucleic acids comprising a nucleic acid sequence having at least 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for SEQ ID NO: 3 over a region of at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600 , 650, 700, 750, 800, 850 or more residues, and in one aspect the nucleic acid encodes at least one polypeptide having a phospholipase (PL) activity, for example, a phospholipase A, C or D activity, or any combination of phospholipase activity, for example a PL A, PL C and / or PL D activity as a multifunctional activity. In one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The invention provides recombinant or isolated nucleic acids comprising a nucleic acid sequence having at least 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for SEQ ID NO: 5 over a region of at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600 , 650, 700, 750, 800, 850 or more residues, and in one aspect the nucleic acid encodes at least one polypeptide having a phospholipase (PL) activity, for example, a phospholipase A, C or D activity, or any combination of phospholipase activity, for example a PL A, PL C and / or PL D activity as a multifunctional activity. In one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The invention provides recombinant or isolated nucleic acids comprising a nucleic acid sequence having at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99%, or more, or complete sequence identity (100%) for SEQ ID NO: 7 over a region of at least about 10, 15, 20 , 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850 or more residues, and in one aspect the nucleic acid encodes at least one polypeptide having a phospholipase (PL) activity, for example, a phospholipase A, C or D activity, or any combination of phospholipase activity, for example, a phospholipase activity. PL A, PL C and / or PL D as a cross-functional activity. In one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. In alternative aspects, the recombinant or isolated nucleic acid encodes a polypeptide comprising a sequence as mentioned in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO : 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132 , SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEO ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO: 174. aspect, these polypeptides have a phospholipase, e.g., a phospholipase A, B, C, or D activity, or any combination of phospholipase activity, e.g., a PL A, PL C, and / or PL D activity, as an activity. multifunctional. In one aspect, the sequence comparison algorithm is a BLAST algorithm, such as a version 2.2.2 BLAST algorithm. In one respect, the filtration setting is set to blastall -p blastp -d "nr pataa" - F F and all other options are set to default. In one aspect, the phospholipase activity comprises catalyzing hydrolysis of a glycerolphosphate ester bond (i.e., cleavage of glycerolphosphate ester bonds). Phospholipase activity may comprise catalyzing hydrolysis of an ester bond on a phospholipid in a vegetable oil. The vegetable oil phospholipid may comprise an oil seed phospholipid. The phospholipase activity may comprise a phospholipase C (PLC) activity; a phospholipase A (PLA) activity, such as a phospholipase A1 or phospholipase A2 activity; a phospholipase D (PLD) activity, such as a phospholipase D1 activity or a phospholipase D2 activity; a phospholipase B (PLB) activity, for example, a phospholipase and a lysophospholipase (LPL) activity or a phospholipase and a lysophospholipase-transacylase (LPTA) activity or a phospholipase and a lysophospholipase (LPL) activity and lysophospholipase-transacylase (LPTA) activity; or patatin activity, or a combination thereof. Phospholipase activity may comprise hydrolysis of a glycoprotein, for example as a glycoprotein found in a potato tuber. The phospholipase activity may comprise a patatin enzymatic activity. Phospholipase activity may comprise a lipid acyl hydrolase (LAH) activity. In one aspect, a phospholipase of the invention may have multifunctional activity, for example, a combination of one or more of the enzyme activities described herein, for example, a phospholipase of the invention may have PLC and PLA activity; PLB and PLA activity; PLC and PLD activity; PLC and PLB activity; PLB and patatin activity; PLC and patatin activity; PLD and PLA; PLD, PLA, PLB and PLC activity; or PLD, PLA, PLB, PLC and patatin activity; or, a phospholipase and a lysophospholipase (LPL) activity or a phospholipase and a lysophospholipase-transacylase (LPTA) activity or a phospholipase and a lysophospholipase (LPL) activity and a lysophospholipase-transacylase (LPTA) activity, or any combination of these. For example, in one aspect, a polypeptide of the invention is enzymatically active, but needs a lipase activity, e.g., needs any enzymatic activity that affects a neutral oil (triglyceride) fraction. It may be desirable to use such a polypeptide in a particular process, for example in a desizing process where it is important that the neutral oil fraction is not harmed (decreased, for example hydrolyzed). Thus, in one aspect, the invention provides a desizing process comprising using a polypeptide of the invention having a phospholipase activity, but not a lipase activity. In one aspect, the isolated or recombinant nucleic acid encodes a polypeptide having a phospholipase activity that is thermostable. The polypeptide can maintain phospholipase activity under conditions comprising a temperature range of from about 20°C to about 30°C, from about 25°C to about 40°C, from about 37°C to about 40°C. 95°C; from about 55°C to about 85°C, from about 70°C to about 95°C, or from about 90°C to about 95°C. In another aspect, the isolated or recombinant nucleic acid encodes a polypeptide having a phospholipase activity that is thermotolerant. The polypeptide can maintain phospholipase activity after exposure at a temperature in the range greater than 37 °C to about 95 °C or anywhere in the range greater than 55 °C to about 85 °C. In one aspect, the polypeptide maintains a phospholipase activity after exposure to a temperature in the range greater than 90°C to about 95°C at pH 4.5. The polypeptide can maintain phospholipase activity under conditions comprising about pH 8, pH 7.5, pH 7, pH 6.5, pH 6.0, pH 5.5, pH 5, or pH 4.5. The polypeptide can maintain phospholipase activity under conditions comprising a temperature range of from about 40°C to about 70°C. In one aspect, the recombinant or isolated nucleic acid comprises a sequence that hybridizes under stringent conditions to a sequence as mentioned in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO : 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145 , SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, wherein the nucleic acid encodes a polypeptide having a phospholipase activity. The nucleic acid can be at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650 , 700, 750, 800, 850 or residues in length or the full length of the gene or copy, with or without a signal sequence, as described herein. Stringent conditions can be highly severe, moderately severe or low-severity, as described here. Severe conditions may include a washing step, for example a washing step comprising a wash in 0.2X SSC at a temperature of about 65°C for about 15 minutes. The invention provides a nucleic acid probe for identifying a nucleic acid encoding a polypeptide with a phospholipase, for example, a phospholipase activity, wherein the probe comprises at least 10, 20, 30, 40, 50, 60, 70 , 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, or more, consecutive bases of a sequence of the invention, for example , a sequence as mentioned in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7, and the probe identifies the nucleic acid by ligation or hybridization. The probe may comprise an oligonucleotide comprising at least about 10 to 50, about 20 to 60, about 30 to 70, about 40 to 80, or about 60 to 100 consecutive bases of a sequence as mentioned in SEQ ID NO. : 1, SEQ ID NO: 3, SEQ ID NO: 5 and / or SEQ ID NO: 7. The invention provides a nucleic acid probe for identifying a nucleic acid encoding a polypeptide with a phospholipase, for example, a phospholipase activity, wherein the probe comprises a nucleic acid of the invention, for example, a nucleic acid having 450, 500, 550, 600, 650, 700, 750, 800, 850 or more consecutive residues; and, in one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The invention provides an amplification primer sequence pair for amplifying a nucleic acid encoding a polypeptide having a phospholipase activity, wherein the primer pair is capable of amplifying a nucleic acid comprising a sequence of the invention, or fragments or subsequences thereof. One or each member of the amplification primer sequence pair may comprise an oligonucleotide comprising at least about 10 to 50 consecutive bases of sequence, or about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more consecutive bases of sequence. The invention provides amplification primer pairs, wherein the primer pair comprises a first member having a sequence mentioned by about the first (5') 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more residues of a nucleic acid of the invention, and a second member having a sequence mentioned around the first (5') 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23, 24, or 25 or more residues from the complementary strand of the first member. The invention provides phospholipases generated by amplification, for example, polymerase chain reaction (PCR), employing an amplification primer pair of the invention. The invention provides methods for preparing a phospholipase by amplification, for example, polymerase chain reaction (PCR), employing an amplification primer pair of the invention. In one aspect, the amplification primer pair amplifies a nucleic acid from a library, for example, a gene library, such as an environmental library. The invention provides methods of amplifying a nucleic acid encoding a polypeptide having a phospholipase activity comprising amplifying a standard nucleic acid with an amplification primer sequence pair capable of amplifying a nucleic acid sequence of the invention, or fragments or subsequences thereof. The amplification primer pair may be an amplification primer pair of the invention. The invention provides expression cassettes comprising a nucleic acid of the invention or a subsequence thereof. In one aspect, the expression cassette may comprise nucleic acid that is operably linked to a promoter. The promoter may be a viral, bacterial, mammalian or plant promoter. In one aspect, the plant promoter may be a potato, rice, corn, wheat, tobacco or barley promoter. The promoter may be a constitutive promoter. The constitutive promoter may comprise CaMV35S. In another aspect, the promoter may be an inducible promoter. In one aspect, the promoter may be a tissue-specific promoter or a developmentally regulated promoter or an environmentally regulated. Thus, the promoter may be, for example, a seed-specific, a leaf-specific, a root-specific, a stem-specific or an abscission-induced promoter. In one aspect, the expression cassette may also comprise a plant or plant virus expression vector. The invention provides cloning vehicles comprising an expression cassette (e.g. a vector) of the invention or a nucleic acid of the invention. The cloning vehicle can be a viral vector, a plasmid, a phage, a phagemid, a cosmid, a phosmid, a bacteriophage or an artificial chromosome. The viral vector may comprise an adenovirus vector, a retroviral vector or an adeno-associated viral vector. The cloning vehicle may comprise a bacterial artificial chromosome (BAC), a plasmid, a bacteriophage P1-derived vector (PAC), a yeast artificial chromosome (YAC), or a mammalian artificial chromosome (MAC). The invention provides transformed cell comprising a nucleic acid of the invention or an expression cassette (e.g. a vector) of the invention, or a cloning vehicle of the invention. In one aspect, the transformed cell can be a bacterial cell, a mammalian cell, a fungal cell, a yeast cell, an insect cell, or a plant cell. In one aspect, the plant cell may be a cell of potato, wheat, rice, corn, tobacco or barley. The invention provides non-human transgenic animals comprising a nucleic acid of the invention or an expression cassette (e.g. a vector) of the invention. In one aspect, the animal is a mouse, rat, cow, sheep, or other mammal. The invention provides transgenic plants comprising a nucleic acid of the invention or an expression cassette (e.g. a vector) of the invention. The transgenic plant can be a corn plant, a potato plant, a tomato plant, a wheat plant, an oilseed plant, a rapeseed plant, a soybean plant, a rice plant, a plant of barley or a tobacco plant. The in- The invention provides transgenic seeds comprising a nucleic acid of the invention or an expression cassette (e.g. a vector) of the invention. The transgenic seed can be a corn seed, a wheat seed, an oilseed, a rapeseed (a canola plant), a soybean seed, a palm seed, a sunflower seed, a sesame seed, a seed of a peanut, rice or tobacco plant. The invention provides an antisense oligonucleotide comprising a nucleic acid sequence complementary to or capable of hybridizing under stringent conditions to a nucleic acid of the invention. The invention provides methods of inhibiting the translation of a phospholipase message in a cell comprising administering to the cell or expressing in the cell an antisense oligonucleotide comprising a nucleic acid sequence complementary to or capable of hybridizing under stringent conditions to a nucleic acid of the invention. The invention provides an antisense oligonucleotide comprising a nucleic acid sequence complementary to or capable of hybridizing under stringent conditions to a nucleic acid of the invention. The invention provides methods of inhibiting the translation of a phospholipase message in a cell comprising administering to the cell or expressing in the cell an antisense oligonucleotide comprising a nucleic acid sequence complementary to or capable of hybridizing under stringent conditions to a nucleic acid of the invention. . The antisense oligonucleotide can be between about 10 to 50, about 20 to 60, about 30 to 70, about 40 to 80, about 60 to 100, about 70 to 110, or about 80 to 120 length bases. The invention provides methods of inhibiting the translation of a phospholipase, for example a phospholipase message in a cell comprising administering to the cell or expressing in the cell an antisense oligonucleotide comprising a nucleic acid sequence complementary to or capable of hybridizing under stringent conditions. for a nucleic acid of the invention. The invention provides strand inhibitor RNA molecules double (RNAi) comprising a subsequence of a sequence of the invention. In one aspect, the RNAi is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more nucleotide duplexes in length. The invention provides methods of inhibiting the expression of a phospholipase, for example a phospholipase, in a cell comprising administering to the cell or expressing in the cell a double-stranded inhibitory RNA (iRNA), wherein the RNA comprises a subsequence of a sequence of the invention. The invention provides a recombinant or isolated polypeptide comprising an amino acid sequence having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60% , 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or more, or full sequence identity (100%) for an exemplary polypeptide or peptide of the invention (e.g., SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10 , SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO : 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60 , SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO : 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120 or SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO : 172, or SEQ ID NO: 174) over a region of at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 400, 450, 500, 550 or 600 or more residues, or about the natural size of the polypeptide; and, in one aspect, sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. In one aspect, the invention provides a recombinant or isolated polypeptide comprising an amino acid sequence having at least about 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89% , 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for SEQ ID NO: 2. In one aspect, the invention provides a recombinant or isolated polypeptide comprising an amino acid sequence having at least about 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for SEQ ID NO: 4. In one aspect, the invention provides a recombinant or isolated polypeptide comprising an amino acid sequence having at least about 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or identity sequence (100%) for SEQ ID NO: 6. In one aspect, the invention provides a polypeptide recombinant or isolated id comprising an amino acid sequence having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater, or full sequence identity (100%) for SEQ ID NO: 8. The invention provides recombinant or isolated polypeptides encoded by a nucleic acid of the invention. In alternative aspects, the polypeptide may have a sequence mentioned in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO : 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48 , SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO : 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98 , SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 1 26, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO: 174. The polypeptide may have phospholipase activity, for example a phospholipase A, B, C or D activity, or any combination of phospholipase activity, for example a PL A, PL C and / or PL D activity as a multifunctional activity. For example, in one aspect, a polypeptide of the invention is enzymatically active, but requires lipase activity, e.g., requires any enzymatic activity that affects a neutral oil (triglyceride) fraction. In one aspect, the invention provides a desizing process comprising use of a polypeptide of the invention having a phospholipase activity, but not a lipase activity, such that in the desizing process any neutral oil fraction is not impaired (decreased , altered, degraded, e.g. hydrolyzed). The invention provides recombinant or isolated polypeptides comprising a polypeptide of the invention requiring a signal sequence. In one aspect, the polypeptide requiring a signal sequence is at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity for residues 30 to 287 of SEQ ID NO: 2, an amino acid sequence having at least 78%, 79% , 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98%, 99%, or more sequence identity for residues 25 to 283 of SEQ ID NO: 4, an amino acid sequence having at least 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , or more sequence identity to residues 26 to 280 of SEQ ID NO: 6, or, an amino acid sequence having at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57% , 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identities for residues 40 to 330 of SEQ ID NO: 8. Sequence identities can be determined by analysis with a sequence comparison algorithm or by visual inspection. Another aspect of the invention provides an isolated or recombinant polypeptide or peptide including at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 , 95 or 100 or more consecutive bases of a polypeptide or peptide sequence of the invention, sequences substantially identical thereto, and the sequences complementary thereto. The peptide can be, for example, an immunogenic fragment, a motif (for example, a binding site) or an active site. In one aspect, the recombinant or isolated polypeptide of the invention (with or without a signal sequence) has a phospholipase activity. In one aspect, the phospholipase activity comprises catalyzing hydrolysis of a glycerolphosphate ester bond (i.e., cleavage of glycerolphosphate ester bonds). Phospholipase activity can comprise catalyze hydrolysis of an ester bond on a phospholipid in a vegetable oil. The vegetable oil phospholipid may comprise an oilseed phospholipid. The phospholipase activity may comprise a phospholipase C (PLC) activity; a phospholipase A (PLA) activity, such as a phospholipase A1 or phospholipase A2 activity; a phospholipase D (PLD) activity, such as a phospholipase D1 activity or a phospholipase D2 activity; a phospholipase B (PLB) activity, for example, a phospholipase and a lysophospholipase (LPL) activity or a phospholipase and a lysophospholipase-transacylase (LPTA) activity or a phospholipase and a lysophospholipase (LPL) activity and lysophospholipase-transacylase (LPTA) activity; or patatin activity, or a combination thereof. For example, in one aspect a phospholipase comprises a combination of one or more of the enzyme activities described herein, for example, a phospholipase may have PLC and PLA activity; PLB and PLA activity; PLC and PLD activity; PLC and PLB activity; PLB and patatin activity; PLC and patatin activity; PLD and PLA activity; PLD, PLA, PLB and PLC activity; or PLD, PLA, PLB, PLC and patatin activity; or, a phospholipase and a lysophospholipase (LPL) activity or a phospholipase and a lysophospholipase-transacylase (LPTA) activity or a phospholipase and a lysophospholipase (LPL) activity and a lysophospholipase-transacylase (LPTA) activity, or any combination of these. Phospholipase activity may comprise hydrolysis of a glycoprotein, for example as a glycoprotein found in a potato tuber. The phospholipase activity may comprise a patatin enzymatic activity. Phospholipase activity may comprise a lipid acyl hydrolase (LAH) activity. In one aspect, phospholipase activity is thermostable. The polypeptide can maintain a phospholipase activity under conditions comprising a temperature range of from about 20 to about 30 °C, from about 25 °C to about 40 °C, from about 37 °C to about 30 °C. 95°C, between about 55°C to about 85°C, between about 70°C to about 95°C, or between about 90°C to about 95°C. In another aspect, the activity of phospholipase may be thermotolerant. The polypeptide can maintain phospholipase activity after exposure to a temperature in the range greater than 37 °C to about 95 °C, or in the range greater than 55 °C to about 85 °C. In one aspect, the polypeptide can maintain phospholipase activity after exposure to a temperature in the range greater than 90°C to about 95°C at pH 4.5. In one aspect, the polypeptide can maintain a phospholipase activity under conditions comprising about pH 6.5, pH 6, pH 5.5, pH 5, pH 4.5, or pH 4 or less (more acidic). In one aspect, the polypeptide can maintain a phospholipase activity under conditions comprising about pH 7, pH 7.5, pH 8.0, pH 8.5, pH 9, pH 9.5, pH 10, pH 10.5 or pH 11 or more (most basic). In one aspect, the recombinant or isolated polypeptide may comprise the polypeptide of the invention requiring a signal sequence. In one aspect, the recombinant or isolated polypeptide may comprise the polypeptide of the invention comprising a heterologous signal sequence, such as a non-phospholipase or heterologous phospholipase signal sequence. The invention provides recombinant or isolated peptides comprising an amino acid sequence having at least 95%, 96%, 97%, 98%, 99%, or more sequence identity to residues 1 to 29 of SEQ ID NO: 2, at least 95 %, 96%, 97%, 98%, 99%, or more sequence identity for residues 1 to 24 of SEQ ID NO: 4, at least 95%, 96%, 97%, 98%, 99%, or more sequence identity for residues 1 to 25 of SEQ ID NO: 6, or at least 95%, 96%, 97%, 98%, 99%, or more sequence identity for residues 1 to 39 of SEQ ID NO: 8, and for other signal sequences as mentioned in the SEQ ID listing, wherein sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. These peptides can act as signal sequences on their endogenous phospholipase, on another phospholipase, or on a heterologous protein (a non-phospholipase enzyme or another protein). In one aspect, the invention provides chimeric proteins comprising a first domain comprising a signal sequence of the invention and at least one second domain. The protein may be a fusion protein. The second domain may comprise an enzyme. The enzyme may be a phospholipase. The invention provides chimeric polypeptides comprising at least one first domain comprising a signal peptide (SP) of the invention or a catalytic domain (CD), or active site, of a phospholipase of the invention and at least one second domain comprising a polypeptide or peptide. heterologous, wherein the heterologous polypeptide or peptide is not naturally associated with the signal peptide (SP) or catalytic domain (CD). In one aspect, the heterologous polypeptide or peptide is not a phospholipase. The heterologous polypeptide or peptide may be amino-terminal to, carboxy-terminal to, or at both ends of the signal peptide (SP) or catalytic domain (CD). The invention provides recombinant or isolated nucleic acids encoding a chimeric polypeptide, wherein the chimeric polypeptide comprises at least a first domain comprising signal peptide (SP) or a catalytic domain (CD), or active site, of a polypeptide of the invention, and at least a second domain comprising a heterologous polypeptide or peptide, wherein the heterologous polypeptide or peptide is not naturally associated with the signal peptide (SP) or catalytic domain (CD). In one aspect, phospholipase activity comprises a specific activity at about 37°C in the range of about 10 units per milligram to about 100 units per milligram of protein. In another aspect, phospholipase activity comprises a specific activity of about 100 units per milligram to about 1000 units per milligram, from about 500 units per milligram to about 750 units per milligram of protein. Alternatively, phospholipase activity comprises a specific activity at 37°C in the range of about 100 to about 500 units per milligram of protein. In one aspect, the phospholipase activity comprises a specific activity at 37°C in the range of about 500 to about 1200 units per milligram of protein. In another aspect, phospholipase activity comprises a specific activity at 37°C in the range of about 750 to about 1000 units per milligram of protein. In another aspect, thermotolerance comprises retaining at least half of the phospholipase specific activity at 37°C after being heated to an elevated temperature. Alternatively, thermotolerance may comprise retention of specific activity at 37°C in the range of about 500 to about 1200 units per milligram of protein after being heated to an elevated temperature. The invention provides a recombinant or isolated polypeptide of the invention, wherein the polypeptide comprises at least one glycosylation site. In one aspect, the glycosylation may be an N-linked glycosylation. In one aspect, the polypeptide may be glycosylated after being expressed in a P. pastoris or an S. pombe. The invention provides phospholipase enzymes and the nucleic acids encoding them having a sequence of any of the exemplary phospholipases of the invention with one or more or all altered glycosylation sites as described above. Thus, the invention provides methods of preparing variant phospholipase encoding sequences having increased expression in a host cell, wherein the method comprises modifying a phospholipase coding sequence of the invention such that one, several or all of the glycosylation site coding motifs are linked together. by N are modified into a non-glycosylated motif. The invention likewise provides phospholipase coding sequence made by this process, and the enzymes they encode. The invention provides methods for preparing a variant phospholipase coding sequence encoding a phospholipase having increased resistance to a protease comprising modifying an amino acid equivalent to position 131 of SEQ ID NO: 2 into one, several or all of the following residues: Lysine (K) ; Serine (S); Glycine (G); Arginine (R); Glutamine (Q); Alanine (A); Isoleucine (I); Histidine (H); Phenylalanine (F); Threonine (T); Methionine (M) Leucine (L), including variants for SEQ ID NO: 2 (and the acid nucleic acid encoding them) having these exemplary modifications. The invention likewise provides isolated recombinant or synthetic phospholipases encoded by a sequence made by this method. The invention provides methods for preparing a variant phospholipase coding sequence encoding a phospholipase having decreased resistance in a protease comprising modifying an amino acid equivalent to position 131 of SEQ ID NO: 2 into one, several or all of the following residues: Tryptophan (W) ; Glutamate (E); Tyrosine (Y), including variants for SEQ ID NO: 2 (and the nucleic acid encoding them) having these exemplary modifications. The invention likewise provides isolated recombinant or synthetic phospholipases encoded by a sequence made by this method. The invention provides protein preparations comprising a polypeptide of the invention, wherein the protein preparation comprises a liquid, a solid or a gel. The invention provides heterodimers comprising a polypeptide of the invention and a second protein or domain. The second member of the heterodimer can be a different phospholipase, a different enzyme, or another protein. In one aspect, the second domain can be a polypeptide and the heterodimer can be a fusion protein. In one aspect, the second domain may be an epitope or a label. In one aspect, the invention provides homodimers comprising a polypeptide of the invention. The invention provides immobilized polypeptides having a phospholipase activity, wherein the polypeptide comprises a polypeptide of the invention, a polypeptide encoded by a nucleic acid of the invention, or a polypeptide comprising a polypeptide of the invention and a second domain (e.g., a protein of Fusion). In one aspect, a polypeptide of the invention is immobilized on a cell, a vesicle, a liposome, a film, a membrane, a metal, a resin, a polymer, a ceramic, a glass, a microelectrode, a graphite particle, a bead, a gel, a plate, crystals, a tablet, a pill, a capsule, a powder, an agglomerate, a surface, a porous structure, an arrangement or a capillary tube. In one aspect, a polypeptide of the invention is immobilized on materials such as grain, dried husk, rind, skin, hair, enamel, bone, shell and materials derived therefrom, or animal food materials or a combination thereof. Polypeptides of the invention (e.g. phospholipases) may likewise be present alone or as a mixture of phospholipases or phospholipases and other hydrolytic enzymes such as cellulases, xylanases, proteases, lipases, amylases or redox enzymes such as laccases, peroxidases, catalases, oxidases or reductases. They may be formulated in a solid form such as a powder, lyophilized preparations, granules, tablets, bars, crystals, capsules, pills, pellets, or in a liquid form such as an aqueous solution, an aerosol, a gel, a paste, a suspension, an aqueous / oil emulsion, a cream, a capsule, micellar or vesicular suspension. In one aspect, these formulations of the invention may comprise any or a combination of the following ingredients: polyols such as polyethylene glycols, polyvinylalcohols, glycerol, sugars such as sucrose, sorbitol, trehalose, glucose, fructose, maltose, gelling agents such as guar gum, carrageenans, alginates, dextrans, cellulosic derivatives, pectins, salts such as sodium chloride, sodium sulfate, ammonium sulfate, calcium chloride, magnesium chloride, zinc chloride, zinc sulfate, fatty acid salts and derivatives thereof, metal chelators such as EDTA, EGTA, sodium citrate, antimicrobial agents such as fatty acids, derivatives thereof, parabens, sorbates, benzoates, additionally protease impact blocking compounds such as bulk proteins such as BSA, hydrolysates wheat, borate compounds, emulsifiers such as non-ionic and ionic detergents can be used alone or in combination, phytosterols, vitamins, amino acids, reducing agents such as cysteine ​​compounds or an antioxidant such as ascorbic acid may be included as well as dispersants. In one aspect, cross-linking and protein modification such as pegylation, fatty acid modification, and glycosylation are employed to improve the stability of a polypeptide of the invention (e.g., this enzyme ability). In one aspect, the polyols and / or sugars comprise from about 5% to about 60% or more of the formulation, from about 10% to about 50% of the formulation, from about 20% to about 40 % of the formulation, or from about 5% to about 20% of the formulation. In another aspect, The gelling agents comprise from about 0.5% to about 10% of the formulation, from about 1% to about 8% of the formulation, from about 2% to about 5% of the formulation, or from about from 0.5% to about 3% of the formulation. In another aspect, salts such as sodium chloride, sodium sulfate, ammonium sulfate, calcium chloride and / or magnesium chloride comprise from about 1% to about 30% of the formulation, from about 2% to about 20% of the formulation, from about 5% to about 15% of the formulation, or from about 1% to about 10% of the formulation. In another aspect, zinc chloride is present in the formulation in concentrations comprising from about 0.1 mM to about 20 mM, from about 0.5 mM to about 10 mM, from about 1 mM to about 5 mM. mM, or from about 0.1 mM to about 5 mM). In yet another aspect, zinc sulfate is present in the formulation at concentrations comprising from about 0.1 mM to about 20 mM, from about 0.5 mM to about 10 mM, from about 1 mM to about 5 mM. mM, or from about 0.1 mM to about 5 mM). In another aspect, fatty acid salts and / or derivatives thereof comprise from about 5% to about 40% of the formulation, from about 10% to about 30% of the formulation, from about 15% to about 25% of the formulation. % of the formulation, or from about 5% to about 20% of the formulation. In another aspect, metal chelators such as EDTA, EGTA, and / or sodium citrate are present in the formulation at concentrations ranging from 0.1 mM to about 10 mM), from about 0.5 mM to about 0.5 mM to about 10 mM. of 8 mM, from about 1 mM to about 5 mM, or from about 0.1 mM to about 1 mm In another aspect, antimicrobials such as parabens, sorbates and / or benzoates comprise from about 0.01% to about 10% of the formulation, from about 0.05% to about 5% of the formulation, from about 0. 1% to about 1% of the formulation, or from about 0.05% to about 0.5% of the formulation. In yet another aspect, bulk proteins such as BSA and / or wheat hydrolysates comprise from about 1% to about 20% of the form formulation, from about 5% to about 15% of the formulation, from about 2.5% to about 7.5% of the formulation, or from about 1% to about 5% of the formulation. ' lation. In another aspect, emulsifiers such as ionic and / or non-ionic detergents are present in the formulation in concentrations comprising 5 ranging from about 1X critical micelle (CMC) concentration to about 10X CMC, from about 2.5X CMC to about 7.5X CMC, from about 1X CMC to about 5X CMC , or from about 3X CMC to about 6X CMC. In another aspect, vitamins, amino acids, reducing agents and / or antioxidant compounds comprise from about 0.1% to about 5% of the formulation, from about 0.5% to about 4% of the formulation, from about 0.5% to about 4% of the formulation. from 1% to about 2.5% of the formulation, or from about 0.1% to about 1% of the formulation. The invention provides provisions comprising an immobilized polypeptide, wherein the polypeptide is a phospholipase of the invention or is a polypeptide encoded by a nucleic acid of the invention. The invention provides arrangements comprising an immobilized nucleic acid of the invention. The invention provides an arrangement comprising an immobilized antibody of the invention. The invention provides recombinant or isolated antibodies that specifically bind to a polypeptide of the invention or a polypeptide encoded by a nucleic acid of the invention. The antibody may be a monoclonal or a polyclonal antibody. The invention provides hybridomas comprising an antibody of the invention. The invention provides methods of isolating or identifying a polypeptide with a phospholipase activity comprising the steps of: (a) providing an antibody of the invention; (b) providing a sample comprising polypeptides; and, (c) contacting the sample from step (b) with the antibody from step (a) under conditions where the antibody can specifically bind the polypeptide, thereby isolating or identifying a phospholipase. The invention provides methods of preparing an antiphospholipase antibody comprising administering to a non-human animal a nucleic acid of the invention, or a polypeptide of the invention, in an amount sufficient to generate a humoral immune response, thereby preparing an antiphospholipase antibody. The invention provides methods of producing a recombinant polypeptide comprising the steps of: (a) providing a nucleic acid of the invention operably linked to a promoter; and, (b) expressing the nucleic acid of step (a) under conditions that allow expression of the polypeptide, thereby producing a recombinant polypeptide. The nucleic acid may comprise a sequence having at least 85% sequence identity to SEQ ID NO: 1 over a region of at least about 100 residues, having at least 80% sequence identity to SEQ ID NO: 3 over a region of at least about 100 residues. region of at least about 100 residues, having at least 80% sequence identity to SEQ ID NO: 5 over a region of at least about 100 residues, or having at least 70% sequence identity to SEQ ID NO: 7 over a region of at least about 100 residues, where sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. The nucleic acid may comprise a nucleic acid that hybridizes under stringent conditions to a nucleic acid as mentioned in SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; or, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof. The method may also comprise transforming a host cell with the nucleic acid of step (a) followed by expressing the nucleic acid of step (a), thereby producing a recombinant polypeptide in a transformed cell. The method may also comprise inserting the nucleic acid of step (a) into a non-human host animal followed by expressing the nucleic acid of step (a), thereby producing a recombinant polypeptide in the non-human host animal. The invention provides methods for identifying a polypeptide having a phospholipase activity comprising the following steps: (a) providing a polypeptide of the invention or a polypeptide encoded by a nucleic acid of the invention or a fragment or variant thereof, (b) providing a phospholipase substrate; and, (c) contacting the polypeptide or a fragment or variant thereof from step (a) with the substrate from step (b) and detecting an increase in the amount of substrate or a decrease in the amount of reaction product, wherein a decrease in the amount of substrate or an increase in the amount of reaction product detects a polypeptide having a phospholipase activity. In alternative aspects, the nucleic acid comprises a sequence having at least 85% sequence identity to SEQ ID NO: 1 over a region of at least about 100 residues, having at least 80% sequence identity to SEQ ID NO: 3 over a region of at least about 100 residues, having at least 80% sequence identity to SEQ ID NO: 5 over a region of at least about 100 residues, or having at least 70% sequence identity to SEQ ID NO: 7 over a region of at least about 100 residues, wherein sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. In alternative aspects the nucleic acid hybridizes under stringent conditions to a sequence as mentioned in SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; or, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof. The invention provides methods for identifying a phospholipase substrate comprising the following steps: (a) providing a polypeptide of the invention or a polypeptide encoded by a nucleic acid of the invention; (b) providing a test substrate; and, (c) contacting the polypeptide of step (a) with the test substrate of step (b) and detecting an increase in the amount of substrate or a decrease in the amount of reaction product, wherein a decrease in the amount of substrate or a increase in the amount of reaction product identifies the test substrate as a phospholipase substrate. In alternative aspects, the nucleic acid may have at least 85% sequence identity to SEQ ID NO: 1 over a region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 3 over a region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 5 over a region of at least about 100 residues, or, at least 70% sequence identity for SEQ ID NO: 7 over a region of at least about 100 residues, wherein sequence identities are determined by analysis with a matching algorithm. sequence comparison or by visual inspection. In alternative aspects, the nucleic acid hybridizes under stringent conditions to a sequence as mentioned in SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; or, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof. The invention provides methods of determining whether a compound specifically binds a phospholipase comprising the following steps: (a) expressing a nucleic acid or a vector comprising the nucleic acid under permissive conditions for translation of the nucleic acid into a polypeptide, wherein the nucleic acid and vector comprise a nucleic acid or vector of the invention; or, providing a polypeptide of the invention (b) contacting the polypeptide with the test compound; and, (c) determining whether the test compound specifically binds the polypeptide, thereby determining that the compound specifically binds phospholipase. In alternative aspects, the nucleic acid sequence has at least 85% sequence identity to SEQ ID NO: 1 over a region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 3 over a region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 5 over a region of at least about 100 residues, or, at least 70% sequence identity to SEQ ID NO: 7 over a region of at least about 100 residues, where sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. In alternative aspects, the nucleic acid hybridizes under rigorous conditions. roses for a sequence as mentioned in SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; or, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof. The invention provides methods for identifying a modulator of a phospholipase activity comprising the following steps: (a) providing a polypeptide of the invention or a polypeptide encoded by a nucleic acid of the invention; (b) providing a test compound; (c) contacting the step (a) polypeptide with the step (b) test compound; and, measuring a phospholipase activity, wherein a change in phospholipase activity measured in the presence of test compound compared to activity in the absence of test compound provides a determination that the test compound modulates phospholipase activity. In alternative aspects, the nucleic acid may have at least 85% sequence identity to SEQ ID NO: 1 over a region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 3 over a region of at least about 100 residues. region of at least about 100 residues, at least 80% sequence identity to SEQ ID NO: 5 over a region of at least about 100 residues, or, at least 70% sequence identity to SEQ ID NO: 7 over a region of at least about 100 residues, where sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection. In alternative aspects, the nucleic acid may hybridize under stringent conditions to a nucleic acid sequence selected from the group consisting of a sequence as mentioned in SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; and, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof. In one aspect, phospholipase activity is measured by providing a phospholipase substrate and detecting an increase in the amount of the substrate or a decrease in the amount of a reaction product. The decrease in the amount of substrate or the increase in the amount of reaction product with the test compound when compared to the amount of substrate or reaction product without the test compound identifies the test compound as an activator of phospholipase activity. The increase in the amount of substrate or the decrease in the amount of reaction product with the test compound when compared to the amount of substrate or reaction product without the test compound identifies the test compound as an inhibitor of phospholipase activity. The invention provides computer systems comprising a processor and a data storage device wherein said data storage device has stored therein a polypeptide sequence of the invention or a nucleic acid sequence of the invention. In one aspect, the computer system may also comprise a sequence comparison algorithm and a data storage device having at least one reference sequence stored therein. The sequence comparison algorithm may comprise a computer program that indicates polymorphisms. The computer system may also comprise an identifier that identifies one or more features in said sequence. The invention provides computer readable media having stored therein a sequence comprising a polypeptide sequence of the invention or a nucleic acid sequence of the invention. The invention provides methods for identifying a feature in a sequence comprising the steps of: (a) reading the sequence employing a computer program that identifies one or more features in a sequence, wherein the sequence comprises a polypeptide sequence of the invention or a nucleic acid sequence of the invention; and, (b) identifying one or more features in the sequence with the computer program. The invention provides methods for comparing a first sequence to a second sequence comprising the steps of: (a) reading the first sequence and the second sequence through the use of a sequence comparing computer program, wherein the first sequence comprises a polypeptide sequence of the invention or an acid sequence nucleic of the invention; and, (b) determining differences between the first sequence and the second sequence with the computer program. In one aspect, the step of determining differences between the first sequence and the second sequence also comprises the step of identifying polymorphisms. In one aspect, the method also comprises an identifier (and use of the identifier) ​​that identifies one or more features in a sequence. In one aspect, the method comprises reading the first sequence using a computer program and identifying one or more features in the sequence. The invention provides methods for isolating or recovering a nucleic acid encoding a polypeptide with a phospholipase activity from an environmental sample comprising the steps of: (a) providing an amplification primer sequence pair to amplify a nucleic acid encoding a polypeptide having a phospholipase activity, wherein the primer pair is capable of amplifying a nucleic acid of the invention (e.g., SEQ ID NO: 1, or a subsequence thereof; SEQ ID NO: 3, or a subsequence thereof; SEQ ID NO: : 5, or a subsequence thereof; or SEQ ID NO: 7, or a subsequence thereof, etc.); (b) isolating a nucleic acid from the environmental sample or treating the environmental sample such that the nucleic acid in the sample is accessible for hybridization to the amplification primer pair; and, (c) combining the nucleic acid from step (b) with the amplification primer pair from step (a) and amplifying the nucleic acid from the environmental sample, thereby isolating or recovering a nucleic acid encoding a polypeptide with a phospholipase activity from an environmental sample. In one aspect, each amplification sequence primer pair member comprises an oligonucleotide comprising at least about 10 to 50 consecutive bases of a nucleic acid sequence of the invention. In one aspect, the sequence initiator pair amplification sequence is an amplification pair of the invention. The invention provides methods for isolating or recovering a nucleic acid encoding a polypeptide with a phospholipase activity from an environmental sample comprising the steps of: (a) providing a polynucleotide probe comprising a nucleic acid sequence of the invention, or a subsequence of this; (b) isolating a nucleic acid from the environmental sample or treating the environmental sample such that the nucleic acid in the sample is accessible for hybridization to a polynucleotide probe of step (a); (c) combining the isolated nucleic acid or treated environmental sample from step (b) with the polynucleotide probe from step (a); and, (d) isolating a nucleic acid that specifically hybridizes to the polynucleotide probe of step (a), thereby isolating or recovering a nucleic acid encoding a polypeptide with a phospholipase activity from the environmental sample. In alternative aspects, the environmental sample comprises a water sample, a liquid sample, a soil sample, an air sample, or a biological sample. In alternative aspects, the biological sample is derived from a bacterial cell, a protozoan cell, an insect cell, a yeast cell, a plant cell, a fungal cell, an algal cell (algae), a lichen, or a mammalian cell. The invention provides methods of generating a nucleic acid variant encoding a phospholipase comprising the steps of: (a) providing a standard nucleic acid comprising a nucleic acid of the invention; (b) modifying, deleting or adding one or more nucleotides in the standard sequence, or a combination thereof, to generate a variant of the standard nucleic acid. In one aspect, the method also comprises expressing the variant nucleic acid to generate a variant phospholipase polypeptide. In alternative aspects, the modifications, additions or deletions are introduced by error-prone PCR, scrambling, oligonucleotide-directed mutagenesis, assembly PCR, sexual PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive set mutagenesis, exponential ensemble mutagenesis, site-specific mutagenesis, gene reassembly, Gene Site Saturation Mutagenesis® (GSSM®), synthetic linkage reassortment (SLR) and / or a combination thereof. In alternative aspects, the modifications, additions or detections are introduced by a method selected from the group consisting of recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, standard uracil-containing mutagenesis, duplex-spaced mutagenesis, point imperfect linkage repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiogenic mutagenesis, deletion mutagenesis, restriction-selection mutagenesis, restriction-purification mutagenesis, artificial gene synthesis, ensemble mutagenesis, multimer creation of chimeric nucleic acid and / or a combination thereof. In one aspect, the method is iteratively repeated until a phospholipase having a different or altered activity or different stability than a standard nucleic acid-encoded phospholipase is produced. In one aspect, the different or altered activity is a phospholipase activity under an acidic condition, wherein the phospholipase encoded by the standard nucleic acid is not active under the acidic condition. In one aspect, different or altered activity is a phospholipase activity under high temperature, where the phospholipase encoded by the standard nucleic acid is not active under high temperature. In one aspect, the method is iteratively repeated until a phospholipase coding sequence having a codon usage changed from that of the standard nucleic acid is produced. The method can be iteratively repeated until a phospholipase gene having a higher or lower level of message expression or stability than the standard nucleic acid is produced. The invention provides methods for modifying codons in a nucleic acid encoding a phospholipase to increase its expression in a host cell, the method comprising (a) providing a nucleic acid of the invention encoding a phospholipase; and, (b) identifying a less-preferred or non-preferred codon in the nucleic acid of step (a) and replacing it with a neutrally employed or preferred codon encoding the same amino acid as the substituted codon, wherein a preferred codon is an over-represented codon encoding sequences in genes in the host cell and a less-preferred or non-preferred codon is an under-represented codon encoding sequences in genes in the host cell, thereby modifying the nucleic acid to increase its expression in a host cell. The invention provides methods for modifying codons in a nucleic acid encoding a phospholipase, the method comprising (a) providing a nucleic acid of the invention encoding a phospholipase; and, (b) identifying a codon in the nucleic acid of step (a) and replacing it with a different codon encoding the same amino acid as the substituted codon, thereby modifying codons in a nucleic acid encoding a phospholipase. The invention provides methods for modifying codons in a nucleic acid encoding a phospholipase to increase its expression in a host cell, the method comprising (a) providing a nucleic acid of the invention encoding a phospholipase; and, (b) identifying a less preferred or non-preferred codon in the nucleic acid of step (a) and replacing it with a neutrally preferred or preferred codon encoding the same amino acid as the substituted codon, wherein a preferred codon is a codon above -represented in coding sequences in genes in the host cell and a less preferred or non-preferred codon is a codon underrepresented in coding sequences in genes in the host cell, thereby modifying the nucleic acid to increase its expression in a host cell. The invention provides methods for modifying a codon in a nucleic acid encoding a phospholipase to decrease its expression in a host cell, the method comprising (a) providing a nucleic acid of the invention encoding a phospholipase; and, (b) identifying at least one preferred codon in the nucleic acid of step (a) and replacing it with a less preferred or non-preferred codon encoding the same co-amino acid. such as the substituted codon, wherein a preferred codon is an overrepresented codon in gene coding sequences in a host cell and a less preferred or non-preferred codon is an underrepresented codon in gene coding sequences in the host cell, thereby modifying the nucleic acid to decrease its expression in a host cell. In alternative aspects, the host cell is a bacterial cell, a fungal cell, an insect cell, a yeast cell, a plant cell, an alga (algae) cell, a lichen, or a mammalian cell. The invention provides methods for producing a library of nucleic acids encoding a plurality of modified phospholipase active sites or substrate binding sites, wherein the modified active sites or substrate binding sites are derived from a first nucleic acid comprising a sequence encoding a first active site or a first substrate binding site, the method comprising: (a) providing a first nucleic acid encoding a first active site or first substrate binding site, wherein the first nucleic acid sequence comprises a nucleic acid of the invention; (b) providing a set of mutagenic oligonucleotides that encode naturally occurring amino acid variants at a plurality of codons targeted at the first nucleic acid; and, (c) employing the set of mutagenic oligonucleotides to generate a set of substrate-binding-site-encoding or active-site-encoding variant nucleic acids encoding a range of amino acid variations at each amino acid codon that has been mutagenized, thereby producing a nucleic acid library encoding a plurality of modified phospholipase active sites or substrate binding sites. In alternative aspects, the method comprises mutagenizing the first nucleic acid of step (a) by a method comprising an optimized targeted evolution system, Gene Site Saturation Mutagenesis® (GSSM®), and synthetic linkage reassembly (SLR). The method may also comprise mutagenizing the first nucleic acid of step (a) or variants by a method comprising PCR prone to error, scrambling, oligonucleotide-directed mutagenesis, pool PCR, sex PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive set mutagenesis, exponential set mutagenesis, site-specific mutagenesis, gene reassembly, Gene Site Saturation Mutagenesis® ( GSSM®), synthetic bond reassembly (SLR) and a combination thereof. The method may also comprise mutagenizing the first nucleic acid of step (a) or variants by a method comprising recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, standard uracil-containing mutagenesis, gap-duplex mutagenesis, point imperfect linkage repair, repair-deficient host strain mutagenesis, chemical mutagenesis, radiogenic mutagenesis, deletion mutagenesis, restriction-selection mutagenesis, restriction-purification mutagenesis, artificial gene synthesis, ensemble mutagenesis, acid multimer creation chimeric nucleic acid and a combination thereof. The invention provides methods for preparing a small molecule comprising the steps of: (a) providing a plurality of biosynthetic enzymes capable of synthesizing or modifying a small molecule, wherein one of the enzymes comprises a phospholipase enzyme encoded by a nucleic acid of the invention; (b) providing a substrate for at least one of the enzymes of step (a); and, (c) reacting the substrate from step (b) with the enzymes under conditions that facilitate a plurality of biocatalytic reactions to generate a small molecule by a series of biocatalytic reactions. The invention provides methods for modifying a small molecule comprising the steps: (a) providing a phospholipase enzyme encoded by a nucleic acid of the invention; (b) providing a small molecule; and, (c) reacting the enzyme from step (a) with the small molecule from step (b) under conditions that facilitate an enzymatic reaction catalyzed by the phospholipase enzyme, thereby modifying a small molecule by an enzymatic phospholipase reaction. In one aspect, the method comprises providing a plurality of small molecule substrates for the enzyme of step (a), thereby generating a library of modified small molecules produced by at least one enzymatic reaction catalyzed by the phospholipase enzyme. In one aspect, the method also comprises a plurality of additional enzymes under conditions that facilitate a plurality of biocatalytic reactions by the enzymes to form a library of modified small molecules produced by the plurality of enzymatic reactions. In one aspect, the method also comprises the step of testing the library to determine whether a particular modified small molecule that exhibits a desired activity is present within the library. The step of testing the library may also comprise the steps of systemically eliminating all but one of the biocatalytic reactions employed to produce a portion of the plurality of modified small molecules within the library by testing the modified small molecule portion for the presence or absence of the molecule. particular modified small molecule with a desired activity, and identifying at least one specific biocatalytic reaction that produces the particular modified small molecule of the desired activity. The invention provides methods for determining a functional fragment of a phospholipase enzyme comprising the steps of: (a) providing a phospholipase enzyme comprising an amino acid sequence of the invention; and, (b) deleting a plurality of amino acid residues from the sequence of step (a) and testing the remaining subsequence for phospholipase activity, thereby determining a functional fragment of a phospholipase enzyme. In one aspect, phospholipase activity is measured by providing a phospholipase substrate and detecting an increase in the amount of substrate or a decrease in the amount of a reaction product. In one aspect, a decrease in the amount of an enzyme substrate or an increase in the amount of reaction product with the test compound as compared to the amount of substrate or reaction product without the test compound identifies the test compound as an activator of enzyme activity. phospholipase. The invention provides methods for cleaving an ester bond from glycerolphosphate comprising the following steps: (a) providing a polypeptide having a phospholipase activity, wherein the polypeptide comprises an amino acid sequence of the invention, or the polypeptide is encoded by a nucleic acid of the invention; (b) providing a composition comprising a glycerolphosphate ester linkage; and, (c) contacting the polypeptide of step (a) with the composition of step (b) under conditions in which the polypeptide cleaves the glycerolphosphate ester bond. In one aspect, the conditions range from about pH 5 to about 8.5, or, from about pH 4.5 (or more acidic, i.e., pH < 4.5) to about 9.0 (or more alkaline ( i.e. pH > 9.) In one aspect, the conditions comprise a temperature of between about 40°C and about 70°C. In one aspect, the composition comprises a vegetable oil. In one aspect, the composition comprises a phosphor -oilseed lipid In one aspect, the cleavage reaction can generate a water-extractable phosphorylated base and a diglyceride. The invention provides methods of hydrolyzing, separating or disrupting a phospholipid comprising composition comprising providing at least one polypeptide of the invention having a phospholipase activity or a polypeptide having a phospholipase activity encoded by at least one nucleic acid of the invention; providing a composition comprising a phospholipid; and contacting the polypeptide with the composition under conditions in which the phospholipase hydrolyzes, separates or disrupts the composition comprising phospholipid. In one aspect, the method comprises using high shear mixing of the composition, followed by none or a low shear mixing with the at least one polypeptide of the invention that has a phospholipase activity to allow adequate "contact" of the phospholipid substrate with the phospholipase. The at least one polypeptide that has a phospholipase activity may likewise be present in the high-shear mixing step. The process may be practiced on any scale, for example, on a scale comprising from about 1 gram (g) to about 500, 1000, 2000, 2500, 5000 g, or more, or any amount in this range. The invention provides methods for desizing oil comprising taking the following steps: (a) providing at least one polypeptide that has a phospholipase activity, wherein the polypeptide comprises an amino acid sequence of the invention, or the polypeptide is encoded by a nucleic acid of the invention; (b) providing a composition comprising a vegetable oil; and, (c) contacting the polypeptide from step (a) and the vegetable oil from step (b) under conditions in which the polypeptide can cleave ester bonds in the vegetable oil, thereby degumming the oil. In one aspect, the vegetable oil comprises oilseed. The vegetable oil may comprise rice bran oil, palm oil, rapeseed oil, corn oil, soybean oil, canola oil, sesame oil, peanut oil or sunflower oil. In one aspect, the method also comprises adding a phospholipase of the invention, another phospholipase, or a combination thereof. In one aspect, more than one polypeptide having a phospholipase activity is added to the process, wherein at least one polypeptide is an enzyme of the invention. In one aspect, enzymes are added in a specific order, e.g. PLCs with different specificities are added in a specific order, e.g. an enzyme with PC and PE activity is added first (or two enzymes are added, one with PC and the other with PE activity), then an enzyme with PLC activity of PI is added, or any combination thereof. In one aspect of the oil degumming process, the oil-comprising composition comprises a plant, an animal, an alga, or a fish oil or fat. The plant oil may comprise a rice bran oil, a soybean oil, a rapeseed oil, a corn oil, a palm seed oil, a canola oil, a sunflower oil, a sesame oil or peanut oil. The polypeptide can hydrolyze a phosphatide from a hydratable and / or a non-hydratable phospholipid in the oil-comprising composition. In one aspect, the polypeptide hydrolyzes a phosphatide in a glyceryl phosphoester bond to generate a water-soluble diglyceride and phosphate compound. In one aspect, the polypeptide has a phospholipase C activity. In In one aspect, the polypeptide is a phospholipase D and a phosphatase enzyme is added in the same way. In one aspect of the oil degumming process, contacting comprises hydrolysis of a hydrated phospholipid to an oil. The hydrolysis conditions may comprise alkaline conditions, for example, in one aspect, the conditions comprise a temperature of from about 20°C to 40°C at alkaline pH. Alkaline conditions can comprise a pH of about pH 8 to pH 10, or more. The hydrolysis conditions can be made alkaline at any time in the process, for example, in one aspect, a phospholipase, such as a PLC, is added before the conditions are made alkaline (for example, a "caustic neutralization" of an oil comprising acid, such as phosphatidic acid). In one aspect of the oil degumming process, the base causes the isomerization of 1,2-DAG, produced by PLC, to 1,3-DAG which provides a nutritional health benefit over 1,2-DAG, e.g. 1,3-DAG is burned as energy instead of being stored as fat (as is 1,2-DAG). Thus, the invention provides a process for refining caustic oil in which a phospholipase, e.g. an enzyme of the invention, including a PLC, is added "at the front end", i.e. before adding any acid and caustic, e.g. example, as illustrated in the exemplary process of figure 13. One of the consequences of adding the PLC at the front end of a caustic refinement process of the invention (see also discussion, below), and adding the acid and caustic subsequently, is the generation of a elevated level of 1,3-DAG (not 1,2-DAG). This may be a consequence of base or acid catalyzed acyl migration. Nutritionally, 1,3-DAG is better than 1,2-DAG. Thus, the invention comprises an oil degumming process employing an inventive PLC whereby the final degummed oil product contains no less than about 0.5%, 1.0%, 2.0%, 3 .0%, 4.0% or 5.0% of 1,3-DAG. In one aspect of the oil degumming process, the hydrolysis conditions may comprise a reaction time of about from 3 to 10 minutes or more. The hydrolysis conditions may comprise hydrolysis of non-hydratable and hydratable phospholipids in oil at a temperature between about 50°C to 60°C, at a pH between about pH 5 to pH 6.5, or between about pH 5 at pH 7.5, or between about pH 5 to pH 8.0, employing a reaction time of about 30 to 60 minutes. In one aspect of the oil degumming process, the polypeptide is attached to a filter and the phospholipid-containing oil or fat is passed through the filter. The polypeptide can be added to a solution comprising the phospholipid-containing oil or fat and then the solution is passed through a filter. In one aspect, the oil desizing method also comprises the physical removal of gum produced by the desizing process by the addition of a hardening substance, for example, talc or the like. In one aspect, this increases oil gain. The invention likewise provides methods for converting a non-hydratable phospholipid into a hydratable form comprising the following steps: (a) providing a polypeptide having a phospholipase activity, wherein the polypeptide comprises an amino acid sequence of the invention, or the polypeptide is encoded by a nucleic acid of the invention; (b) providing a composition comprising a non-hydratable phospholipid; and, (c) contacting the polypeptide from step (a) and the non-hydratable phospholipid from step (b) under conditions where the polypeptide can cleave ester bonds in the non-hydratable phospholipid, thereby converting a non-hydratable phospholipid in a hydratable form. The invention provides methods for desizing an oil comprising the following steps: (a) providing a composition comprising a polypeptide of the invention that has a phospholipase activity or a polypeptide encoded by a nucleic acid of the invention; (b) providing a composition comprising a fat or an oil comprising a phospholipid; and (c) contacting the polypeptide of step (a) and the composition of step (b) under conditions under which the polypeptide can degum the phospholipid-comprising composition (under conditions under which the polypeptide can be degummed). deo of the invention can catalyze the hydrolysis of a phospholipid). In one aspect, the oil-comprising composition comprises a plant, an animal, an alga or a fish oil. The plant oil may comprise a rice bran oil, a soybean oil, a rapeseed oil, a corn oil, a palm seed oil, a canola oil, a sunflower oil, a sesame oil or a peanut oil. The polypeptide can hydrolyze a phosphatide from a hydratable and / or a non-hydratable phospholipid in the oil-comprising composition. The polypeptide can hydrolyze a phosphatide in a 10-glyceryl phosphoester bond to generate a water-soluble diglyceride phosphate compound. The polypeptide may have a phospholipase C, B, A or D activity. In one aspect, a phospholipase D activity and a phosphatase enzyme are added. The contact may comprise hydrolysis of a hydrated phospholipid in an oil. The hydrolysis conditions may comprise a temperature of about 20°C to 40°C at an alkaline pH. Alkaline conditions can comprise a pH of about pH 8 to pH 10. Hydrolysis conditions can comprise a reaction time of about 3 to 10 minutes. The hydrolysis conditions may comprise hydrolysis of non-hydratable and hydratable phospholipids in oil at a temperature of about 50°C 20 to 60°C, at a pH of about pH 5 to pH 6.5 employing a reaction time of about 30 to 60 minutes. The polypeptide can be attached to a filter and the phospholipid-containing oil or fat is passed through the filter. The polypeptide can be added to a solution comprising the phospholipid-containing oil or fat and then the solution is passed through a filter. The invention provides methods for converting a non- hydratable in a hydratable form comprising the following steps: (a) providing a composition comprising a polypeptide having a phospholipase activity of the invention, or a polypeptide encoded by a nucleic acid of the invention; (b) providing a composition comprising a non-hydratable phospholipid; and (c) contacting the polypeptide of step (a) and the composition of step (b) under conditions in which the polypeptide converts the non-hydratable phospholipid to a hydratable form. The polypeptide may have a phospholipase C activity. The polypeptide may have a phospholipase D activity and a phosphatase enzyme is likewise added. The invention provides methods for caustic refinement of a phospholipid-containing composition comprising the following steps: (a) providing a composition comprising a phospholipase which may be a polypeptide of the invention that has a phospholipase activity or a polypeptide encoded by a nucleic acid of the invention; (b) providing a composition comprising a phospholipid; and (c) contacting the polypeptide of step (a) with the composition of step (b) before, during or after caustic refinement. The polypeptide may have a phospholipase activity, for example, PLC, PLB, PLD and / or PLA activity. The polypeptide can be added prior to caustic refinement, i.e. at the "front end" of the process, before adding acid or caustic, as illustrated in Figure 13. The polypeptide (which can be an enzyme, eg a PLC, of ​​the invention) can be added during caustic refinement and varying levels of acid and caustic can be added depending on phosphorus levels and free fatty acid levels. The polypeptide (which may be an enzyme of the invention) may be added before caustic refinement, or, after caustic refinement: in an intense mixer or holding mixer before separation; following a heating step; in a centrifuge; into a soap raw material; in a wash water; and / or, during bleaching or deodorizing steps. The method may comprise using concentrated solutions of caustic, for example more concentrated than the industry standard of 11%, to decrease the gum mass. In alternative aspects, the concentrated caustic solution is between about 12% and 50% concentrated, for example, about 20%, 30%, 40%, 50%, or 60%, or more, concentrated. The composition comprising the phospholipid may comprise a plant. The polypeptide can be expressed transgenically in the plant. The polypeptide that has a phospholipase activity can be added during crushing of a seed or other plant part, or, the polypeptide that has a phospholipase activity is added following crushing or before refinement. Also provided is a caustic refinement process for hydrolyzing phospholipids in oil (e.g. plant oil) employing a polypeptide of the invention to generate water-soluble diacylglycerol (DAG) and phosphate ester. In one aspect, the enzyme of the invention must operate in a caustic refinement process, optionally including reduced water and / or in a temperature range of about 55°C to about 70°C. Use of a reduced water caustic refinement process in this temperature range will maximize yield by increasing DAG and reducing captured oil. In one aspect, the enzyme employed in this caustic refinement process of the invention also has very good activity on phosphatidylcholine (PC) and phosphatidylethanolamine (PE), is active between a pH of about pH 6 to pH 9, is active up to 75°C , and is active in water reduced in oil, e.g. about 2% to 5% water, e.g. the enzyme encoded by the sequence of SEQ ID NO: 2, encoded e.g. by SEQ ID NO: 1. In another aspect of the caustic refinement process of the invention for hydrolyzing phospholipids in oils, two enzymes are employed: a PI-specific PLC (hydrolyzes PI), and a PC-PLC which hydrolyzes to PC, PE and PA. This modality generates oil suitable for chemical or physical refinement and maximizes the increase in DAG yield and less oil captured. The invention provides methods for purifying a phytosterol or a triterpene comprising the following steps: (a) providing a composition comprising a polypeptide of the invention which has a phospholipase activity, or a polypeptide encoded by a nucleic acid of the invention; (b) providing a composition comprising a phytosterol or a triterpene; and (c) contacting the polypeptide of step (a) with the composition of step (b) under conditions under which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition. The polypeptide may have a phospholipase C activity. The phytosterol or a triterpene may comprise a plant sterol. Plant sterol can be derived from a vegetable oil. O vegetable oil may comprise rice bran oil, coconut oil, canola oil, cocoa butter oil, corn oil, cottonseed oil, linseed oil, olive oil, palm oil, peanut oil , oil derived from a rice bran, safflower oil, sesame oil, soybean oil or a sunflower oil. The method may comprise using non-polar solvents to quantitatively extract free phytosterols and phytosteryl fatty acid esters. The phytosterol or a triterpene may comprise a p-sitosterol, a campesterol, a stigmasterol, a stigmastanol, a p-sitostanol, a sitostanol, a desmosterol, a calinasterol, a poriferasterol, a clionasterol or a brassicasterol. The invention provides methods for refining a crude oil comprising the following steps: (a) providing a composition comprising a polypeptide of the invention that has a phospholipase activity, or a polypeptide encoded by a nucleic acid of the invention; (b) providing a composition comprising an oil comprising a phospholipid; and (c) contacting the polypeptide of step (a) with the composition of step (b) under conditions under which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition. The polypeptide may have a phospholipase C activity. The polypeptide may have a phospholipase activity that is in a water solution that is added to the composition. The water level can be between about 0.5 to 5%. Process time can be less than about 2 hours, less than about 60 minutes, less than about 30 minutes, less than 15 minutes, or less than 5 minutes. The hydrolysis conditions may comprise a temperature of between about 25°C-70°C. Hydrolysis conditions may comprise use of caustics. Concentrated caustic solutions, for example, more concentrated than the industry standard of 11%, to decrease the gum mass can be employed. In alternative aspects, the concentrated caustic solution is between about 12% and 50% concentrated, for example, about 20%, 30%, 40%, 50%, or 60% or more concentrated. The hydrolysis conditions can comprise a pH between about pH 3 and pH 10, between about pH 4 and pH 9, or between about pH 5 and pH 8. Hydrolysis conditions may comprise addition of emulsifiers and / or mixing after contacting step (c). The methods may comprise adding an emulsion breaker and / or heating or cooling (e.g., from about 4°C to about -20°C, or less) to promote separation of an aqueous phase. The methods may comprise degumming prior to the contacting step to collect lecithin by centrifugation and then adding a PLC, a PLC and / or a PLA to remove non-hydratable phospholipids. The methods may comprise water degumming from crude oil to less than 10 ppm phosphorus for edible oils and subsequent physical refinement to less than about 50 ppm phosphorus for biodiesel oils. The methods may comprise addition of acid to promote hydration of non-hydratable phospholipids. In one aspect, the addition of acid promotes a decrease in the metal content of magnesium and calcium. The invention provides a method of ameliorating or preventing lipopolysaccharide-mediated toxicity comprising administering to a patient a pharmaceutical composition comprising a polypeptide of the invention. The invention provides a method for detoxifying an endotoxin comprising contacting the endotoxin with a polypeptide of the invention. The invention provides a method for deacylating a 2' or a 3' fatty acid chain of a lipid A comprising contacting lipid A with a polypeptide of the invention. The invention provides a method for refining a lubricant comprising the following steps: (a) providing a composition comprising an enzyme of the invention; (b) providing a lubricant; and (c) treating the lubricant with an enzyme under conditions where the enzyme can selectively hydrolyze oils in the lubricant, thereby refining them. The lubricant can be a hydraulic oil. The invention provides a method of treating a tissue comprising the following steps: (a) providing a composition comprising an enzyme of the invention, (b) providing a tissue; and (c) treating the tissue with the enzyme. Tissue treatment can include improved handling and fall of the final fabric, dye, obtaining flame retardancy, obtaining water repellency, obtaining optical brightener or obtaining resin finish. The fabric may comprise cotton, viscose, rayon, lyocell, flax fiber, linen, ramie, all blends thereof, or blends thereof with polyesters, wool, polyamide acrylics or polyacrylics. The invention provides a fabric, yarn or fiber comprising an enzyme of the invention. The enzyme can be adsorbed, absorbed or immobilized on the surface of the fabric, yarn or fiber. The invention provides methods for expressing phospholipase C comprising providing a Pichia strain with a Mut phenotype + ; inserting a nucleic acid encoding heterologous phospholipase C; and, culturing the Pichia strain under the conditions whereby phospholipase C is expressed. The method may also comprise supplementing the culture conditions with zinc. The invention also provides cell systems, isolated cells and cell strains for expressing phospholipase C comprising a Pichia strain of Mut phenotype + comprising a nucleic acid encoding heterologous phospholipase C operably linked to a promoter operable in the Pichia strain. The invention provides zeocin resistant yeast cell systems (e.g., yeast cells, cell strains, individual cells) for expressing a heterologous protein comprising the steps of providing a Pichia sp. (e.g. P. pastoris) comprising a heterologous nucleic acid capable of expressing a heterologous protein; culturing the cell under conditions comprising zeocin at an initial concentration; selecting cells resistant to the initial concentration of zeocin, and culturing under conditions comprising a higher concentration of zeocin; and selecting the cells grown in step (c) resistant to the highest concentration of zeocin. In one aspect, the heterologous protein is an enzyme, or optionally, a phospholipase, or optionally a phospholipase C (PLC), for example, any enzyme of the invention. Details of one or more embodiments of the invention are mentioned in the accompanying drawings and description below. Other features, objects and advantages of the invention will become evident from the description and drawings, and from the claims. All publications, patents, patent applications, GenBank sequences, and ATCC filings, cited herein are incorporated herein by reference for all purposes. Brief Description of Drawings The following drawings are illustrative of embodiments of the invention and are not meant to limit the scope of the invention when encompassed by the claims. Figure 1 is a block diagram of a computer system, as described in detail, below. Figure 2 is a flow diagram illustrating one aspect of a process 200 for comparing a new nucleotide or protein sequence to a sequence database in order to determine levels of homology between the new sequence and the sequences in the database , as described in detail below. Figure 3 is a flow diagram illustrating one embodiment of a process on a computer for determining whether two sequences are homologous, as described in detail below. Figure 4 is a flow diagram illustrating one aspect of an identifier process for detecting the presence of a feature in a sequence, as described in detail below. Figures 5A, 5B and 5C schematically illustrate a model two-phase system for PLC-mediated degumming simulation, as described in detail in Example 2, below. Figure 6 schematically illustrates an exemplary vegetable oil refining process employing the phospholipases of the invention. Figure 7 schematically illustrates an exemplary desizing process of the invention for physically refined oils, as discussed in detail, below. Figure 8 schematically illustrates phosphatide hydrolysis with a phospholipase C of the invention, as discussed in detail, below. figure 9 schematically illustrates a refinement process exemplary caustic agent of the invention, and illustrates an alternative embodiment comprising applying a phospholipase C of the invention as a "Caustic Refinement Aid" (Long Mixed Caustic Refinement), as discussed in detail, below. Figure 10 schematically illustrates the application of a phospholipase C of the invention as a desizing aid, as discussed in detail below. Figure 11 is a diagram depicting selected features of exemplary nucleic acids and polypeptides of the invention, as described in greater detail, below. Figure 12 schematically illustrates data from a two enzyme system of the invention, as described in Example 3, below. Figure 13 schematically illustrates an exemplary caustic refinement process of the invention, and illustrates an alternative embodiment comprising applying a phospholipase C of the invention as a "Caustic Refining Aid" (Long Mix Caustic Refining), as discussed in detail, below. Figure 14 illustrates another variation of methods of the invention where two centrifugation steps are employed in the process, as discussed in detail below. Figure 15 illustrates another variation of methods of the invention where three centrifugation steps are employed in the process, as discussed in detail below. Figure 16 illustrates another exemplary variation of this process employing acid treatment and having a centrifugation step prior to a desizing step, as discussed in detail, below. Figure 17 illustrates the results of in vitro digestion experiments on the phospholipase C variants of the invention, as discussed in detail in Example 4, below. Figure 18 illustrates the results of a batch fermenter culture employing an exemplary enzyme of the invention, as discussed in detail in Example 5, below. Figure 19 illustrates the results of Oxygen Uptake Rate ("TCO") comparisons of cultures of MutS strains of P. pastoris of the invention, as discussed in detail in Example 5, below. Figure 20 illustrates a comparison of methanol consumption profile in P. pastoris MutS strains of the invention, as discussed in detail in Example 5, below. Figure 21 illustrates a "TCO" profile of a culture of a recombinant form of the exemplary PLC enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 5, below. Figure 22 illustrates results from an SDS-PAGE showing the quality of PLC protein produced in a culture, and a corresponding TCO profile, of a culture of a recombinant form of the exemplary PLC enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 5, below. Figure 23 illustrates SDS-PAGE results showing the amount of active PLC located intracellularly in a culture of a recombinant form of the exemplary PLC enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 5, below. Figure 24 illustrates a visualization of the morphological changes in yeast cells associated with active PLC - a recombinant form of the exemplary PLC enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 5, below. Figure 25 graphically summarizes data showing the status of a 95 hr TFT PLC production performance (total fermentation time) in Pichia employing an exemplary PLC enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 5 , below. Figure 26 is a summary of the expression assessment data table of zeocin-adapted cell colonies of the invention, as discussed in detail in Example 5, below. Figure 27 illustrates data showing that PLC protein levels were higher in cultures comprising exemplary zeocin-adapted cell colonies of the invention, as discussed in detail in Ex. peep 5, below. Figure 28 illustrates data showing a comparison of growth of zeo-adapted colonies of the invention vs control, as discussed in detail in Example 5, below. Figure 29 illustrates the results of a heating experiment demonstrating the thermostability of the exemplary enzyme of SEQ ID NO: 2 of the invention, with the conditions indicated in the figure, as discussed in detail in Example 6, below. Figure 30 illustrates NMR data summarizing the heating experiment demonstrating the thermostability of the exemplary enzyme of SEQ ID NO: 2 of the invention, as discussed in detail in Example 6, below. Figures 31, 32 and 33 illustrate data demonstrating the thermal stability of SEQ ID NO: 2 employing p-NPPC, under the conditions shown in the figure, as discussed in detail in Example 6, below. Figure 34 illustrates data demonstrating the thermal stability of SEQ ID NO: 2 employing DSC analysis, as discussed in detail in Example 6, below. Similar reference symbols in the various drawings indicate similar elements. Detailed Description of the Invention The present invention provides phospholipases, for example, polypeptides that have phospholipase A, B, C, D, patatin, phosphatidic acid phosphatases (PAP) and / or lipid acyl hydrolase (LAH) or equivalent activity, polynucleotides coding them and methods for preparing and using them. The invention provides enzymes that effectively cleave the glycerolphosphate ester bond in oils, such as vegetable oils, for example oilseed phospholipids, to generate a water extractable phosphorylated base and a diglyceride. In one aspect, the phospholipases of the invention have a lipid acyl hydrolase (LAH) activity. In alternative aspects, the phospholipases of the invention can cleave glycerolphosphate ester bonds in phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid and / or sphingomyelin or a combination thereof. For example ie, in one aspect a phospholipase of the invention is specific for one or more specific substrates, for example, an enzyme of the invention may have a specificity of action for PE and PC; PE and PI; PE and PS; PS and PE; PS and PI; PI and PE; PS, PI and PC; PE, PI and PC; or PE, PS, PI and PC. A phospholipase of the invention (e.g. polypeptides having phospholipase A, B, C, D, patatin, phosphatidic acid phosphatases (PAP) and / or lipid acyl hydrolase (LAH) or equivalent activity) can be employed for enzymatic degumming of oils vegetables because the phosphate part is water soluble and easy to remove. The diglyceride product will remain in the oil and thus reduce losses. The PLCs of the invention can be employed in addition to or in place of PLA1s and PLA2s in commercial oil degumming, such as in the ENZYMAX® process where phospholipids are hydrolyzed by PLA1 and PLA2. In one aspect, the phospholipases of the invention are active at a high and / or a low temperature, or, over a wide temperature range, for example, they can be active at temperatures ranging from 20°C to 90°C, between 30°C to 80°C, or between 40°C to 70°C. The invention likewise provides phospholipases of the invention which have activity at alkaline pHs or at acidic pHs, for example low acidity in water. In alternative aspects, the phospholipases of the invention may have activity at acidic pHs as low as pH 6.5, pH 6.0, pH 5.5, pH 5.0, pH 4.5, pH 4.0 and pH 3, 5 or more acidic (i.e. < pH 3.5). In alternative aspects, the phospholipases of the invention may have activity at alkaline pHs as high as pH 7.5, pH 8.0, pH 8.5, pH 9.0, pH 9.5, pH 10 or more alkaline (i.e. , > pH 10). The phospholipases of the invention are active in the temperature range of about 40°C in one aspect, to about 70°C, 75°C, or 80°C, or more, under conditions of low activity in water (low water content). Water). The invention likewise provides methods for modifying exemplary phospholipases of the invention to generate enzymes with desirable properties. For example, phospholipases generated by the methods of the invention may have altered substrate specificities, substrate binding specificities, substrate cleavage patterns, thermal stability, pH / activity profile, pH / stability profile (such as increased stability at low eg pH values, PH<6 or pH<5, or high eg pH>9), stability to oxidation, dependence from ca 2+ , specific activity and the like. The invention provides alteration of any property of interest. For example, the alteration may result in a variant that, when compared to a phospholipase source, has an altered pH and temperature activity profile. In one aspect, the phospholipases of the invention are employed in various vegetable oil process steps, such as in vegetable oil extraction, particularly, in the removal of "phospholipid gums" in a process called "oil degumming", as described herein. . The invention provides compositions (e.g. comprising enzymes of the invention) and processes for producing vegetable oils from various sources, such as rice bran, soybeans, rapeseed, peanut, sesame, sunflower and corn oil. The phospholipase enzymes of the invention can be employed in place of PLA, for example phospholipase A2, in any vegetable oil processing step. Definitions The term "phospholipase" encompasses enzymes that have some phospholipase activity, for example, cleaving a glycerolphosphate ester bond (catalyzing hydrolysis of a glycerolphosphate ester bond), for example, in an oil, such as a vegetable oil . The phospholipase activity of the invention can generate a water-extractable phosphorylated base and a diglyceride. The phospholipase activity of the invention likewise includes hydrolysis of glycerol phosphate ester bonds at high temperatures, low temperatures, alkaline pHs and at acidic pHs. The term "a phospholipase activity" likewise includes the cleavage of a glycerol phosphate ester to generate a water-extractable phosphorylated base and a diglyceride. The term "a phospholipase activity" likewise includes the cleavage of glycerin and phosphoric acid ester bonds in phospholipids. The term "a phospholipase activity" likewise includes other activities, such as the ability to bind and hydrolyze a substrate, such as an oil, for example, a vegetable oil, substrate likewise including animal and plant phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines and sphingomyelins. The phospholipase activity may comprise a phospholipase C (PLC) activity; a phospholipase A (PLA) activity, such as a phospholipase A1 or phospholipase A2 activity; a phospholipase B (PLB) activity, such as a phospholipase B1 or phospholipase B2 activity, including lysophospholipase (LPL) activity and / or lysophospholipase transacylase (LPTA) activity; a phospholipase D (PLD) activity, such as a phospholipase D1 or a phospholipase D2 activity; and / or a patatin activity or any combination thereof. Phospholipase activity may comprise hydrolysis of a glycoprotein, for example as a glycoprotein found in a potato tuber or any plant of the Solanum genus, for example Tuberosum solarium. Phospholipase activity may comprise an enzymatic patatin activity, such as a patatin esterase activity (see, for example, Jimenez (2002) Biotechnol. Prog. 18:635-640 ). Phospholipase activity may comprise a lipid acyl hydrolase (LAH) activity. Phospholipase activity may comprise being specific for one or more specific substrates, for example an enzyme of the invention may have a specificity of action for PE and PC; PE and PI; PE and PS; PS and PE; PS and PI; PI and PE; PS, PI and PC; PE, PI and PC; or, PE, PS, PI and PC, or any combination thereof. In one aspect, a phospholipase of the invention may have multifunctional activity, for example, a combination of one or more of the enzyme activities described herein. For example, in one aspect, a polypeptide of the invention is enzymatically active, but requires lipase activity or requires any enzymatic activity that affects a neutral oil (triglyceride) fraction. It may be desirable to employ such a polypeptide in a particular process, for example in a desizing process where it is important that the neutral oil fraction is not impaired (decreased, degraded, e.g. hydrolyzed). Thus, in one aspect, the invention provides a desizing process comprising use of a polypeptide of the invention which has a phosphorylation activity. lipase, but not a lipase activity. In one aspect, PLC phospholipases of the invention utilize (e.g., catalyze hydrolysis of) a variety of phospholipid substrates including phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI) and / or phosphatidic acid or a combination of these. Furthermore, these enzymes may have varying degrees of activity on the lysophospholipid forms of these phospholipids. In various aspects, PLC enzymes of the invention may show a preference for phosphatidylcholine and phosphatidylethanolamine as substrates. In one aspect, phosphatidylinositol PLC phospholipases of the invention utilize a variety of phospholipid substrates including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and phosphatidic acid or a combination thereof. Furthermore, these enzymes may have varying degrees of activity on the lysophospholipid forms of these phospholipids. In various aspects, phosphatidylinositol PLC enzymes of the invention may show a preference for phosphatidylinositol as a substrate. In one aspect, the patatin enzymes of the invention utilize a variety of phospholipid substrates including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and phosphatidic acid, or a combination thereof. Furthermore, these enzymes may have varying degrees of activity on the lysophospholipid forms of these phospholipids. In several aspects, the patatins of the invention are based on a conservation of amino acid sequence similarity. In many respects, these enzymes exhibit a diverse set of biochemical properties and can carry out reactions characteristic of the PLA1, PLA2, PLC or PLD enzyme classes. In one aspect, the PLD phospholipases of the invention utilize a variety of phospholipid substrates including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and phosphatidic acid or a combination thereof. Furthermore, these enzymes may have varying degrees of activity on the lysophospholipid forms of these phospholipids. In one aspect, these enzymes are useful for carrying out transesterification reactions to produce structured phospholipids. The term "antibody" includes a peptide or polypeptide derived from, patterned after or substantially encoded by an immunoglobulin gene or immunoglobulin genes or fragments thereof, capable of specifically binding an antigen or epitope, see, for example, Fundamental Immunology, Third Edition, W.E. Paul, ed., Raven Press, N.Y. (1993); Wilson (1994) J. Immunol. Methods 175:267-273; Yarmush (1992) J. Biochem. Biophys. Methods 25:85-97. The term antibody includes antigen binding portions, i.e., "antigen binding sites", (e.g., fragments, subsequences, complementarity determining regions (CDRs)) that maintain the ability to bind antigen, including (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) an F(ab') 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single subdivision of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of a domain of VH; and (vi) an isolated complementarity determining region (CDR). Single chain antibodies are likewise included by reference in the term "antibody". The terms "array" or "microarray" or "biochip" or "chip" when used herein is a plurality of target elements, each target element comprising a defined amount of one or more polypeptides (including antibodies) or nucleic acids immobilized on a defined area of ​​a substrate surface, as discussed in more detail below. When used herein, the terms "computer," "computer program" and "processor" are used in their broadest general contexts and incorporate all such devices, as described in detail below. A "coding sequence" or a "coding sequence" for a particular polypeptide or protein is a nucleic acid sequence that is transcribed and translated into a polypeptide or protein when placed under the control of appropriate regulatory sequences. The term "expression cassette" as used herein refers to a nucleotide sequence that is capable of affecting the expression of a structural gene (i.e., a protein coding sequence, such as a phospholipase of the invention) in a compatible host. with such sequences. Expression cassettes include at least one promoter operably linked with the polypeptide coding sequence; and, optionally, with other sequences, for example transcription termination signals. Additional factors necessary or useful in carrying out the expression may likewise be employed, for example, enhancers. "Operably linked" as used herein refers to the binding of a promoter upstream from a DNA sequence such that the promoter mediates transcription of the DNA sequence. Thus, expression cassettes likewise include plasmids, expression vectors, recombinant viruses, any form of recombinant "naked DNA" vector, and the like. A "vector" comprises a nucleic acid that can infect, transfect, transiently or permanently transduce a cell. It will be recognized that a vector may be a naked nucleic acid or a nucleic acid complexed with a protein or lipid. The vector optionally comprises viral or bacterial proteins and / or nucleic acids, and / or membranes (e.g., a cell membrane, a viral lipid envelope, etc.). Vectors include, but are not limited to, replicons (eg, RNA replicons, bacteriophages) to which DNA fragments can be linked and become replicated. Vectors in this way include, but are not limited to, RNA, autonomous self-replication, linear or circular DNA or RNA (e.g., plasmids, viruses, and the like, see, e.g., U.S. Patent No. 5,217,879), and also includes expression and non-expression plasmids. Where a recombinant microorganism or cell culture is described as being host to an "expression vector" this also includes extrachromosomal linear and circular DNA and DNA that has been incorporated into the host chromosome(s). Where a vector is being maintained by a host cell, the vector can be stably replicated by the cells during mitosis as an autonomous structure, or be incorporated into the host genome. "Plasmids" are designated by a lowercase letter "p" preceded and / or followed by uppercase letters and / or numbers. The starting plasmids herein are commercially available, publicly available on an unrestricted basis, or may be constructed from available plasmids in accordance with published procedures. In addition, plasmids equivalent to those described herein are known in the art and will be apparent to the ordinarily skilled artisan. The term "gene" means the segment of DNA involved in the production of a polypeptide chain, including, among other things, regions preceding and following the coding region, such as leader and trailer, promoters and enhancers, as well as, where applicable, intermediate sequences (introns) between individual coding segments (exons). The phrases "nucleic acid" or "nucleic acid sequence" as used herein refer to an oligonucleotide, nucleotide, polynucleotide, or a fragment of any of these, for DNA or RNA (e.g., mRNA, rRNA, tRNA, iRNA ) of genomic or synthetic origin that can be single-stranded or double-stranded and can represent a sense or antisense strand, for peptide nucleic acid (PNA), or for any DNA-like or RNA-like material, natural or synthetic in origin, including e.g. iRNA, ribonucleoproteins (e.g. double-stranded iRNAs, e.g. iRNPs). The term encompasses nucleic acids, i.e. oligonucleotides, containing known analogues of natural nucleotides. The term likewise encompasses nucleic acid-like structures with synthetic backbones, see eg Mata (1997) Toxicol. App. Pharmacol. 144:189-197; Strauss-Soukup (1997) Biochemistry 36:8692-8698; Samstag (1996) Antisense Nucleic Acid Drug Dev 6:153-156. "Amino acid" or "amino acid sequence" as used herein refers to an oligopeptide, peptide, polypeptide or protein sequence, or in a fragment, portion or subunit of any of these, and in synthetic or naturally occurring molecules. The terms "polypeptide" and "protein" when used herein refer to amino acids linked together by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain modified amino acids other than the 20 amino acids encoded per gene. . The term "polypeptide" likewise includes peptides and polypeptide fragments, motifs and the like. The term likewise includes glycosylated polypeptides. The peptides and polypeptides of the invention likewise include all "mimetic" and "peptidomimetic" forms, as described in more detail, below. When used herein, the term "isolated" means that the material is removed from its original environment (eg, natural environment if naturally occurring). For example, a naturally occurring polypeptide or polynucleotide present in a living animal is not isolated, but the same polynucleotide or polypeptide, separated from some or all of the materials coexisting in the natural system, is isolated. Such polynucleotides may form part of a vector and / or such polynucleotides or polypeptides may form part of a composition, and yet being isolated in such a vector or composition is not part of its natural environment. When used herein, an isolated composition or material may likewise be a "purified" composition, that is, it does not require absolute purity; rather, it is intended as a relative definition. Individual nucleic acids obtained from a library can be purified conventionally to electrophoretic homogeneity. In alternative aspects, the invention provides nucleic acids that have been purified from genomic DNA or other sequences in a library or other environment by at least one, two, three, four, five or more orders of magnitude. When used herein, the term "recombinant" means that the nucleic acid is adjacent to a "main chain" nucleic acid to which it is not adjacent in its natural environment. In one aspect, nucleic acids represent 5% or more of the number of insertions of nucleic acid in a population of nucleic acid "backbone molecules". "Backbone molecules" according to the invention include nucleic acids such as such as expression vectors, self-replicating nucleic acids, viruses, integrating nucleic acids and similar vectors or nucleic acids employed to maintain or manipulate a nucleic acid insert of interest. In one aspect, the enriched nucleic acids represent 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more of the number of nucleic acid insertions in the population of chain molecules. main recombination. "Recombinant" proteins or polypeptides refer to those polypeptides or proteins produced by recombinant DNA techniques; for example, produced from cells transformed by an exogenous DNA construct encoding the desired protein or polypeptide. "Synthetic" protein or polypeptides are those prepared by chemical synthesis, as described in more detail below. A promoter sequence is "operably linked to" a coding sequence when RNA polymerase that initiates transcription at the promoter will transcribe the coding sequence into mRNA, as discussed further below. "Oligonucleotide" refers to a single-stranded polydeoxynucleotide or two complementary polydeoxynucleotide strands that can be chemically synthesized. Such synthetic oligonucleotides have no 5' phosphate and therefore will not bind to another oligonucleotide without adding a phosphate with an ATP in the presence of a kinase. A synthetic oligonucleotide will bind to a fragment that has not been dephosphorylated. The phrase "substantially identical" in the context of two nucleic acids or polypeptides, refers to two or more sequences that have at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% , 98% or 99% amino acid or nucleotide residue (sequence) identity, when compared and aligned for maximum match, when measured employing any known sequence comparison algorithm, as discussed in detail below, or by visual inspection. In alternative aspects, the invention provides polypeptide and nucleic acid sequences having significant identity to an exemplary sequence of the invention, for example, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc., over a region of at least about 100 residues, 150 residues, 200 residues, 300 residues, 400 residues, or a region ranging from about 50 residues to the natural size of the nucleic acid or polypeptide. Nucleic acid sequences of the invention can be substantially identical over the natural length of a polypeptide coding region. Additionally, a "substantially identical" amino acid sequence is a sequence that differs from a reference sequence by one or more conservative or non-conservative amino acid substitutions, deletions, or insertions, particularly when such a substitution occurs at a site that is not the active site of the molecule, and as long as the polypeptide essentially maintains its functional properties. A conservative amino acid substitution, for example, substitutes one amino acid for another of the same class (e.g., substitution of one hydrophobic amino acid, such as isoleucine, valine, leucine, or methionine, for another, or substitution of one polar amino acid for another, such as as a substitution of arginine for lysine, glutamic acid for aspartic acid, or glutamine for asparagine). One or more amino acids can be deleted, for example, from a phospholipase polypeptide, resulting in modification of the polypeptide's structure, without significantly altering its biological activity. For example, carboxyl- or amino-terminal amino acids that are not required for biological activity can be removed. Modified polypeptide sequences of the invention can be analyzed for phospholipase biological activity by any number of methods, including contacting the modified polypeptide sequence with a phospholipase substrate and determining whether the modified polypeptide decreases the amount of specific substrate in the assay or increases the bioproducts. of the enzymatic reaction of a functional phospholipase with the substrate, as discussed further below. "Hybridization" refers to the process by which a nucleic acid strand joins with a complementary strand by base pairing. Hybridization reactions can be sensitive and selective in a that a particular sequence of interest can be identified even in samples in which it is present in low concentrations. Appropriately stringent conditions can be defined, for example, by the concentrations of salt or formamide in the prehybridization and hybridization solutions, or by the hybridization temperature, and are well known in the art. For example, stringency can be increased by lowering the salt concentration, increasing the formamide concentration, or raising the hybridization temperature, altering the hybridization time, as described in detail below. In alternative aspects, nucleic acids of the invention are defined by their ability to hybridize under various stringency conditions (eg, high, medium, and low), as mentioned herein. The term "variant" refers to polynucleotides or polypeptides of the invention modified at one or more base pairs, codons, introns, exons or amino acid residues (respectively) yet still retain the biological activity of a phospholipase of the invention. Variants can be produced by any number of means including methods such as, for example, error prone PCR, scrambling, oligonucleotide-directed mutagenesis, assembly PCR, sexual PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, ensemble mutagenesis recursive, exponential ensemble mutagenesis, site-specific mutagenesis, gene reassembly, GSSM® and any combination thereof. Techniques for producing variant phospholipases having activity at a pH or temperature, for example, that is different from a wild-type phospholipase, are included herein. The term "saturation mutagenesis", Gene Site Saturation Mutagenesis® (GSSM®) or "GSSM®" includes a method that uses degrading oligonucleotide primers to introduce point mutations into a polynucleotide, as described in detail below. The term "optimized directed evolution system" or "optimized directed evolution" includes a method for reassembling fragments of related nucleic acid sequences, eg related genes, and clarified in detail, below. The term "synthetic ligation reassortment" or "SLR" includes a method of ligating oligonucleotide fragments in a non-stochastic manner, and is explained in detail below. Generation and Manipulation of Nucleic Acids The invention provides isolated and recombinant nucleic acids (e.g., SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO : 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155 , SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ Exemplary ID NO: 173), including expression cassettes such as expression vectors, encoding the polypeptides and phospholipases of the invention. The invention likewise includes methods for discovering novel phospholipase sequences employing the nucleic acids of the invention. Also provided are methods for modifying the nucleic acids of the invention by, for example, synthetic linkage reassembly, optimized directed evolution system, and / or saturation mutagenesis. The nucleic acids of the invention can be made, isolated and / or manipulated by, for example, cloning and expression of cD-NA libraries, PCR message or genomic DNA amplification, and the like. By practicing the methods of the invention, homologous genes can be modified by manipulating a standard nucleic acid, as described herein. The invention may be practiced in conjunction with any method or protocol or device known in the art, which is well described in the scientific and patent literature. General Techniques The nucleic acids employed to practice this invention, whether RNA, iRNA, antisense nucleic acid, cDNA, genomic DNA, vectors, viruses or hybrids thereof, can be isolated from a variety of sources, genetically engineered, amplified and / or expressed / recombinantly generated. Recombinant polypeptides generated from these nucleic acids can be individually isolated or cloned and tested for a desired activity. Any recombinant expression system may be employed, including bacterial, mammalian, yeast, insect, or plant cell expression systems. Alternatively, these nucleic acids can be synthesized in vitro by known chemical synthesis techniques, as described in, for example, Adams (1983) J. Am. Chem. social 105:661; Belousov (1997) Nucleic Acids Res. 25:3440-3444; Frenkel (1995) Free Radic.Biol. Med. 19:373-380; Blommers (1994) Biochemistry 33:7886-7896; Narang (1979) Meth. En-zymol. 68:90; Brown (1979) Meth. Enzymol. 68:109; Beaucage (1981) Tetra. Lett. 22:1859; U.S. Patent At the. 4,458,066. Techniques for handling nucleic acids, such as, for example, subcloning, labeling probes (for example, random primer labeling employing Klenow polymerase, notch translation, amplification), sequencing, hybridization, and the like are well described in the scientific literature. and patent, see, for example, Sambrook, ed., Molecular Cloning: A Laboratory Manual (2 o ed.), vols. 1-3, Cold Spring Harbor Laboratory, (1989); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, Aiisubel, ed. John Wiley & Sons, Inc., New York (1997); LABORATORY TECHNIQUES IN BIOCHEMISTRY AND MOLECULAR BIOLOGY: HYBRIDIZATION WITH NUCLEIC ACID PROBES, Part I. Theory and Nucleic Acid Preparation, Tijssen, ed. Elsevier, N.Y. (1993). Other useful means of obtaining and manipulating nucleic acids employed to practice the methods of the invention is to clone from genomic samples, and, if desired, to evaluate and re-clone isolated or amplified inserts from, for example, genomic clones or cDNA clones. Nucleic acid sources employed in the methods of the invention include cDNA or genomic libraries contained in, for example, mammalian artificial chromosomes (MACs), see, for example, U.S. Patent. Us. 5,721,118; 6,025,155; human artificial chromosomes, see, for example, Rosenfeld (1997) Nat. Gene 15:333-335; yeast artificial chromosomes (YAC); bacterial artificial chromosomes (BAC); P1 artificial chromosomes, see, for example, Woon (1998) Genomics 50:306-316; P1-derived vectors (PACs), see, for example, Kern (1997) Biotechniques 23:120-124 ; cosmids, recombinant viruses, phages or plasmids. In one aspect, a nucleic acid encoding a polypeptide of the invention is assembled in appropriate phase with a leader sequence capable of directing the secretion of the translated polypeptide or fragment thereof. The invention provides fusion proteins and nucleic acids encoding them. A polypeptide of the invention can be fused to a heterologous polypeptide or peptide, such as N-terminal identification peptides that give desired characteristics, such as increased stability or simplified purification. Peptides and polypeptides of the invention may likewise be synthesized and expressed as fusion proteins with one or more additional domains linked thereto to, for example, produce a more immunogenic peptide, to more easily isolate a recombinantly synthesized peptide, to identify and isolating antibodies and antibody-expressing B cells, and the like. Purification and detection facilitating domains include, for example, metal-chelation peptides such as polyistidine tracts and histidine-tryptophan modules that allow for purification. immobilized metals, protein A domains that allow purification into immobilized immunoglobulin, and the domain used in the FLAGS affinity / extension purification system (Immunex Corp, Seattle WA). The inclusion of a cleavable linker sequence such as Factor Xa or enterokinase (Invitrogen, San Diego CA) between a purification domain and the motif-comprising polypeptide or peptide to facilitate purification. For example, an expression vector may include a nucleic acid sequence encoding epitope linked to six histidine residues followed by a thioredoxin and an enterokinase cleavage site (see, for example, Williams (1995) Biochemistry 34:1787- 1797; Dobeli (1998) Protein Expr. Pu-rif. 12:404-414). The histidine residues facilitate detection and purification while the enterokinase cleavage site provides a means of purifying the epitope from the remainder of the fusion protein. Technology pertaining to vectors encoding fusion proteins and application of fusion proteins is well described in the scientific and patent literature, see for example Kroll (1993) DNA Cell. Biol., 12:441-53. Translational and transcriptional control sequences The invention provides nucleic acid (e.g. DNA) sequences of the invention operably linked to expression control sequence(s) (e.g. transcriptional or translational), e.g. promoters or enhancers, to direct or modulate synthesis / RNA expression. The expression control string can be in an expression vector. Exemplary bacterial promoters include lacI, lacZ, T3, T7, gpt, lambda PR, PL and trp. Exemplary eukaryotic promoters include early i-mediate CMV, HSV thymidine kinase, early and late SV40, retrovirus LTRs, and mouse metallothionein I. Promoters suitable for expressing a polypeptide in bacteria include the E. coli lac or trp promoters, the lacI promoter, the lacZ promoter, the T3 promoter, the T7 promoter, the gpt promoter, the lambda PR promoter, the lambda PL promoter, promoters of operons encoding glycolytic enzymes such as 3-phosphoglycerate kinase (PGK), and the acid phosphatase promoter. Eukaryotic promoters include the promoter CMV immediate early, HSV thymidine kinase promoter, heat shock promoters, the SV40 early and late promoter, retrovirus LTRs, and the mouse metallothionein-1 promoter. Other promoters known to control gene expression in prokaryotic or eukaryotic cells or their viruses may likewise be employed. Expression vectors and cloning vehicles The invention provides expression vectors and cloning vehicles comprising nucleic acids of the invention, for example, sequences encoding the phospholipases of the invention. Expression vectors and cloning vehicles of the invention may comprise viral particles, baculovirus, phage, plasmids, phagemids, cosmids, phosmids, bacterial artificial chromosomes, viral DNA (e.g. vaccinia, adenovirus, contagious epithelioma virus, pseudorabies and derived from SV40), P1-based artificial chromosomes, yeast plasmids, yeast artificial chromosomes and any other specific vectors for specific hosts of interest (such as Bacillus, Aspergillus and yeast). Vectors of the invention may include chromosomal, non-chromosomal and synthetic DNA sequences. Large numbers of suitable vectors are known to those skilled in the art and are commercially available. Exemplary vectors include: bacterial: pQE vectors (Qiagen), pBluescript plasmids, pNH vectors, (lambda-ZAP vectors (Stratagene); p-trc99a, pKK223-3, pDR540, pRIT2T (Pharmacia); Eukaryotic: pXT1, pSG5 (Stratagene), pSVK3, pBPV, pMSG, pSVLSV40 (Pharmacia) However, any other plasmid or other vector can be used as long as they are replicable and viable in the host. Low copy number or high copy number vectors can be used employed with the present invention. The expression vector may comprise a promoter, a ribosome binding site for translation initiation and a transcriptional terminator. The vector may also include appropriate sequences to amplify expression. Mammalian expression vectors may comprise an origin of replication, any required ribosome binding sites saries, a polyadenylation site, splice donor and acceptor sites, transcriptional termination sequences and 5' flanking untranscribed sequences. In some aspects, DNA sequences derived from the SV40 junction and polyadenylation sites can be employed to provide the required non-transcribed genetic elements. In one aspect, the expression vectors contain one or more selectable marker genes to allow selection of host cells containing the vector. Such selectable markers include genes encoding dihydrofolate reductase or genes that confer neomycin resistance for eukaryotic cell culture, genes that confer resistance to tetracycline or ampicillin in E. coli, and the S. cerevisiae TRP1 gene. Promoter regions can be selected from any desired gene employing chloramphenicol transferase (CAT) vectors or other vectors with selectable markers. Vectors for expressing the polypeptide or fragment thereof in eukaryotic cells may contain enhancers to increase expression levels. Enhancers are cis-acting elements of DNA, typically about 10 to about 300 bp in length that act on a promoter to increase its transcription. Examples include the SV40 enhancer on the 100 to 270 base pair late side of the origin of replication, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the lagging side of the origin of replication, and the adenovirus enhancers. A DNA sequence can be inserted into a vector by a variety of procedures. In general, the DNA sequence is ligated at the desired position in the vector following digestion of the insert and vector with appropriate restriction endonucleases. Alternatively, blunt ends on both the insert and the vector can be ligated. A variety of cloning techniques are known in the art, for example as described in Ausubel and Sambrook. Such procedures and the like are considered to be within the scope of those skilled in the art. The vector may be in the form of a plasmid, a viral particle or a phage. Other vectors include chromosomal DNA sequences co, nonchromosomal and synthetic, derived from SV40; bacterial plasmids, phage DNA, baculovirus, yeast plasmids, vectors derived from combinations of plasmids and phage DNA, viral DNA such as vaccinia, adenovirus, avian pox and pseudorabies. A variety of expression and cloning vectors for use with prokaryotic and eukaryotic hosts are described by, for example, Sambrook. Particular bacterial vectors that may be employed include commercially available plasmids comprising genetic elements from the well-known cloning vector pBR322 (ATCC 37017), pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden), GEM1 (Promega Biotec, Madison, WI, USA). ) pQE70, pQE60, pQE-9 (Qiagen), pD10, psiX174 pBluescript II KS, pNH8A, pNH16a, pNH18A, pNH46A (Stratagene), ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 (Pharmacia), pKK232-8 and pCM7. Particular eukaryotic vectors include pSV2CAT, pOG44, pXT1, pSG (Stratagene) pSVK3, pBPV, pMSG and pSVL (Pharmacia). However, any other vector can be used as long as it is replicable and viable in the host cell. Host cells and transformed cells The invention likewise provides a transformed cell comprising a nucleic acid sequence of the invention, for example a sequence encoding a phospholipase of the invention, a vector of the invention. The host cell can be any of the host cells familiar to those skilled in the art, including prokaryotic cells, eukaryotic cells, such as bacterial cells, fungal cells, yeast cells, mammalian cells, insect cells, or plant cells. Enzymes of the invention may be expressed in any host cell, for example any bacterial cell, any yeast cell, for example Pichia Pastoris, Saccharomyces cerevisiae or Schizosaccharomyces pombe. Exemplary bacterial cells include E. coli, Lactococcus lac-tis, Streptomyces, Bacillus subtilis, Bacillus cereus, Salmonella typhimurium or any species within the genera Bacillus, Streptomyces and Staphylococcus. Exemplary insect cells include Drosophila S2 and Spodopte- ra Sf9. Exemplary animal cells include CHO, COS or Bowes melanoma or any mouse or human cell strain. Selection of an appropriate host is within the capabilities of those skilled in the art. The vector can be introduced into host cells using any of a variety of techniques, including transformation, transfection, transduction, viral infection, gene guns, or Ti-mediated gene transfer. Particular methods include calcium phosphate transfection, DEAE-Dextran mediated transfection, lipofection or electroporation (Davis, L., Dibner, M., Battey, I., Basic Methods in Molecular Biology, (1986)). Where appropriate, constructed host cells can be cultured in conventional nutrient media modified where appropriate to activate promoters, select transformants, or amplify the genes of the invention. Following transformation of an appropriate host strain and growth of the host strain to an appropriate cell density, the selected promoter can be induced by appropriate means (e.g., temperature change or chemical induction) and the cells can be cultured for a period of time. additional period to allow them to produce the desired polypeptide or fragment thereof. Cells can be harvested by centrifugation, disrupted by physical or chemical means, and the resulting crude extract kept for further purification. Microbial cells employed for protein expression can be disrupted by any convenient method, including freeze-thaw cycling, sonication, mechanical disruption, or use of cell lysing agents. Such methods are well known to those skilled in the art. The expressed polypeptide or fragment thereof can be recovered and purified from recombinant cell cultures by methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin. Protein reduplication steps can be employed, when necessary, in completing the polypeptide configuration. If desired, high performance liquid chromatography (HPLC) can be employed for final purification steps. Various mammalian cell culture systems can also be employed to express recombinant protein. Examples of mammalian expression systems include monkey kidney fibroblast COS-7 strains and other cell strains capable of expressing compatible vector proteins, such as C127, 3T3, CHO, HeLa and BHK cell strains. The construct in host cells can be employed in a conventional manner to produce the gene product encoded by the recombinant sequence. Depending on the host employed in a recombinant production procedure, polypeptides produced by host cells containing the vector may be glycosylated or may be non-glycosylated. Polypeptides of the invention may or may not likewise include an initial methionine amino acid residue. Cell-free translational systems can likewise be employed to produce a polypeptide of the invention. Cell-free translational systems can use mRNAs transcribed from a DNA construct comprising a promoter operably linked to a nucleic acid encoding the polypeptide or fragment thereof. In some aspects, the DNA construct can be linearized prior to conducting an in vitro transcription reaction. The transcribed mRNA is then incubated with an appropriate cell-free translation extract, such as a rabbit reticulocyte extract, to produce the desired polypeptide or fragment thereof. Expression vectors may contain one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells such as dihydrofolate reductase or neomycin resistance for eukaryotic cell culture or such as tetracycline or ampicillin resistance in E. coli. An exemplary phospholipase C enzyme (having a sequence as mentioned in SEQ ID NO: 2) was over-expressed in active form in a variety of host systems including gram negative bacteria such as E. coli, gram positive bacteria such as any Bacillus sp. (e.g. Bacillus subtilis, Bacillus cereus), yeast host cells (including, for example, Pichia pastoris, Saccharomyces sp., such as S. cerevisiae and S. pombe) and Lactococcus lactis, or mammalian, fungal, plant cells or insect. The active enzyme is expressed from a variety of constructs in each host system. These nucleic acid expression constructs may comprise nucleotides encoding the full-length open reading frame (composed of the signal sequence, the prosequence and the mature protein coding sequence) or they may comprise a subset of these genetic elements. alone or in combination with heterologous genetic elements that serve as the signal sequence and / or the prosequence of the mature open reading frame. Each of these systems can serve as a commercial production host for the expression of PLC for use in the previously described enzymatic oil degumming processes. Amplification of Nucleic Acids In the practice of the invention, nucleic acids encoding the polypeptides of the invention, or modified nucleic acids, can be reproduced by, for example, amplification. The invention provides amplification primer sequence pairs for amplifying nucleic acids encoding polypeptides with a phospholipase activity. In one aspect, primer pairs are capable of extending nucleic acid sequences of the invention, for example, including exemplary SEQ ID NO: 1, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 3, or a subsequence thereof; a sequence as mentioned in SEQ ID NO: 5, or a subsequence thereof; and, a sequence as mentioned in SEQ ID NO: 7, or a subsequence thereof, etc. One skilled in the art can design amplification primer sequence pairs for any part of or the full length of these sequences. The invention provides an amplification primer sequence pair for amplifying a nucleic acid encoding a polypeptide having a phospholipase activity, wherein the primer pair is capable of amplifying a nucleic acid comprising a sequence of the invention, or fragments or subsequences thereof. One or each member of the amplification primer sequence pair may comprise an oligonucleotide comprising at least about 10 to 50 consecutive bases of sequence, or about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 consecutive bases of sequence. The invention provides amplification primer pairs, wherein the primer pair comprises a first member having a sequence as mentioned by about the first (the 5') 12, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 23, 24, or 25 residues of a nucleic acid of the invention and a second member having a sequence as mentioned by about the first (the 5') 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 residues of the complementary strand of the first member. The invention provides phospholipases generated by amplification, for example, polymerase chain reaction (PCR), employing an amplification primer pair of the invention. The invention provides methods of preparing a phospholipase by amplification, for example, polymerase chain reaction (PCR), employing an amplification primer pair of the invention. In one aspect, the amplification primer pair amplifies a nucleic acid from a library, for example, a gene library, such as an environmental library. Amplification reactions can also be employed to quantify the amount of nucleic acid in a sample (such as the amount of message in a cell sample), label the nucleic acid (e.g., apply it to an array or stain), detect nucleic acid or quantify the amount of a specific nucleic acid in a sample. In one aspect of the invention, message isolated from a cell or a cDNA library is amplified. The experienced technician can select and design suitable oligonucleotide amplification primers squares. Amplification methods are likewise well known in the art, and include, for example, polymerase chain reaction, PCR (see, for example, PCR PROTOCOLS, A GUIDE TO METHODS AND APPLICATIONS ed. Innis, Academic Press, N.Y. (1990) ) and PCR STRATEGIES 5 (1995), ed. Innis, Academic Press, Inc., N.Y., ligase chain reaction (LCR) (see, for example, Wu (1989) Genomics 4:560; Landegren (1988) Science 241:1077; Barringer (1990) Gene 89:117); transcriptional amplification (see, for example, Kwoh (1989) Proc. Natl. Acad. Sci. USA 86:1173); and, self-sustaining sequence replication (see, for example, Guatelli (1990) Proc. Natl. 10 Acad. Know. USA 87:1874); Q Beta replicase amplification (see, e.g., Smith (1997) J. Clin. Microbiol. 35:1477-1491), automatic Q-beta replicase amplification assay (see, e.g., Burg (1996) Mol. Cell Probes 10:257-271) and other RNA polymerase-mediated techniques (e.g., NASBA, Cangene, Mississauga, Ontario); see also Berger 15 (1987) Methods Enzymol. 152:307-316; Sambrook; Ausubel; U.S. Patent Us. 4,683,195 and 4,683,202; Sooknanan (1995) Biotechnology 13:563-564. Determining the degree of sequence identity The invention provides isolated and recombinant nucleic acids comprising sequences having at least about 50%, 51%, 52%, 20 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or full sequence identity (100%) for an exemplary nucleic acid of the invention (for example, SEQ ID NO: 1, SEQ ID NO: 3, SEO ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO : 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID 30 NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, and nucleic acids encoding SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO : 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56 , SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO : 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106 , SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO: 174) over a region of at least about 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550 or more residues. The invention provides polypeptides comprising sequences having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater, or complete sequence identity (100%) for a exemplary polypeptide of the invention. The extent of sequence identity (homology) can be determined using any computer program and associated parameters, including those described herein, such as BLAST 2.2.2. or FASTA version 3.0t78, with the basic parameters. In alternative embodiments, the sequence identification may be over a region of at least about 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400 consecutive residues, or natural length. of the nucleic acid or polypeptide. The extent of sequence identity (homology) can be determined using any computer program and associated parameters, including those described herein, such as BLAST 2.2.2. or FASTA version 3.0t78, with the basic parameters. Figure 11 is a diagram depicting selected features of exemplary nucleic acids and polypeptides of the invention, including sequence identity comparison of exemplary sequences from public databases. All sequences described in Figure 11 were subjected to a BLAST search (as described in detail, below) against two sets of databases. The first set of database is available through NCBI (National Center for Biotechnology Information). All the results of searches against these databases are found in the columns entitled "Description of NR", "Accession Code NR", "Value E of NR" or "Organismo de NR". "NR" refers to the Non-Redundant Nucleotide Database maintained by NCBI. This database is a combination of GenBank, GenBank updates, and tions. Entries in the "NR Description" column refer to the definition line in any given NCBI record, which includes a description of the sequence, such as the source organism, gene name / protein name, or some description of the sequence's function. . The entries in the "NR Accession Code" column refer to the unique identifier determined in a sequence record. The entries in the column "E value of NR" refer to the Expected value (E value), which represents the probability that an alignment score as good as that found between the sequence in question (the sequences of the invention) and a sequence from the bank of data would be found in the same number of comparisons between random sequences as was done in the present BLAST survey. The entries in the "NR Organism" column refer to the source organism of the sequence identified as the closest BLAST hit. The second group of databases is collectively known as the Geneseq® database, which is available from Thomson Derwent (Philadelphia, PA). All results of searches against this database are found in the columns titled "Description of Geneseq Protein", "Accession Code of Geneseq Protein", "E-Value of Geneseq Protein", "Description of Geneseq DNA", "Geneseq DNA Accession Code" or "Geneseq DNA E-value". The information found in these columns is comparable to the information found in the NR columns described above, unless it was derived from BLAST searches against the GeneseqTM database rather than the NCBI databases. In addition, this table includes the "No. of EC Predicted" column. An EC number is the number assigned to a type of enzyme according to a standardized enzyme nomenclature scheme developed by the Enzyme Commission of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB). The results in the "No. of Predicted EC" column are determined by a BLAST search against the Kegg database (Kyoto Encyclopedia of Genes and Genomes). If the top BLAST comparison has an E value equal to or less than e' 6 , the EC number assigned to the top pair is entered in the table. The EC number of top stroke is used as a guide to where the EC number of the sequence of the invention might be. The columns "Length of DNA in Question" and "Length of Protein in Question" refer to the number of nucleotides or the number of amino acids, respectively, in the sequence of the invention that was searched or interrogated against the Geneseq databases or NCBI The "NR or Geneseq DNA Length" and "NR or Geneseq Protein Length" columns refer to the number of nucleotides or the number of amino acids, respectively, in the top pair sequence of the BLAST search search. The results provided in these columns are from the survey that declared the lowest E value, from the NCBI databases, or from the Geneseq database. The columns "Geneseq or NR %ID Protein" and "Geneseq or NR %ID DNA" refer to the percent of sequence identity between the sequence of the invention and the sequence of the top BLAST pair. The results provided in these columns are from the survey that declared the lowest E value, either from the NCBI databases or the Geneseq database. Homologous sequences likewise include RNA sequences in which uridines replace thymines in nucleic acid sequences. Homologous sequences can be obtained using any of the procedures described here or can result from correcting a sequencing error. It will be appreciated that nucleic acid sequences as mentioned here may be represented in the only traditional character format (see, for example, Stryer, Lubert. Biochemistry, 3 a Ed., W.H Freeman & Co., New York) or in any other format that records the identity of nucleotides in a sequence. Various sequence comparison programs identified herein are employed in this aspect of the invention. Nucleic acid and / or protein sequence identities (homologies) can be assessed employing any of a variety of sequence comparison programs and algorithms known in the art. Such algorithms and programs include, but are not limited to, TBLASTN, BLASTP, FASTA, TFASTA, and CLUSTALW (Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85(8):2444 - 2448, 1988; Altschul and the like, J. Mol. Biol. 215(3):403-410, 1990; Thompson et al., Nucleic Acids Res. 22(2):4673-4680, 1994; Higgins and the like, Methods Enzymol. 266:383-402, 1996; Altschul and the like, J. Mol. Biol. 215(3):403-410, 1990; Altschul and the like, Nature Genetics 3: 266-272, 1993). Homology or identity can be measured using sequence analysis software (eg, Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wl 53705). Such software compares to similar sequences by designating degrees of homology in various deletions, substitutions and other modifications. The terms "homology" and "identity" in the context of two or more nucleic acid or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides. which are the same when compared and aligned for maximum correspondence over a comparison window or designated region when measured employing any number of sequence comparison algorithms or by manual alignment and visual inspection. For sequence comparison, a sequence can act as a reference sequence (an exemplary sequence SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc.) in which test sequences are compared. When employing a sequence comparison algorithm, reference and test sequences are recorded in a computer, subsequence coordinates are assigned, if necessary, and sequence algorithm program parameters are assigned. Default program parameters can be used, or alternative parameters can be assigned. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters. A "comparison window", as used herein, includes reference to a segment of any number of contiguous residues. For example, in alternative aspects of the invention, contiguous residues ranging in part from 20 to the natural length of an exemplary sequence of the invention, for example, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO : 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc., are compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Whether the reference sequence has the required sequence identity for an exemplary sequence of the invention, e.g. 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% , 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more identity of sequence for a sequence of the invention, for example SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc., that sequence is within the scope of the invention. In alternative embodiments, subsequences ranging from about 20 to 600, about 50 to 200, and about 100 to 150 are compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Sequence alignment methods for comparison are well known in the art. Optimal alignment of sequences for comparison can be conducted, for example, by the local homology algorithm of Smith & Waterman, Adv. App. Math. 2:482, 1981, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443, 1970, for research on the similarity method of person & Lipman, Proc. Nat'l. academy Sci. USA 85:2444, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA at Wisconsin Genetics Software Pa-chage, Genetics Computer Group, 575 Science Dr., Madison, WI), or by manual alignment and visual inspection . Other algorithms for determining homology or identity include, for example, in addition to a BLAST (Basic Local Alignment Search Tool at the National Center for Biological Informatrion) program, ALIGN, AMAS (Analysis of Multiply Aligned Se quences), AMPS (Protein Multiple Sequence Alignment), ASSET (Aligned Segment Statistical Evaluation Tool), BANDS, BESTSCOR, BIOSCAN (Biological Sequence Comparative Analysis Node), BLIMPS (BLocks IMProve Searcher), FASTA, Intervals & Points, BMB, CLUSTAL V , CLUSTAL W, CONSENSUS, LCONSENSUS, WCONSENSUS, Smith-Waterman algorithm, DARWIN, Las Vegas algorithm, FNAT (Forced Nucleotide Alignment Tool), Framealign, Framesearch, DINAMIC, FILTER, FSAP (Fris-tensky Sequence Analysis Package), GAP (Global Alignment Program), GENAL, GIBBS, GenQuest, ISSC (Sensitive Sequence Comparison), LA-LIGN (Local Sequence Alignment), LCP (Local Content Program), MACAW (Multiple Alignment Construction & Analysis Workbench), MAP (Multiple Alignment Program ), MBLKP, MBLKN, PIMA (Pattern-Induced MultiSequence Alignment), SAGA (Sequence Alignment by Genetic Algorithm) and WHAT-IF. Such alignment programs can likewise be employed to evaluate genome databases to identify polynucleotide sequences having substantially identical sequences. Several genome databases are available, for example, a substantial portion of the human genome is available as part of the Human Genome Sequencing Project (Gibbs, 1995). Several genomes have been sequenced, for example, M. genitalium (Fraser et al., 1995), M. jannaschii (Bult et al., 1996), H. influenzae (Fleischmann et al., 1995), E. coli (Blattner et al., 1995), E. coli (Blattner et al., 1996). , 1997), and yeast (S. cerevisiae) (Mewes et al., 1997), and D. melanogaster (Adams et al., 2000). Significant progress was also made in sequencing the genomes of model organisms, such as mice, C. elegans, and Arabadopsis sp. Databases containing genomic information annotated with some functional information are maintained by a different organization, and are accessible via the internet. BLAST, BLAST 2.0 and BLAST 2,2,2 algorithms are likewise employed to practice the invention. They are described, for example, in Altschul (1977) Nuc. Acids Res. 25:3389-3402; Altschul (1990) J. Mol. Biol. 215:403-410. The software to perform BLAST analysis is available publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high-score string pairs (HSPs) by identifying short words of length W in the string in question, which compares or satisfies some threshold score of positive value T when aligned with a word of the same length in a string of strings. data base. T is referred to as the neighbor word score threshold (Altschul (1990) supra). These initial neighborhood word hits act as families to initiate searches to find longer HSPs containing them. These word strokes are extended in both directions along each sequence until the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the M parameters (reward score for a pair of comparison residues; always >0). For amino acid sequences, a score matrix is ​​used to calculate the cumulative score. The extension of word strokes in each direction is stopped when: the cumulative alignment score drops by the amount X from its maximum obtained value; the cumulative score starts from zero or below, due to the accumulation of one or more negative score residue alignments; or at the end of each sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLAST program (for nucleotide sequences) defaults to a word length (W) of 11, an expectation (E) of 10, M=5, N= -4, and a comparison of both strands. For amino acid sequences, the BLASTP program defaults to a word length of 3, and expectations (E) of 10, and the BLOSUM62 score matrix (see Henikoff & Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915) alignments (B) of 50, expectation (E) of 10, M=5, N= -4, and a comparison of both filaments. The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin & Altschul (1993) Proc. Natl. A-cad. Sci. USA 90:5873). An algorithm-provided measure of similarity of BLAST is the smallest sum probability (P(N)) that gives an indication of the probability by which a pair between two nucleotide or amino acid sequences would occur by change. For example, a nucleic acid is considered similar to a reference sequence if the probability of 5 sums less in a comparison of test nucleic acid to nucleic acid. reference is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001. In one aspect, nucleic acid and protein sequence homologies are evaluated employing the Basic Local Alignment Search Tool ("BLAST"). For example, five specific BLAST programs can be employed to perform the following tasks: (1) BLASTP and BLAST3 compare a given amino acid sequence against a protein sequence database; (2) BLASTN compares a given nucleotide sequence against a nucleotide sequence database; (3) BLASTX 15 compares the conceptual translational products of six structures of a nucleotide sequence in question (both strands) against a protein sequence database; (4) TBLASTN compares a protein sequence in question against a translated nucleotide sequence database in all six reading frames (both 20 strands); and, (5) TBLASTX compares the translations of six structures of a nucleotide sequence in question together with the translations of six structures from a nucleotide sequence database. BLAST programs identify homologous sequences by identifying similar segments, which are referred to herein as "high-scoring segment pairs," between a nucleic acid or amino sequence in question and a test sequence that is preferably obtained from a nucleic acid or protein sequence database. High scoring segment pairs are preferably identified (i.e., aligned) by means of a score matrix, many of which are known in the art. Preferably, the score matrix employed is the BLOSUM62 matrix (Gonnet and the like, Science 256:1443-1445, 1992; Henikoff and Henikoff, Proteins 17:49-61, 1993). Less preferably, the PAM or PAM250 arrays can in the same way to be employed (see, for example, Schwartz and Dayhoff, eds., 1978, Matrices for Detections Distance Relationships: Atlas of Protein Sequence and Structure, Washington: National Biomedical Research Foundation). In one aspect of the invention, to determine whether a nucleic acid has the required sequence identity to be within the scope of the invention, the NCBI BLAST 2.2.2 programs are employed. Default options for blastp. There are about 38 placement options in the BLAST 2.2.2 program. In this exemplary aspect of the invention, all default values ​​are employed except for setting default filtering (ie, all parameters are set to default except filtering which is set to OFF); instead a "-F F" setting is employed, which disables filtration. Use of default filtering often results in Karlin-Altschul violations due to the short length of the sequence. The default values ​​employed in this exemplary aspect of the invention, and to determine the values ​​in Figure 11, as discussed above, include: "Filter for low complexity: ON > Word Size: 3 > Matrix: Blosum62 > Space Costs: Existence: 11 > Extension: 1" Other default settings are: filter for low complexity OFF, word size of 3 for protein, matrix of BLOSUM62, space existence penalty of -11 and a space extension penalty of -1. An exemplary NCBI BLAST 2.2.2 program placement is mentioned in Example 1, below. Note that the "-W" option is missing 0. This means that, if not fixed, the word length is missing by 3 for proteins and 11 for nucleotides. Computer systems and computer program products To determine and identify sequence identities, structural homologies, motifs and the like in silico, a nucleic acid sequence The co or polypeptide of the invention can be stored, recorded and manipulated in any medium that can be read and accessed by a computer. In this way, the invention provides computers, computer systems, computer readable media, computer program products and the like recorded or stored in such polypeptide and nucleic acid sequences of the invention, e.g., an exemplary sequence of the invention, e.g., SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc. When used herein, the words "recorded" and "stored" refer to a process for storing information on a computer medium. A skilled artisan can readily adopt any known methods for recording information on a computer readable medium to generate fabrications comprising one or more of the polypeptide and / or nucleic acid sequences of the invention. Another aspect of the invention is a computer readable medium having at least one nucleic acid and / or polypeptide sequence of the invention recorded thereon. Computer readable media include magnetically readable media, optically readable media, electronically readable media, magnetic / optical media, flash memory. For example, computer readable media may be a hard disk, floppy disk, magnetic tape, flash memory, CD-ROM, Digital Versatile Disk (DVD), Random Access Memory (RAM), Read Only Memory (ROM), ), or any type of means known to those skilled in the art. Aspects of the invention include systems (eg, internet-based systems), particularly computer systems, that store and manipulate the sequences and sequence information described herein. An example of a computer system 100 is illustrated in block diagram form in Figure 1. When used herein, "a computer system" refers to the hardware components, software components, and data storage components employed to analyze a polypeptide or nucleotide sequence of the invention. The system computer 100 may include a processor for processing, accessing and manipulating the sequence data. Processor 105 can be any well-known type of central processing unit, such as, for example, the Pentium III from Intel Corporation, or similar processor from Sun, Motorola, Compaq, AMD or International Business Machines. Computer system 100 is a general purpose system comprising processor 105 and one or more internal data storage components 110 for storing data, and one or more data retrieval devices for retrieving data stored in data storage components. Dice. A skilled technician can easily appreciate that any of the currently available computer systems are suitable. In one aspect, computer system 100 includes a processor 105 connected to a bus that is connected to main memory 115 (preferably implemented as RAM) and one or more internal data storage devices 110, such as a hard drive and / or or other computer readable media having data recorded thereon. Computer system 100 may also include one or more data retrieval devices 118 for reading data stored on internal data storage devices 110. Data retrieval device 118 may represent, for example, a floppy disk drive, a compact disk drive, a magnetic tape drive, or a modem capable of connecting to a remote data storage system (e.g., over the Internet). ) etc. In some embodiments, the internal data storage device 110 is a removable computer readable medium such as a floppy disk, a compact disc, a magnetic tape, etc. containing control logic and / or data recorded therein. The computer system 100 may advantageously include or be programmed by appropriate software to read control logic and / or data from the data storage component once inserted into the data retrieval device. Computer system 100 includes a display screen 120 which is employed to display output to a computer user. He must- it is also noted that computer system 100 may be linked to other computer systems 125a-c in a wide area network or network to provide centralized access to computer system 100. Software for accessing and processing the amino acid or nucleotide sequences of the invention may reside in main memory 115 during execution. In some aspects, computer system 100 may also comprise a sequence comparison algorithm for comparing a nucleic acid sequence of the invention. The algorithm and sequence(s) may be stored on a computer readable medium. A "sequence comparison algorithm" refers to one or more programs that are implemented (locally or remotely) in the computer system 100 to compare a nucleotide sequence with other nucleotide sequences and / or compounds stored within a medium. of data storage. For example, the sequence comparison algorithm can compare the nucleotide sequences of an exemplary sequence, for example, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO : 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, etc. stored on computer readable medium reference sequences stored on computer readable medium to identify homologies or structural motifs. The parameters used with the algorithms above can be adapted depending on the length of the sequence and the degree of homology studied. In some respects, the parameters may be the default parameters employed by algorithms in the absence of user instructions. Figure 2 is a flow diagram illustrating one aspect of a process 200 for comparing a new protein or nucleotide sequence to a sequence database in order to determine levels of homology between the new sequence and the sequences in the database. . The sequence database may be a private database stored within the computer system 100, or a public database such as GENBANK which is available over the Internet. Process 200 starts at a source state 201 and then moves to a state 202 in that the new string to be compared is stored in a memory on a computer system 100. As discussed above, the memory could be any type of memory, including RAM or an internal storage device. Process 200 then moves to a state 204 where a database of sequences is opened for analysis and comparison. Process 200 then moves to a state 206 where the first sequence stored in the database is read into memory in the computer. A comparison is then performed at a state 210 to determine whether the first sequence is the same as the second sequence. It is important to note that this step is not limited to performing an exact comparison between the new sequence and the first sequence in the database. Well known methods are known to those skilled in the art to compare two protein or nucleotide sequences, even if they are not identical. For example, spaces can be introduced into a sequence in order to increase the level of homology between the two tested sequences. The parameters that control whether spaces or other features are introduced in a sequence during the comparison are normally entered by the user of the computer system. Once a comparison of the two sequences has been performed at state 210, a determination is made at decision state 210 whether the two sequences are the same. Of course, the term "same" is not limited to sequences that are absolutely identical. Sequences that are within the homology parameters entered by the user will be marked as "same" in process 200. If a determination is made that the two sequences are the same, process 200 moves to a state 214 where the sequence name of the database is displayed to the user. This state notifies the user that the string with the displayed name meets the homology constraints that have been entered. Once the name of the stored sequence is displayed to the user, the process 200 moves to a decision state 218 where a determination is made whether more sequences exist in the database. If no further sequence exists in the database, then process 200 ends up in a final state 220. However, if more sequences exist in the database, then process 200 moves to a state 224 where a pointer is moved to the next sequence in the database so that it can be compared to the new sequence. In this way, the new sequence is aligned and compared with every sequence in the database. It should be noted that if a determination were made in decision state 212 that the sequences were not homologous, then process 200 would immediately move to decision state 218 in order to determine whether any other sequences are available in the database for Comparation. Thus, one aspect of the invention is a computer system comprising a processor, a data storage device having stored therein a nucleic acid sequence of the invention, and a sequence comparator for conducting the comparison. The sequence comparator can indicate a level of homology between the compared sequences or identify structural motifs, or it can identify structural motifs in sequences that are compared to those nucleic acid codes and polypeptide codes. Figure 3 is a flow diagram illustrating one embodiment of a process 250 on a computer for determining whether two sequences are homologous. Process 250 starts at a source state 252 and then moves to a state 254 where a first sequence to be compared is stored in a memory. The second string to be compared is then stored in memory at state 256. Process 250 then moves to a state 260 where the first character in the first string is read and then to a state 262 where the first character of the second string is read. It should be understood that if the sequence is a nucleotide sequence, then the character would normally be A, T, C, G, or U. If the sequence is a protein sequence, then it may be a single-letter amino acid code of so that the first and second sequences can be easily compared. A determination is then made in a decision state 264 if the two characters are the same. If they are the same, then process 250 moves to a state 268 where the next characters in the first and second strings are read. A determination is then made if the next characters are the same. If they are, then process 250 continues this loop until the two characters are not the same. If a determination is made that the next two characters are not the same, process 250 moves to a decision state 274 to determine if there are any more characters in each string to read. If there are no more characters to read, then the process 250 moves to a state 276 where the level of homology between the first and second strings is displayed to the user. The level of homology is determined by calculating the proportion of characters between sequences that were the same out of the total number of sequences in the first sequence. Thus, if every character in a first nucleotide sequence 100 aligned with all characters in a second sequence, the homology level would be 100%. Alternatively, the computer program may compare a reference sequence to a sequence of the invention to determine whether the sequences differ in one or more positions. The program can record the length and identity of substituted, deleted or inserted amino acid residues or nucleotides with respect to the sequence of the reference or the invention. The computer program may be a program that determines whether a reference sequence contains a single nucleotide polymorphism (SNP) with respect to a sequence of the invention, or, whether a sequence of the invention comprises an SNP of a known sequence. Thus, in some respects, the computer program is a program that identifies SNPs. The method can be implemented by the computer systems described above and the method illustrated in Figure 3. The method can be carried out by reading a sequence of the invention and the reference sequences through the use of the computer program and identifying differences with the program of computer. In other respects the computer-based system with comprises an identifier for identifying features within a nucleic acid or polypeptide of the invention. An "identifier" refers to one or more programs that identify certain characteristics within a nucleic acid sequence. For example, an identifier may comprise a program that identifies an open reading frame (ORF) in a nucleic acid sequence. Figure 4 is a flow diagram illustrating one aspect of an identifier process 300 for detecting the presence of a feature in a sequence. Process 300 starts at a source state 302 and then moves to a state 304 in which a first sequence that will be checked for characteristics is stored to a memory 115 in computer system 100. Process 300 then moves to a state 306 where a database of sequence characteristics is opened. Such a database would include a list of each feature attribute along with the feature name. For example, a feature name could be "Codon Initiation" and the attribute would be "ATG." Another example would be the feature name "Caixa TAATAA". and the feature attribute would be 'TAATAA'. An example of such a database is produced by the University of Wisconsin Genetics Computer Group. Alternatively, the features may be structural polypeptide motifs such as alpha helices, beta laminae, or polypeptide motifs functional such as enzymatic active sites, helix-turn-helix motifs or other motifs known to those skilled in the art As soon as the characteristics database is opened at state 306, process 300 moves to a state 308 where the first characteristic is read from the database. A comparison of the attribute of the first characteristic with the first sequence is then made in a state 310. A determination is then made in a decision state 316 whether the attribute of the characteristic was found in the first sequence. If the attribute was found, then the process 300 moves to a state 318 where the found feature name is displayed given to the user. The process 300 then moves to a decision state 320 in which a determination is made whether feature motion exists in the data base. If no more features exist, then process 300 terminates in a final state 324. However, if more features exist in the database, then process 300 reads the next sequence feature at a state 326 and resumes at state 310 where the attribute of the next characteristic is compared against the first sequence. If the feature attribute is not found in the first sequence in decision state 316, process 300 moves directly to decision state 320 in order to determine if any more features exist in the database. Thus, in one aspect, the invention provides a computer program that identifies open reading frames (ORFs). A polypeptide or nucleic acid sequence of the invention can be stored and manipulated in a variety of data processing programs in a variety of formats. For example, a string can be stored as text in a word processing file, such as MicrosoftWORD or WORDPERFECT, or as an ASCII file in a variety of database programs familiar to those skilled in the art, such as DB2, SYBASE, or ORACLE Furthermore, many computer and database programs can be employed as sequence comparison algorithms, identifiers or sources of reference nucleotide sequences or polypeptide sequences to be compared to a nucleic acid sequence of the invention. The programs and databases employed to practice the invention include, but are not limited to: MacPattern (EMBL), DiscoveryBase (Molecular Applications Group), GeneMine (Molecular Applications Group), Look (Molecular Applications Group), MacLook (Molecular Applications Group ), BLAST and BLAST2 (NCBI), BLASTN and BLASTX (Altschul and the like, J., Mol. Biol. 215: 403, 1990), FASTA (Pearson and Lipman, Proc. Natl. Acad. Sci. USA, 85: 2444 , 1988), FASTDB (Brutlag and the like Comp. App. Biosci. 6:237-245, 1990), Catalyst (Molecular Simulations Inc.), Catalyst / SHAPE (Molecular Simulations Inc.), Cerius2.DBAccess (Molecular Simulations Inc. ), HypoGen (Molecular Simulations Inc.), Insight II, (Molecular Simulations Inc.), Discover (Mo Lecular Simulations Inc.), CHARMm (Molecular Simulations Inc.), Felix (Molecular Simulations Inc.), DelPhi, (Molecular Simulations Inc.), QuanteMM, (Molecular Simulations Inc.), Homology (Molecular Simulations Inc.), Modeler ( Molecular Simulations Inc.), ISIS (Molecular Simulations Inc.), Quant-ta / Protein Design (Molecular Simulations Inc.), WebLab (Molecular Simulations Inc.), WebLab Diversity Explorer (Molecular Simulations Inc.), Gene Explorer (Molecular Simulations Inc.), SeqFold (Molecular Simulations Inc.), the MDL Available Chemicals Directory database, the MDL Drug Data Report data base, the Comprehensive Medicinal Chemistry database, the Derwent's World database Drug Index, the BioByteMasterFile database, the Genbank database and the Genseqn database. Many other programs and databases would be apparent to one skilled in the art given the present disclosure. Motifs that can be detected using the above programs include sequences encoding leucine zippers, helix-turn-helix motifs, glycosylation sites, ubiquitination sites, alpha helices, and beta laminae, signal sequences encoding signal peptides that direct secretion of encoded proteins, sequences involved in transcriptional regulation such as homeoboxes, acidic stretches, enzymatic active sites, substrate binding sites and enzymatic cleavage sites. Nucleic acid hybridization The invention provides recombinant or isolated nucleic acids that hybridize under stringent conditions to an exemplary sequence of the invention, for example, the sequence as mentioned in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, or a nucleic acid encoding a polypeptide comprising the sequence as mentioned in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO: 174. Severe conditions can be highly severe conditions , medium severe conditions, low severe conditions, including the high and low severity conditions described here. In alternative embodiments, nucleic acids of the invention as defined by their ability to hybridize under stringent conditions can be between about five residues and the natural size of the molecule, for example, an exemplary nucleic acid of the invention. For example, they can be at least 5, 10, 15, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80, 90, 100, 150, 200, 250, 300, 350, 400 or more residues in length. Nucleic acids shorter than full-length are likewise included. These nucleic acids are useful as, for example, hybridization probes, labeling probes, PCR oligonucleotide probes, iR-NA (single or double stranded), antisense or antibody binding peptide (epitope) encoding sequences. , motifs, active sites, binding domains, regulatory domains and the like. In one aspect, nucleic acids of the invention which are defined by their ability to hybridize under high stringency comprise conditions of about 50% formamide at about 37°C to 42°C. In one aspect, nucleic acids of the invention are defined by their ability to hybridize under reduced stringency comprising conditions in about 35% to 25% formamide at about 30°C to 35°C. Alternatively, nucleic acids of the invention are defined by their ability to hybridize under high stringency comprising conditions at 42°C in 50% formamide, 5X SSPE, 0.3% SDS, and a repetitive sequence blocking nucleic acid, such as DNA of salmon sperm or cot-1 (eg 200 n / ml denatured or sheared salmon sperm). In one aspect, nucleic acids of the invention are defined by their ability to hybridize under reduced stringency conditions comprising 35% formamide at a reduced temperature of 35°C. Following hybridization, the filter can be washed with 6X SSC, 0.5% SDS at 50°C. These conditions are considered to be conditions "moderate" conditions above 25% formamide and "low" conditions below 25% formamide. A specific example of "moderate" hybridization conditions is when the above hybridization is conducted in 30% formamide. A specific example of "low stringency" hybridization conditions is when the above hybridization is conducted in 10% formamide. The temperature range corresponding to a particular level of severity can also be reduced by calculating the purine to pyrimidine ratio of the nucleic acid of interest and adjusting the temperature in this way. Nucleic acids of the invention are likewise defined by their ability to hybridize under high, medium, and low stringency conditions as mentioned in Ausubel and Sambrook. Variations from the above ranges and conditions may be employed to practice the invention and are well known in the art. Hybridization conditions are discussed as well, below. Oligonucleotide probes and methods for employing them The invention likewise provides nucleic acid probes to identify nucleic acids encoding a polypeptide having a phospholipase activity. In one aspect, the probe comprises at least 10 consecutive bases of a sequence as mentioned in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEO ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO : 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151 , SEQ ID NO: 153, SEQ ID NO; 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO; 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173. Alternatively, a probe of the invention can be at least about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80, 90, 100, or 150, or more, or about 10 to 50, about 20 to 60 about 30 to 70, consecutive bases of a sequence as mentioned in a section - sequence of invention. The probes identify a nucleic acid by binding or hybridization. Probes may be employed in arrangements of the invention, see discussion below, including, for example, capillary arrangements. The probes of the invention may likewise be employed to isolate other nucleic acids or polypeptides. The probes of the invention can be employed to determine whether a biological sample, such as a soil sample, contains an organism having a nucleic acid sequence of the invention or an organism from which the nucleic acid was obtained. In such procedures, a biological sample potentially harboring the organism from which the nucleic acid was isolated is obtained and nucleic acids are obtained from the sample. Nucleic acids are contacted with the probe under conditions that allow the probe to specifically hybridize to any complementary sequence present in the sample. Where necessary, conditions that allow the probe to specifically hybridize to complementary sequences can be determined by contacting the probe with complementary sequences from samples known to contain the complementary sequence, as well as control sequences that do not contain the complementary sequence. Hybridization conditions, such as the salt concentration of the hybridization buffer, the formamide concentration of the hybridization buffer, or the hybridization temperature, can be varied to identify stand conditions that allow the probe to specifically hybridize to complementary nucleic acids (see discussion on specific hybridization conditions). If the sample contains the organism from which the nucleic acid was isolated, specific hybridization of the probe is then detected. Hybridization can be detected by labeling the probe with a detectable agent such as a radioactive isotope, a fluorescent dye, or an enzyme capable of catalyzing the formation of a detectable product. Many methods for using labeled probes to detect the presence of complementary nucleic acids in a sample are familiar to those skilled in the art. These include Southern blots, Northern blots, colony hybridization procedures, and spot stains. Protocols for each of these procedures are provided in Ausubel and Sambrook. Alternatively, more than one probe (at least one that is capable of specifically hybridizing to any complementary sequences that are present in the nucleic acid sample) can be employed in an amplification reaction to determine whether the sample contains an organism containing a sequence of nucleic acid of the invention (e.g., an organism from which the nucleic acid has been isolated). In one aspect, the probes comprise oligonucleotides. In one aspect, the amplification reaction may comprise a PCR reaction. PCR protocols are described in Ausubel and Sambrook (see discussion on amplification reactions). In such procedures, nucleic acids are contacted in the sample with the probes, the amplification reaction is performed, and any resulting amplification products are detected. The amplification product can be detected by performing gel electrophoresis on the reaction products and staining the gel with an intercalator such as ethidium bromide. Alternatively, one or more probes can be labeled with a radioactive isotope and the presence of a radioactive amplification product can be detected by autoradiography after gel electrophoresis. Probes derived from sequences near the 3' or 5' ends of a nucleic acid sequence of the invention can similarly be derived in chromosome walking procedures to identify clones containing additional, eg genomic, sequences. Such methods allow the isolation of genes encoding additional proteins of interest from the host organism. In one aspect, the nucleic acid sequences of the invention are employed as probes to identify and isolate related nucleic acids. In some aspects, similarly identified related nucleic acids may be cDNAs or genomic DNAs from organisms other than those from which the nucleic acid of the invention was first isolated. In such procedures, a nucleic acid sample is contacted with the probe under conditions that allow the probe to specifically hybridize to related sequences. Hybridization of the probe to nucleic acids of the related organism is then detected using any of the methods described above. In nucleic acid hybridization reactions, the conditions employed to obtain a particular level of stringency will vary, depending on the nature of the nucleic acids to be hybridized. For example, the length, degree of complementarity, nucleotide sequence composition (eg, GC v. AT content), and nucleic acid type (eg, RNA v. DNA) of the nucleic acid hybridization regions can be considered by selecting hybridization conditions. An additional consideration is whether one of the nucleic acids is immobilized, for example on a filter. Hybridization can be performed under low stringency, moderate stringency or high stringency conditions. As an example of nucleic acid hybridization, a polymer membrane containing immobilized denatured nucleic acids is first prehybridized for 30 minutes at 45°C in a solution consisting of 0.9 M NaCl, 50 mM NaH2PO4, pH 7, 0.5.0 mM Na 2 EDTA, 0.5% SDS, 10X Denhardt, and 0.5 mg / ml polyriboadenilic acid. Approximately 2 X 10 7 cpm (specific activity 4-9 X 10 8 cpm / ug) of labeled oligonucleotide probe per end of 32 P are then added to the solution. After 12-16 hours of incubation, the membrane is washed for 30 minutes at room temperature. (RT) in 1X SET (150 mM NaCl, 20 mM Tris hydrochloride, pH 7.8, 1 mM Na2EDTA) containing 0.5% SDS, followed by a 30 minute wash in 1X SET fresh at Tm-10°C for the oligonucleotide probe. The membrane is then exposed to the autoradiographic film for detection of hybridization signals. By varying the stringency of the hybridization conditions employed to identify nucleic acids, such as cDNAs or genomic DNAs, that hybridize to a detectable probe, nucleic acids having different levels of homology to the probe can be identified and isolated. The stringency can be varied by conducting hybridization at varying temperatures under the melting temperatures of the probes. The melting temperature, Tm, is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly complementary probe. Very severe conditions are selected to be equal to or about 5°C lower than the Tm for a particular probe. The probe melting temperature can be calculated using the following exemplary formulas. For probes between 14 and 70 nucleotides in length, the melting temperature (Tm) is calculated using the formula: Tm=81.5+16.6 (log [Na+])+0.41 (G+C fraction)-( 600 / N) where N is the probe length. If the hybridization is carried out in a solution containing formamide, the melting temperature can be calculated using the equation: Tm=81.5+16.6 (log [Na+])+0.41 (fraction G+C)-(0 .63% formamide)-(600 / N) where N is the length of the probe. Prehybridization can be performed in 6X SSC, 5X Denhardt's reagent, 0.5% SDS, 100pg denatured fragmented salmon sperm DNA or 6X SSC, 5X Denhardt's reagent, 0.5% SDS , 100pg denatured fragmented salmon sperm DNA, 50% formamide. Formulas for SSC and Denhardt and other solutions are listed, for example, in Sambrook. Hybridization is conducted by adding the detectable probe to the prehybridization solutions listed above. Where the probe comprises double-stranded DNA, it is denatured prior to addition to the hybridization solution. The filter is contacted with the hybridization solution for a sufficient period of time to allow the probe to hybridize to cDNAs or genomic DNAs containing sequences complementary thereto or homologous thereto. For probes over 200 nucleotides in length, hybridization can be performed at 15-25°C below the Tm. For shorter probes, such as oligonucleotide probes, hybridization can be conducted at 5-10°C below the Tm. In one aspect, 6X SSC hybridizations are conducted at approximately 68°C. In one aspect, hybridizations in 50% formamide containing solutions are conducted at approximately 42°C. All previous hybridizations would be considered to be under high stringency conditions. Following hybridization, the filter is washed to remove any detectable probe not specifically bound. The stringency employed to wash the filters can likewise be varied depending on the nature of the nucleic acids to be hybridized, the length of the nucleic acids to be hybridized, the degree of complementarity, the nucleotide sequence composition (e.g., content of GC v. AT), and the type of nucleic acid (eg RNA v. DNA). Examples of progressively higher stringency condition washes are as follows: 2X SSC, 0.1% SDS at room temperature for 15 minutes (low stringency -20°); 0.1X SSC, 0.5% SDS at room temperature for 30 minutes in 1 hour (moderate severity); 0.1X SSC, 0.5% SDS for 15 to 30 minutes between annealing temperature and 68°C (high stringency); and 0.15M NaCl for 15 minutes at 72°C (very high stringency). A final low stringency wash can be conducted in 0.1X SSC 25 at room temperature. The above examples are merely illustrative of a group of conditions that can be employed to practice the invention, for example, to wash filters. One skilled in the art would know that there are numerous recipes for washes of different severity, all of which can be employed to practice the invention. 30 Nucleic Acids That Hybridize to Probe Can Be Identified by autoradiography or other conventional techniques. The above procedure can be modified to identify nucleic acids having levels decreasing homology to the probe sequence. For example, to obtain nucleic acids of decreasing homology to the detectable probe, less stringent conditions may be employed. For example, the hybridization temperature can be lowered in 5°C increments from 68°C to 42°C in a hybridization buffer having a Na+ concentration of approximately 1M. Following hybridization, the filter can be washed with 2X SSC, 0.5% SDS at hybridization temperature. These conditions are considered to be "moderate" conditions above 50°C and "low" conditions below 50°C. An example of "moderate" hybridization conditions is when the above hybridization is conducted at 55°C. An example of "low stringency" hybridization conditions is when the above hybridization is conducted at 45°C. Alternatively, hybridization can be performed in buffers, such as 6X SSC, containing formamide at a temperature of 42°C. In this case, the concentration of formamide in the hybridization buffer can be reduced by 5% increments from 50% to 0% to identify clones having decreasing levels of homology to the probe. Following hybridization, the filter can be washed with 6X SSC, 0.5% SDS at 50°C. These conditions are considered to be "moderate" conditions above 25% formamide and "low" conditions below 25% formamide. A specific example of "moderate" hybridization conditions is when the above hybridization is conducted at 30% formamide. A specific example of "low stringency" hybridization conditions is when the above hybridization is conducted in 10% formamide. These probes and methods of the invention can be employed to isolate nucleic acids having a sequence of at least about 99%, by 95%, by 75%, by minus 98%, minus 90%, minus 70%, at least at least at least 97%, by 85%, by 65%, by minus 96%, minus 80%, minus 60%, at least at least at least 55%, or at least 50% homology to a nucleic acid sequence of the invention comprising at least about 10, 15, 20, 25, 30, 35, 40, 50, 75, 100, 150, 200, 250, 300, 350, 400, or 500 consecutive bases vas of this, and the sequences complementary thereto. Homology can be measured employing an alignment algorithm, as discussed here. For example, homologous polynucleotides may have a coding sequence that is a naturally occurring allelic variant of one of the coding sequences described herein. Such allelic variants may have a substitution, deletion or addition of one or more nucleotides when compared to the nucleic acids of the invention. Additionally, the probes and methods of the invention can be employed to isolate nucleic acids that encode polypeptides having at least about 99%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75% , at least 70%, at least 65%, at least 60%, at least 55%, or at least 50% sequence identity (homology) for a polypeptide of the invention comprising at least 5, 10, 15, 20, 25 , 30, 35, 40, 50, 75, 100, or 150 consecutive amino acids thereof as determined employing a sequence alignment algorithm (for example, such as the FASTA algorithm version 3.0t78 with default parameters, or a BLAST 2.2 program .2 with exemplary placements as mentioned here). Expression of Phospholipase Inhibition The invention also provides nucleic acids complementary to (eg antisense sequences) the nucleic acids of the invention, for example phospholipase-encoding nucleic acids. Antisense sequences are capable of inhibiting the transport, entanglement or transcription of genes encoding phospholipase. Inhibition can be performed by targeting genomic DNA or messenger RNA. Transcription or targeted nucleic acid function can be inhibited, for example, by hybridization and / or cleavage. A particularly useful group of inhibitors provided by the present invention include oligonucleotides which are capable of binding to phospholipase gene or message, in each case preventing or inhibiting phospholipase enzyme production or function. The association may be through sequence-specific hybridization. Another useful class of inhibitors includes oligonucleotides that cause inactivation or phosphorylation of messages. follipase. The oligonucleotide may have enzymatic activity that causes such cleavage, such as ribozymes. The oligonucleotide may be chemically modified or conjugated to an enzyme or composition capable of cleaving the complementary nucleic acid. One can rate a lake of many such different oligonucleotides for those with the desired activity. Inhibition of phospholipase expression may have a variety of industrial applications. For example, inhibition of phospholipase expression can reduce or prevent spoilage. Deterioration can occur when lipids or polypeptides, for example structural lipids or polypeptides, are enzymatically degraded. This can lead to spoilage, or rot, of fruits and vegetables. In one aspect, the use of compositions of the invention that inhibit phospholipase expression and / or activity, for example antibodies, antisense oligonucleotides, ribozymes and RNAi, are employed to reduce or prevent spoilage. Thus, in one aspect, the invention provides methods and compositions comprising applying to a plant or plant product (e.g., a fruit, seed, root, leaf, etc.) antibodies, antisense oligonucleotides, ribozymes, and RNAi of the invention. to reduce or prevent deterioration. These compositions may likewise be expressed by the plant (e.g., a transgenic plant) or another organism (e.g., a bacterium or other microorganism transformed with a phospholipase gene of the invention). Compositions of the invention for inhibiting phospholipase expression (e.g., antisense, iRNA, ribozymes, antibodies) can be employed as pharmaceutical compositions. Antisense Oligonucleotides The invention provides antisense oligonucleotides capable of binding a phospholipase message that can inhibit phospholipase activity by targeting mRNA. Strategies for designing antisense oligonucleotides are well described in the patent and scientific literature, and the skilled artisan can design such phospholipase oligonucleotides employing the novel reagents of the invention. For example, RNA mapping / gene walking protocols to assess for effective antisense oligonucleotides zes are well known in the art, see, for example, Ho (2000) Methods Enzymol. 314:168-183, describing an RNA mapping assay, which is based on standard molecular techniques to provide an easy and safe method for potent antisense sequence selection. See similarly Smith (2000) Eur. J. Pharm. Know. 11:191-198. Naturally occurring nucleic acids are used as antisense o-ligonucleotides. Antisense oligonucleotides can be of any length; for example, in alternative aspects, the antisense oligonucleotides are between about 5 to 100, about 10 to 80, about 15 to 60, about 18 to 40. The optimal length can be determined by routine screening. Antisense oligonucleotides can be present at any concentration. The optimal concentration can be determined by routine assessment. A wide variety of synthetic non-naturally occurring nucleotide and nucleic acid analogues are known that can address this potential problem. For example, peptide nucleic acids (PNAs) containing non-ionic backbones such as N-(2-aminoethyl) glycine units can be employed. Antisense oligonucleotides having phosphorothioate linkages may likewise be employed, as described in WO 97 / 03211; WO 96 / 39154; Mata (1997) Toxicol Appl Pharmacol 144:189 197; Antisense Therapeutics, ed. Agrawal (Humana Press, Totowa, N.J., 1996). Antisense oligonucleotides having synthetic DNA backbone analogues provided by the invention may likewise include phosphorodithioate, methylphosphonate, phosphoramidate, alkyl phosphotriester, sulfamate, 3'-thioacetal, methylene(methylimino), 3'-N -carbamate, and morpholine carbamate nucleic acids, as described above. Combinatorial chemistry methodology can be employed to create vast numbers of oligonucleotides that can be easily evaluated for specific oligonucleotides that have appropriate binding affinities and specificities for any target, such as the antisense and sense phospholipase sequences of the invention (see, for example, Gold (1995) J. of Biol. Chem. 270:13581-13584). Inhibitory Ribozymes The invention provides ribozymes capable of binding phospholipase message that can inhibit phospholipase enzymatic activity by targeting mRNA. Strategies for designing ribozymes and selecting the specific phospholipase antisense sequence for targeting are well described in the patent and scientific literature, and the skilled artisan can design such ribozymes employing the novel reagents of the invention. Ribozymes act by binding to a target RNA by the target RNA binding portion of a ribozyme that is held in close proximity to an enzymatic portion of the RNA that cleaves the target RNA. In this way, the ribozyme recognizes and binds a target RNA through complementary base pairing, and once bound to the correct site, acts enzymatically to cleave and inactivate the target RNA. Cleavage of a target RNA in such a manner will destroy its ability to direct synthesis of an encoded protein if the cleavage occurs in the coding sequence. After a ribozyme has bound and cleaved its target RNA, it is typically released from that RNA and thereby binds and cleaves new targets repeatedly. In some circumstances, the enzymatic nature of a ribozyme can be advantageous over other technologies, such as antisense technology (where a nucleic acid molecule simply binds to a nucleic acid target to block its transcription, translation, or association with another molecule). molecule) when the effective concentration of ribozyme required to carry out a therapeutic treatment may be lower than that of an antisense oligonucleotide. This potential advantage reflects the ability of the ribozyme to act enzymatically. Thus, a single ribozyme molecule is capable of cleaving many target RNA molecules. Furthermore, a ribozyme is typically a highly specific inhibitor, with the specificity of inhibition not only depending on the mechanism of base pairing binding, but also on the mechanism by which the molecule inhibits expression of the RNA to which it binds. . That is, inhibition is caused by cleavage of the target RNA and thus specificity is defined as the ratio of the rate of cleavage of targeted RNA to the rate of cleavage of untargeted RNA. This cleavage mechanism is dependent on factor of additional factors from those involved in base pairing. Thus, the specificity of action of a ribozyme may be greater than that of the antisense oligonucleotide binding the same RNA site. The enzyme ribozyme RNA molecule can be formed in a hammerhead motif, but it can also be formed in the motif of a hairpin, hepatitis delta virus, group I intron, or RNaseP-like RNA (in association with a RNA guide sequence). E-examples of such hammerhead motifs are described by Rossi (1992) Aids Research and Human Retroviruses 8:183; hairpin motifs by Hampel (1989) Biochemistry 28:4929, and Hampel (1990) Nuc. Acids Res. 18:299; the hepatitis delta virus motif by Perrotta (1992) Biochemistry 31:16; the RNaseP motif by Guerrier-Takada (1983) Cell 35:849; and the group I intron by Cech US Patent No. 4,987,071. The recitation of these specific motifs is not intended to be limiting; those skilled in the art will recognize that an enzymatic RNA molecule of this invention has a specific substrate binding site complementary to one or more of the target gene RNA regions, and has nucleotide sequence within or surrounding that substrate binding site that communicates an RNA-cleaving activity to the molecule. RNA interference (RNAi) In one aspect, the invention provides an RNA inhibitory molecule, a so-called "RNAi" molecule, comprising a phospholipase sequence of the invention. The RNAi molecule comprises a double-stranded RNA (dsRNA) molecule. RNAi can inhibit expression of a phospholipase gene. In one aspect, the RNAi is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more duplex nucleotides in length. While the invention is not limited by any particular mechanism of action, RNAi can enter a cell and cause the degradation of single-stranded RNA (ssRNA) of similar or identical sequences, including endogenous mRNAs. When a cell is exposed to double-stranded RNA (dsRNA), mRNA from the homologous gene is selectively graded by a process called RNA interference (RNAi). A possible basic mechanism behind RNAi is the breakdown of a double-stranded RNA (dsRNA) comparing a specific gene sequence into short pieces called short interfering RNA, which triggers the degradation of mRNA that matches its sequence. In one aspect, the RNAi's of the invention are employed in gene silencing therapeutics, see, for example, Shuey (2002) Drug Discov. Today 7:1040-1046. In one aspect, the invention provides methods for selectively degrading RNA employing the RNAi's of the invention. The process can be practiced in vitro, ex vivo or in vivo. In one aspect, the RNAi molecules of the invention can be employed to generate a loss-of-function mutation in a cell, an organ or an animal. Methods for preparing and using RNAi molecules to selectively degrade RNA are well known in the art, see, for example, U.S. Patent Nos. oS 6,506,559, 6,511,824, 6,515,109, 6,489,127. Nucleic Acid Modification The invention provides methods of generating variants of the nucleic acids of the invention, for example those encoding a phospholipase enzyme. In an alternative embodiment, the invention provides methods for modifying an enzyme of the invention, for example, by mutating its coding sequence by stochastic or random, or, non-stochastic, or "directed evolution" methods, such as Gene Site Saturation Mutagenesis® (GSSM®), to change the pH range of activity enzymes or optimal activity range, the temperature range of activity or optimal activity range, specificity, activity (kinetics); the use of enzyme glycosylation, phosphorylation, or metals (eg, Ca, Mg, Zn, Fe, Na), for example, to impact pH / temperature stability. The invention provides methods for modifying an enzyme of the invention, for example by mutating its coding sequence, for example by GSSM®, to increase its resistance to protease activity. The invention provides methods for modifying an enzyme of the invention, for example by mutating its coding sequence, for example by GSSM®, to modify the use of the enzyme from specific metal chelators for Ca, Mg, Na that would not chelate Zn. The invention provides methods for modifying an enzyme of the invention, for example by mutating its coding sequence, for example by GSSM® that would have a desired combination of activities, for example, PI, PA and PC / PE specific PLCs. These methods can be repeated or employed in various combinations to generate phospholipase enzymes having an altered or different activity or an altered or different stability than that of a standard nucleic acid-encoded phospholipase. These methods may likewise be repeated or employed in various combinations, for example, to generate variations in gene / message expression, message translation or message stability. In another aspect, the genetic makeup of a cell is altered by, for example, modification of a homologous gene ex vivo, followed by its reinsertion into the cell. A nucleic acid of the invention can be altered by any means. For example, stochastic or random methods, or, non-stochastic, or "directed evolution" methods. Methods for random mutation of genes are well known in the art, see, for example, U.S. Patent. At the. 5,830,696. For example, mutagens can be used to randomly mutate a gene. Mutagens include, for example, ultraviolet light or gamma irradiation, or a chemical mutagen, for example, mitomycin, nitrous acid, photoactivated psoralens, alone or in combination, to induce mild DNA breaks to repair by recombination. Other chemical mutagens include, for example, sodium bisulfite, nitrous acid, hydroxylamine, hydrazine or formic acid. Other mutagens are nucleotide precursor analogues, eg nitrosoguanidine, 5-bromouracil, 2-aminopurine or acridine. These agents can be added in a PCR reaction in place of the nucleotide precursor thereby mutating the sequence. Intercalating agents such as proflavine, acriflavine, quinacrine and the like may likewise be employed. Any technique in molecular biology can be used, for eg random PCR mutagenesis, see eg Rice (1992) Proc. natl. academy Sci. USA 89:5467-5471; or, combinatorial multiple cassette mutagenesis, see, for example, Crameri (1995) Biotechniques 18:194-196. Alternatively, nucleic acids, eg genes, can be reassembled after random, or "stochastic" fragmentation, see, for example, US Patents no. oS 6,291,242, 6,287,862, 6,287,861, 5,955,358, 5,830,721, 5,824,514, 5,811,238, 5,605,793. In alternative aspects, modifications, additions or deletions are introduced by error-prone PCR, scrambling, oligonucleotide-directed mutagenesis, cluster PCR, sexual PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive ensemble mutagenesis, exponential ensemble mutagenesis , site-specific mutagenesis, gene reassortment, Gene Site Saturation Mutagenesis® (GSSM®), synthetic linkage reassortment (SLR), recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, uracil-containing standard mutagenesis, spaced duplex mutagenesis, point mismatch repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiogenic mutagenesis, deletion mutagenesis, strict selection mutagenesis, strict purification mutagenesis, artificial gene synthesis, ensemble mutagenesis, breeding of chimeric nucleic acid multimer, and / or a combination thereof and similar methods. The following publications describe a variety of recursive recombination methods and / or procedures that can be incorporated into the methods of the invention: Stemmer (1999) "Molecular breeding of viruses for targeting and other clinical properties" Tumor Targeting 4:1-4; Ness (1999) Nature Biotechnology 17:893 - 896; Chang (1999) "Evolution of a cytokine using DNA family shuffling" Nature Biotechnology 17:793 - 797; Min-shull (1999) "Protein evolution by molecular breeding" Current Opinion in Chemical Biology 3:284 - 290; Christians (1999) "Directed evolution of thymidine kinase for AZT phosphorylation using DNA family shuffling" Nature Biotechnology 17:259 - 264; Crameri (1998) "DNA shuffling of a family of genes from diverse species accelerates directed evolution" Nature 391:288-291; Crameri (1997) "Molecular evolution of an arsenate detoxification pathway by DNA shuffling", Nature Biotechnology 15:436 - 438; Zhang (1997) "Directed evolution of an effective fucosidase from a galactosidase by DNA shuffling and screening" Proc. natl. academy Sci. USA 94:4504-4509; Patten et al. (1997) "Applications of DNA Shuffling to Pharmaceuticals and Vaccines" Current Opinion in Biotechnology 8:724 - 733 ; Crameri et al. (1996) "Construction and evolution of antibody-phage libraries by DNA shuffling" Nature Medicine 2:100-103; Crameri et al. (1996) "Improved green fluorescent protein by molecular evolution using DNA shuffling" Nature Biotechnology 14:315 - 319; Gates et al. (1996) "Affinity selective isolation of ligands from peptide libraries through display on a lac repressor 'headpiece dimer'" Journal of Molecular Biology 255:373-386; Stemmer (1996) "Sexual PCR and Assembly PCR" In: The Encyclopedia of Molecular Biology. VCH Publishers, New York. pp. 447 - 457; Crameri and Stemmer (1995) "Combinatorial multiple cassette mutagenesis creates all the permutations of mutant and wildtype cassettes" BioTechniques 18:194 - 195; Stemmer et al. (1995) "Single-step assembly of a gene and entire plasmid form large numbers of oligodeoxyribonucleotides" Gene, 164:49-53; Stemmer (1995) "The Evolution of Molecular Computation" Science 270: 1510; Stemmer (1995) "Searching Sequence Space" Bio / Technology 13:549 - 553; Stemmer (1994) "Rapid evolution of a protein in vitro by DNA shuffling" Nature 370:389 - 391; and Stemmer (1994) "DNA shuffling by random fragmentation and reassembly: In vitro recombination for molecular evolution." process natl. academy Sci. USA 91:10747-10751. Mutational methods of generating diversity include, for example, site-directed mutagenesis (Ling et al. (1997) "Approaches to DNA mutagenesis: an overview" Anal Biochem. 254(2): 157-178; Dale et al. (1996) ) "Oligonucleotide-directed random mutagenesis using the phosphorothioate method" Methods Mol. Biol. 57:369 - 374; Smith (1985) "In vitro mutagenesis" Ann. Rev. Genet. 19:423-462; Botstein & Shortle (1985) "Strategies and applications of in vitro mutagenesis" Science 229:1193-1201; Carter (1986) "Site-directed mutagenesis" Biochem. J. 237:1-7; and Kunkel (1987) "The efficiency of oligonucleotide directed mutagenesis" in Nucleic Acids & Molecular Biology (Eckstein, F. and Lilley, D. M. J. eds., S-pringer Verlag, Berlin)); mutagenesis using uracil-containing patterns (Kunkel (1985) "Rapid and efficient site-specific mutagenesis without phe- 5 notypic selection" Proc. Natl. Acad. Sci. USA 82:488 - 492; Kunkel et al. (1987) "Rapid and efficient site-specific mutagenesis without phenotypic selection" Methods in Enzymol. 154, 367-382; and Bass et al. similar (1988) "Mutant Trp repressors with new DNA-binding specificities" Science 242:240 - 245); oligonucleotide-directed mutagenesis (Methods in Enzymol. 100: 10 468 - 500 (1983); Methods in Enzymol. 154: 329 - 350 (1987); Zoller & Smith (1982) "Oligonucleotide - directed mutagenesis using M13 - derived vectors: an efficient and general procedure for the production of point mutations in any DNA fragment" Nucleic Acids Res. 10:6487 - 6500; Zoller & Smith (1983) "Oligonucleotide - directed mutagenesis of DNA fragments cloned into M13 15 vectors" Methods in Enzymol. 100:468 - 500; and Zoller & Smith (1987) "Oli gonucleotide - directed mutagenesis: a simple method using two oligonucleotide primers and a single - stranded DNA template" Methods in Enzymol. 154:329 - 350); mutagenesis of phosphothioate-modified DNA (Taylor et al. (1985) "The use of phosphorothioate - modified DNA in restriction 20 enzyme reactions to prepare nicked DNA" NucL Acids Res. 13: 8749 - 8764; Taylor et al. (1985) "The rapid generation of oligonucleotide - directed mutations at high frequency using phosphorothioate - modified DNA" NucL Acids Res. 13: 8765 - 8787 (1985); Nakamaye (1986) "Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application 25 to oligonucleotide - directed mutagenesis" NucL Acids Res. 14: 9679 - 9698; Sayers et al. (1988) "Y - T Exonucleases in phosphorothioate - based oligonucleotide - directed mutagenesis" NucL Acids Res. 16:791 - 802; and Sayers et al. (1988) "Strand specific cleavage of phosphorothioate - containing DNA by reaction with restriction endonucleases in the presence of 30 ethidium bromide” NucL Acids Res. 16: 803 - 814); generating duplex DNA with space (Kramer et al. (1984) "The gapped duplex DNA approach to oligonucleotide - directed mutation construction” Nucl. Acids Res. 12: 9441 - 9456; Kramer & Fritz (1987) Methods in Enzy-mol. "Oligonucleotide - directed construction of mutations via gapped duplex DNA" 154:350 - 367; Kramer et al. (1988) "Improved enzymatic in vitro reactions in the gapped duplex DNA approach to oligonucleotide - directed construction of mutations" Nucl. Acids Res. 16: 7207; and Fritz et al. (1988) "Oligonucleotide - directed construction of mutations: a gapped duplex DNA procedure without enzymatic reactions in vitro" Nucl. Acids Res. 16: 6987 - 6999). Additional protocols employed in the methods of the invention include point mismatch repair (Kramer (1984) "Point Mismatch Repair" Cell 38:879 - 887), mutagenesis employing repair-deficient host strains (Carter et al. (1985) "Improved oligonucleotide site - directed mutagenesis using M13 vectors" Nucl. Acids Res. 13: 4431 - 4443; and Carter (1987) "Improved oligonucleotide - directed mutagenesis using M13 vectors" Methods in Enzymol. 154: 382 - 403), deletion mutagenesis ( Eghtedarzadeh (1986) "Use of oligonucleotides to generate large deletions" Nucl. Acids Res. 14: 5115), restricted-selection and restricted-selection and restricted-purification (Wells et al. (1986) "Importance of hydrogen - bond formation in stabilizing the transition state of subtilisin'' Phil. Trans. R. Soc. Lond. A 317: 415 - 423), mutagenesis by total gene synthesis (Nambiar et al. (1984) "Total synthesis and cloning of a gene coding for the ribonuclease S protein" Science 223: 1299 - 1301; Sakamar and Khorana (1988) "Total synthesis and expression of a gene for the a - subunit of bovine rod outer segment guanine nucleotide - binding protein (trans-ducin)" Nucl. Acids Res. 14: 6361 - 6372; Wells et al. (1985) "Cassette mutagenesis: an efficient method for generation of multiple mutations at defined sites" Gene 34:315 - 323; and Grundstrom et al. (1985) "Oligonucleotide - directed mutagenesis by microscale 'shot - gun' gene synthesis" Nucl. Acids Res. 13: 3305 - 3316), double strand break repair (Mandecki (1986); Arnold (1993) "Protein engineering for unusual environments" Current Opinion in Biotechnology 4:450 - 455. "Oligonucleotide - directed double - strand break repair in plasmids of Escherichia coli: a method for site - specific mutagenesis" Proc. Natl. Acad. Sci. USA, 83:7177 - 7181). Additional details on many of the above methods can be found in Methods in Enzymology Volume 154, which likewise describes useful controls for site-specific mutagenesis. diagnose and troubleshoot various mutagenesis methods. See also United States Patent No. 5,605,793 to Stemmer (February 25, 1997), "Methods for In Vitro Recombination"; United States Patent No. 5,811,238 to Stemmer and the like (September 22, 1998) "Methods for Generating Polynucleotides having Desired Characteristics by Iterative Selection and Recombination"; United States Patent No. 5,830,721 to Stemmer and the like (November 3, 1998), "DNA Mutagenesis by Random Fragmentation and Reassembly"; U.S. Patent At the. 5,834,252 to Stemmer, and the like (Nov. 10, 1998) "End - Complementary Polymerase Reaction"; United States Patent No. 5,837,458 to Minshull, et al. (November 17, 1998), "Methods and Compositions for Cellular and Metabolic Engineering"; WO 95 / 22625, Stemmer and Crameri, "Mutagenesis by Random Fragmentation and Reassembly"; WO 96 / 33207 by Stemmer and Lipschütz "End Complementary Polymerase Chain Reaction"; WO 97 / 20078 by Stemmer and Crameri "Methods for Generating Polynucleotides having Desired Characteristics by Iterative Selection and Recombination"; WO 97 / 35966 by Minshull and Stemmer, "Methods and Compositions for Cellular and Metabolic Engineering"; WO 99 / 41402 by Punnonen and the like "Targeting of Genetic Vaccine Vectors"; WO 99 / 41383 by Punnonen and the like "Antigen Library Immunization"; WO 99 / 41369 by Punnonen and the like "Genetic Vaccine Vector Engineering"; WO 99 / 41368 by Punnonen and the like "Optimization of Immunomodulatory Properties of Genetic Vaccines"; EP 752008 by Stemmer and Crameri, "DNA Mutagenesis by Random Fragmentation and Reassembly"; EP 0932670 by Stemmer "Evolving Cellular DNA Uptake by Recursive Sequence Recombination"; WO 99 / 23107 by Stemmer and the like, "Modification of Virus Tropism and Host Range by Viral Genome Shuffling"; WO 99 / 21979 by Apt and the like, "Human Papillomavirus Vectors"; WO 98 / 31837 by del Cardayre and the like "Evolution of Whole Cells and Organisms by Recursive Sequence Recombination"; WO 98 / 27230 by Patten and Stemmer, "Methods and Compositions for Polypeptide Engineering"; WO 98 / 27230 by Stemmer and the like, "Methods for Optimization of Gene Therapy by Recursive Sequence Shuffling and Selection", WO 00 / 00632, "Methods for Generating Highly Diverse Libraries", WO 00 / 09679, "Methods for Obtaining in Vitro Recombined Polynucleotide Sequence Banks and Resulting Sequences", WO 98 / 42832 by Arnold and the like, "Recombination of Polynucleotide Sequences Using Random or Defined Primers", WO 99 / 29902 by Arnold and the like, "Method for Creating Polynucleotide and Polypeptide Sequences", WO 98 / 41653 by Vind, "An in vitro Method for Construction of a DNA Library", WO 98 / 41622 by Borchert and the like, "Method for Constructing a Library Using DNA Shuffling", and WO 98 / 42727 by Pati and Zarling, "Sequence Alterations using Homologous Recombination". Certain U.S. orders provide additional details with respect to the various diversity generation methods, including "SHUFFLING OF CODON ALTERED GENES" by Patten and the like filed September 28, 1999, (U.S. Ser. No. 09 / 407,800); "EVOLUTION OF WHOLE CELLS AND ORGANISM BY RECURSIVE SEQUENCE RECOMBINATION" by del Cardayre and the like, filed July 15, 1998 (U.S. Ser. No. 09 / 166,188), and July 15, 1999 (U.S. Ser. No. 09 / 354,922); "OLIGONUCLEOTIDE MEDIATED NUCLEIC ACID RECOMBINATION" by Crameri and the like, filed September 28, 1999 (U.S. Ser. No. 09 / 408,392), and "OLIGONUCLEOTIDE MEDIATED NUCLEIC ACID RECOMBINATION" by Crameri and the like, filed January 18, 2000 (PCT / USOO / 01203); "USE OF CODON - VARIED OLIGONUCLEOTIDE SYNTHESIS FOR SYNTHETIC SHUFFLING" by Welch and the like, filed September 28, 1999 (U.S. Ser. No. 09 / 408,393); "METHODS FOR MAKING CHARACTER STRINGS, POLYNUCLEOTIDES & POLYPEPTIDES HAVING DESIRED CHARACTERISTICS" by Selifonov and the like, filed January 18, 2000, (PCT / US00 / 01202) and, for example, "METHODS FOR MAKING CHARACTER STRINGS, POLYNUCLEOTIDES & POLYPEPTIDES HAVING DESIRED CHARACTERISTICS" by Selifonov and the like, filed July 18, 2000 (U.S. Ser. No. 09 / 618,579); "METHODS OF POPULATING DATA STRUCTURES FOR USE IN EVOLUTIONARY SIMULATIONS" by Selifonov and Stemmer, filed July 18 January, 2000 (PCT / US00 / 01138); and "SINGLE - STRANDED NUCLEIC ACID TEMPLATE - MEDIATED RECOMBINATION AND NUCLEIC ACID FRAGMENT ISOLATION" by Affholter, filed September 6, 2000 (U.S. Ser. No. 09 / 656,549). Non-stochastic or "directed evolution" methods including, for example, saturation mutagenesis (e.g. GSSM®), synthetic linkage reassortment (SLR), or a combination thereof, are employed to modify the nucleic acids of the invention. to generate phospholipases with new or altered properties (eg, activity under highly acidic or alkaline conditions, high temperatures, and the like). Polypeptides encoded by the modified nucleic acids can be evaluated for an activity before testing for a phospholipase or other activity. Any test modality or protocol can be used, for example, using a capillary array platform. See, for example, US Patents No. oS 6,280,926 and 5,939,250. Saturation Mutagenesis, or, GSSM® In one aspect of the invention, non-stochastic gene modification, a "directed evolution process", is employed to generate phospholipases with new or altered properties. Variations of this method were called "gene site mutagenesis", "site saturation mutagenesis", "Gene site saturation mutagenesis®" or simply "GSSM®". They can be used in combination with other mutagenization processes. See, for example, US Patents No. oS 6,171,820 and 6,238,884. In one aspect, GSSM® comprises providing a standard polynucleotide and a plurality of oligonucleotides, wherein each oligonucleotide comprises a sequence homologous to the standard polynucleotide, whereby to target a specific sequence of the standard polynucleotide, and a sequence that is a variant of the homologous gene; generate pollu- Progeny cleotides comprise non-stochastic sequence variations by replicating the standard polynucleotide with the oligonucleotides, whereby generating polynucleotides comprises homologous gene sequence variations. In one aspect, codon primers containing a degenerate N,N,G / T sequence are employed to introduce point mutations into a polynucleotide to generate a group of progeny polypeptides in which a full range of single amino acid substitutions is represented in each amino acid position, for example, an amino acid residue at an enzyme active site or binding site of the targeted ligand to be modified. These oligonucleotides may comprise a first contiguous homologous sequence, a degenerate N,N,G / T sequence, and, optionally, a second homologous sequence. Downstream translational products from the use of such oligonucleotides include all possible amino acid changes at each amino acid site along the polypeptide, because the degeneracy of the N,N,G / T sequence includes codons for all 20 amino acids. . In one aspect, such a degenerate oligonucleotide (comprised of, for example, a degenerate N,N,G / T cassette) is employed to subject each original codon in a parental polynucleotide pattern over a full range of codon substitutions. . In another aspect, at least two degenerate cassettes are employed - either in the same oligonucleotide or not, by submitting at least two original codons in a parental polynucleotide pattern over a full range of codon substitutions. For example, more than one N,N,G / T sequence may be contained in an oligonucleotide to introduce amino acid mutations at more than one site. This plurality of N,N,G / T sequences may be directly contiguous, or separated by one or more additional nucleotide sequence(s). In another aspect, oligonucleotides useful for introducing additions and deletions may be employed alone or in combination with codons containing an N,N,G / T sequence, to introduce any combination or permutation of amino acid substitutions, deletions and / or additions. . In one aspect, simultaneous mutagenesis of two or more contiguous amino acid positions is accomplished by employing an oligonucleotide that contains contiguous N,N,G / T triplets, that is, a sequence of (N,N,G / T)n degenerate. In another aspect, degenerate cassettes having less degeneracy than the N,N,G / T sequence are employed. For example, it may be desirable in some examples to use (for example in an oligonucleotide) a degenerate triplet sequence comprised of only one N where said N may be at the first, second or third position of the triplet. Any other bases that include any combinations and permutations thereof may be employed in the remaining two positions of the triplet. Alternatively, it may be desirable in some examples to use (for example in an oligo) a degenerate N,N,N triplet sequence. In one aspect, the use of degenerate triplets (e.g., N,N,G / T triplets) allows for the systematic and easy generation of a full range of possible natural amino acids (for a total of 20 amino acids) in each and every amino acid positions in a polypeptide (in alternative aspects, methods likewise include generating less than all possible substitutions per amino acid residue, or codon, position). For example, for a 100 amino acid polypeptide, 2000 distinct species (ie, 20 possible amino acids per position X 100 amino acid positions) can be generated. By using an oligonucleotide or group of oligonucleotides containing a degenerate N,N,G / T triplet, 32 individual sequences can code for all 20 possible natural amino acids. Thus, in a reaction vessel in which a parental polynucleotide sequence is subjected to saturation mutagenesis using at least one such oligonucleotide, 32 distinct progeny polynucleotides encoding 20 distinct polypeptides are generated. In contrast, the use of a non-degenerate oligonucleotide in site-directed mutagenesis leads to only one progeny polypeptide product per reaction vessel. Non-degenerate oligonucleotides may optionally be employed in combination with the degenerate primers described; per For example, non-degenerate oligonucleotides can be used to generate specific point mutations in a splice polynucleotide. This provides a means of generating specific silent point mutations, point mutations that lead to corresponding amino acid changes, and point mutations that cause the generation of stop codons and the corresponding expression of polypeptide fragments. In one aspect, each saturation mutagenesis reaction vessel contains polynucleotides that encode at least 20 progeny polypeptide molecules (e.g., phospholipase) such that all 20 naturally occurring amino acids are represented at a specific amino acid position corresponding to the position of codon mutagenized in the parent polynucleotide (other aspects use less than all 20 natural combinations). The degenerate descendant polypeptides of 32 duplications generated from each saturation mutagenesis reaction vessel can be subjected to clonal amplification (e.g., cloned from into a suitable host, e.g. E. coli host, using, for example, example, an expression vector) and subjected to expression evaluation. When an individual progeny polypeptide is identified by evaluating itself to exhibit a favorable change in property (when compared to the parent polypeptide, such as increased phospholipase activity under alkaline or acidic conditions), it can be sequenced to identify the correspondingly favorable amino acid substitution contained therein. . In one aspect, in mutagenizing each and all amino acid positions in a parent polypeptide employing saturation mutagenesis as described herein, favorable amino acid changes can be identified at more than one amino acid position. One or more new progeny molecules can be generated which contain a combination of all or part of these favorable amino acid substitutions. For example, if 2 specific favorable amino acid changes are identified at each of the 3 amino acid positions in a polypeptide, the permutations include 3 possibilities at each position (no change from the original amino acid, and each of the two changes favorable rations) and 3 positions. In this way, there are 3 x 3 x 3 or 27 total possibilities, including 7 that were previously examined - 6 single point mutations (ie, 2 at each of the three positions) and no change at any position. In another aspect, site saturation mutagenesis may be employed together with other stochastic or non-stochastic means to vary the sequence, e.g. synthetic linkage reassortment (see below), scrambling, chimerization, recombination and similar mutagenization processes and decontamination agents. mutagenization. This invention provides the use of any(any) mutagenization process(es), including saturation mutagenesis, in an iterative manner. Synthetic Link Regrouping (SLR) The invention provides a non-stochastic gene modification system called "synthetic linkage reassembly", or simply "SLR", a "directed evolution process", to generate phospholipases with new or altered properties. SLR is a method of linking oligonucleotide fragments together non-stochastically. This method differs from stochastic oligonucleotide shuffling in that the nucleic acid building blocks are not randomly shuffled, concatenated or chimerized, but rather are non-stochastically clustered. See, for example, U.S. Patent Application. At the. Serial (USSN) 09 / 332,835 entitled "Synthetic Ligation Reassembly in Directed Evolution" and filed June 14, 1999 ("USSN 09 / 332,835"). In one aspect, SLR comprises the following steps: (a) providing a standard polynucleotide, wherein the standard polynucleotide comprises sequence encoding a homologous gene; (b) providing a plurality of building block polynucleotides, wherein the building block polynucleotides are intended to reassemble by crossover with the standard polynucleotide in a predetermined sequence, and a building block polynucleotide comprises a sequence that is a variant the homologous gene and a sequence homologous to the standard polynucleotide flanking the variant sequence; (c) combine a building block polynucleotide with a standard polynucleotide such that the building block polynucleotide assembles by crossing over with the standard polynucleotide to generate polynucleotides comprising homologous gene sequence variations. SLR does not depend on the presence of high levels of homology between polynucleotides to be reassembled. In this way, this method can be used to non-stochastically generate libraries (or groups) of progeny molecules comprised of more than 10 100 different chimeras. SLR can be used to generate libraries comprised of more than 10 1000 different descendant chimeras. Thus, aspects of the present invention include non-stochastic methods of producing a finished chimeric nucleic acid molecule group by scraping a total grouping order that is selected by design. This method includes the steps of generating by design a plurality of specific nucleic acid building blocks having useful mutually compatible linkable ends, and assembling these nucleic acid building blocks such that a designated total grouping order is obtained. The mutually compatible ligatable ends of the nucleic acid building blocks to be grouped are considered "useful" for this type of ordered grouping, if they allow the building blocks to be grouped in predetermined orders. In this way, the total grouping order in which the nucleic acid building blocks can be coupled is specified by the design of the pluggable ends. If more than one clustering step is to be employed, then the total clustering order in which the nucleic acid building blocks can be clustered is likewise specified by the sequential order of clustering step(s). In one aspect, the annealed building pieces are treated with an enzyme, such as a ligase (e.g. T4 DNA ligase), to covalently link the building pieces. In one aspect, the design of oligonucleotide building blocks is achieved by analyzing a group of acidic sequence patterns. from the parent nucleotide that serves as a base to produce a progeny of finished chimeric polynucleotides. These parental oligonucleotide patterns in this way serve as a source of sequence information that aid in the design of the nucleic acid building blocks that are to be mutagenized, eg chimerized or scrambled. In one aspect of this method, the sequences of a plurality of parental nucleic acid patterns are aligned in order to select one or more demarcation points. Demarcation points can be located in an area of ​​homology, and are comprised of one or more nucleotides. These demarcation points are preferably shared by at least two of the parent patterns. Demarcation points can be used to delineate the boundaries of the oligonucleotide building blocks to be generated in order to reassemble the parent polynucleotides. The identified and selected demarcation points on the parent molecules serve as potential chimerization points in the clustering of the final chimeric progeny molecules. A demarcation point can be an area of ​​homology (comprised of at least one homologous nucleotide base) shared by at least two parental polynucleotide sequences. Alternatively, a demarcation point may be an area of ​​homology that is shared by at least half of the parental polynucleotide sequences, or, it may be an area of ​​homology that is shared by at least two-thirds of the parental polynucleotide sequences. Even more preferably useful demarcation points is an area of ​​homology that is shared by at least three quarters of the parental polynucleotide sequences, or, may be shared by nearly all of the parental polynucleotide sequences. In one aspect, a demarcation point is an area of ​​homology that is shared by all parental polynucleotide sequences. In one aspect, a linkage regrouping process is performed exhaustively in order to generate an exhaustive library of polynucleotides. descendant chimeric cleotids. In other words, all possible ordered combinations of the nucleic acid building blocks are represented in the group of finished chimeric nucleic acid molecules. At the same time, in another embodiment, the clustering order (i.e., the clustering order of each building block in the 5' to 3 sequence of each finished chimeric nucleic acid) in each combination is by design (or non-stochastic). ) as described above. Because of the non-stochastic nature of this invention, the possibility of unwanted side products is greatly reduced. 10 In another aspect, the linkage regrouping method is real systematically. For example, the method is carried out in order to generate a systematically compartmentalized library of progeny molecules, with compartments that can be systematically evaluated, for example one by one. In other words, this invention provides that, 15 Through the selective and judicious use of specific nucleic acid building blocks, together with the selective and judicious use of sequentially staggered clustering reactions, a design can be obtained where specific clusters of descendant products are made in each of several vessels of reaction. This allows a systematic examination and evaluation procedure to be carried out. In this way, these methods allow potentially very large numbers of progeny molecules to be examined systematically in smaller groups. Because of their ability to perform chimerizations in a way that is highly flexible, yet equally exhaustive and systematic, particularly when there is a low level of homology between the parent molecules, these methods provide for the generation of a library (or group) understood of a large number of descending molecules. Because of the non-stochastic nature of the binding reassortment present invention, the offspring molecules generated preferably comprise a library of finalized chimeric nucleic acid molecules 30 molecules having a total grouping order that is selected by design. Saturation mutagenesis and similarly optimized directed evolution methods can be employed. nailed together to generate different descendant molecular species. It is appreciated that the invention provides freedom of choice and control with respect to the selection of demarcation points, the size and number of nucleic acid building blocks, and the size and design of couplings. Furthermore, it is appreciated that the requirement for intermolecular homology is highly relaxed as to the operability of this invention. In fact, demarcation points can still be selected in areas of little or no intermolecular homology. For example, because of codon hydrogen bond variations, that is, codon degeneracy, nucleotide substitutions can be introduced into nucleic acid building blocks without altering the amino acid originally encoded in the corresponding parent pattern. Alternatively, a codon can be changed such that the coding for an originally amino acid is changed. This invention provides that such substitutions can be introduced into the nucleic acid building block in order to increase the incidence of intermolecularly homologous demarcation points and in this way allow an increased number of couplings to be obtained between the building blocks, which successively allow for a greater number of descending chimeric molecules will be generated. In another aspect, the synthetic nature of the step in which the building blocks are generated allows for the design and introduction of nucleotides (e.g. one or more nucleotides, which may be, for example, codons or introns or regulatory sequences) which may then optionally be removed in an in vitro process (e.g. by mutagenesis) or in an in vivo process (e.g. using the gene splicing capability of a host organism). It is appreciated that in many instances the introduction of these nucleotides may likewise be desirable for many reasons other than the potential benefit of creating a useful demarcation point. In one aspect, a nucleic acid building block is employed to introduce an intron. In this way, functional introns are introduced into an artificial gene manufactured according to the methods described here. The artificially introduced intron(s) may be functional in a host cell for gene splicing in approximately the same way that naturally occurring introns functionally serve in gene splicing. Optimized Directed Evolution System The invention provides a non-stochastic gene modification system called "optimized directed evolution system" to generate phospholipases with new or altered properties. Optimized directed evolution is directed to the use of repeated cycles of reductive reclassification, recombination and selection that allow the directed molecular evolution of nucleic acids through recombination. Optimized directed evolution allows the generation of a large population of evolved chimeric sequences, where the generated population is significantly enriched for sequences that have a predetermined number of crossover events. A crossover event is a point in a chimeric sequence where a change in sequence occurs from one parental variant to another parental variant. Such a point is usually at the junction where oligonucleotides from two sources are linked together to form a single sequence. This method allows the calculation of the correct concentrations of oligonucleotide sequences so that the final chimeric population of sequences is enriched for the selected number of crossover events. This provides more control over the choice of chimeric variants that have a predetermined number of crossover events. Furthermore, this method provides a convenient means to explore a tremendous amount of possible protein variant space compared to other systems. Previously, if someone generated, for example, 10 13 chimeric molecules during a reaction, it would be extremely difficult to test such a high number of chimeric variants for a particular activity. Furthermore, a significant portion of the offspring population would have a very high number of crossover events that would result in proteins, which are less likely to have increased levels of an activity. private life. Using these methods, the population of chimeric molecules can be enriched for those variants that have a particular number of crossover events. In this way, although one can still generate 10 13 chimeric molecules during a reaction, each of the 5 molecules selected for further analysis most likely have, for example, only three crossover events. Because the resulting offspring population may be prone to having a predetermined number of crossover events, the limits on functional variety between chimeric molecules are reduced. This provides a more manageable number of variables when calculating which oligonucleotide from the original parental polynucleotides could be responsible for affecting a particular trait. One method of creating a chimeric progeny polynucleotide sequence is to create oligonucleotides corresponding to fragments or portions of each parent sequence. Each oligonucleotide preferably includes a single region of overlap so that mixing the oligonucleotides together results in a new variant that has each oligonucleotide fragment assembled in the correct order. Additional information can also be found in USSN 09 / 332,835. The number of oligonucleotides generated for each parental variant is linked to the total number of crosses resulting in the chimeric molecule that is ultimately created. For example, three parental nucleotide sequence variants could be provided to undergo a ligation reaction in order to find a chimeric variant having, for example, greater activity at high temperature. As an example, a group of 50-25 oligonucleotide sequences can be generated corresponding to each of the portions of each parental variant. Consequently, during the ligation reassembly process there could be up to 50 crossover events within each of the chimeric sequences. The probability that each of the generated chimeric polynucleotides will contain oligonucleotides of each parental variant in alternating order is very low. If each oligonucleotide fragment is present in the ligation reaction in the same molar amount, it is likely that at some positions the oligonucleotides tides from the same parental polynucleotide will then bind to each other and thus will not result in a crossover event. If the concentration of each oligonucleotide from each origin is held constant during any binding step in this example, there is a 1 / 3 chance (assuming 3 origins) that an oligonucleotide of the same parental variant will bind within the chimeric sequence and not produce crossover. . Consequently, a probability density function (PDF) can be determined to predict the population of crossover events that are likely to occur during each step in a ligation reaction, given a fixed number of parental variants, several oligonucleotides that correspond to each variant, and the concentrations of each variant during each step in the ligation reaction. The statistics and math after determining the PDF are described below. Using these methods, one can calculate such a probability density function, and in this way enrich the chimeric progeny population for a predetermined number of crossover events that are the result of a particular binding reaction. In addition, a target number of crossover events can be predetermined, and the system then programmed to calculate the starting amounts of each parent oligonucleotide during each step in the ligation reaction to result in a probability density function that centers the predetermined number of crossover events. These methods are directed at using repeated cycles of reductive reclassification, recombination and selection that allow for the directed molecular evolution of a nucleic acid encoding a polypeptide through recombination. This system allows for the generation of a large population of evolved chimeric sequences, where the generated population is significantly enriched for sequences that have a predetermined number of crossover events. A crossover event is a point in a chimeric sequence where a change in sequence occurs from one parental variant to another parental variant. Such a point is normally at the junction where oligonucleotides from two sources are linked together to form a single sequence. the month The whole allows calculation of the correct concentrations of oligonucleotide sequences so that the final chimeric population of sequences is enriched for the selected number of crossover events. This provides more control over choosing chimeric variants that have a predetermined number of crossover events. Furthermore, these methods provide a convenient means to explore the tremendous amount of protein variant space possible compared to other systems. Using the methods described herein, the population of chimeric molecules can be enriched for those variants that have a particular number of crossover events. In this way, although one can still generate 10 13 chimeric molecules during a reaction, each of the molecules selected for further analysis is more likely to have, for example, only three crossover events. Because the resulting offspring population may be prone to having a predetermined number of crossover events, the limits on functional variety between chimeric molecules are reduced. This provides a more manageable number of variables when calculating which oligonucleotide from the original parental polynucleotides could be responsible for affecting a particular trait. In one aspect, the method creates a chimeric progeny polynucleotide sequence by creating oligonucleotides that correspond to fragments or portions of each parent sequence. Each oligonucleotide preferably includes a single region of overlap so that mixing the oligonucleotides together results in a new variant that has each oligonucleotide fragment assembled in the correct order. Also see USSN 09 / 332,835. The number of oligonucleotides generated for each parental variant has a bearing on the total number of crosses resulting in the chimeric molecule that is ultimately created. For example, three parental nucleotide sequence variants could be provided to undergo a ligation reaction in order to find a chimeric variant having, for example, greater activity at high temperature. As an example, a group of 50 oligonucleotide sequences can be generated corresponding to each of the portions of each parental variant. Consequently, during the linkage reassembly process there could be up to 50 crossover events within each of the chimeric sequences. The probability that each of the generated chimeric polynucleotides will contain oligonucleotides from each parental variant in alternating order is very low. If each oligonucleotide fragment is present in the ligation reaction in the same molar amount, it is likely that at some positions, oligonucleotides from the same parent polynucleotide will ligate next to each other and thus not result in a crossover event. If the concentration of each oligonucleotide from each origin is held constant during any binding step in this example, there is a 1 / 3 chance (assuming 3 origins) that an oligonucleotide of the same parental variant will bind within the chimeric sequence and not produce crossover. . Consequently, a probability density function (PDF) can be determined to predict the population of crossover events that are likely to occur during each step in a ligation reaction, given a fixed number of parental variants, several oligonucleotides that correspond to each variant, and the concentrations of each variant during each step in the ligation reaction. The statistics and math after determining the PDF are described below. One can calculate such a probability density function, and in this way enrich the chimeric offspring population for a predetermined number of crossover events that are the result of a particular binding reaction. In addition, a target number of crossover events can be predetermined, and the system then programmed to calculate the starting amounts of each parent oligonucleotide during each step in the ligation reaction to result in a probability density function that centers the predetermined number of crossover events. Determining Crossing Events Embodiments of the invention include a system and software that receive a desired crossover probability density (PDF) function, the number of source genes to be reassembled, and the number of fragments in the reassemble as consumptions. The output of this program is a "fragment PDF" that can be employed to determine a recipe for producing reassorted genes, and the estimated crossover PDF of those genes. The process described herein is preferably performed in MATLAB® (The Mathworks, Natick, Massachusetts) a programming language and development environment for technical computing. Iterative Processes In practicing the invention, these processes can be iteratively repeated. For example, a nucleic acid (or, the nucleic acid) responsible for an altered phospholipase phenotype is identified, re-isolated, re-modified, retested for activity. This process can be iteratively repeated until a desired phenotype is constructed. For example, an entire biochemical anabolic or catabolic pathway can be built up in a cell, including phospholipase activity. Similarly, if it is determined that a particular oligonucleotide does not affect in any way the desired trait (e.g., a new phospholipase phenotype), it can be removed as a variable by synthesizing larger parent oligonucleotides that include the sequence to be removed. Since the incorporation of the sequence into a larger sequence prevents any crossover events, there will no longer be any variation of this sequence in the descendant polynucleotides. This iterative practice of determining which oligonucleotides are most related to the desired trait, and which are unrelated, allows for more efficient exploration of all possible protein variants that could be provided for a particular activity or trait. in vivo shuffling In vivo shuffling of molecules is employed in methods of the invention that provide variants of polypeptides of the invention, for example, antibodies, phospholipase enzymes, and the like. In vivo shuffling can be performed using the natural property of cells to re- combine multimers. While in vivo recombination has provided the main natural routine for molecular diversity, genetic recombination remains a relatively complex process that involves 1) the recognition of homologies; 2) filament cleavage, filament invasion, and metabolic steps leading to the production of recombinant chiasm; and finally 3) the resolution of chiasm in discrete recombination molecules. Chiasm formation requires recognition of homologous sequences. In one aspect, the invention provides a method for producing a hybrid polynucleotide from at least a first polynucleotide and a second polynucleotide. The invention can be employed to produce a hybrid polynucleotide by introducing at least a first polynucleotide and a second polynucleotide that share at least a region of partial sequence homology into a suitable host cell. Partial sequence homology regions promote processes that result in sequence reorganization producing a hybrid polynucleotide. The term "hybrid polynucleotide", as used herein, is any nucleotide sequence that results from the method of the present invention and contains a sequence of at least two original polynucleotide sequences. Such hybrid polynucleotides can result from intermolecular recombination events that promote sequence integration between DNA molecules. Furthermore, such hybrid polynucleotides can result from intramolecular reductive reclassification processes that use repeated sequences to alter a nucleotide sequence within a DNA molecule. Production of sequence variants The invention also provides methods of preparing nucleic acid sequence variants and phospholipase sequences of the invention or isolating phospholipase enzyme, eg phospholipase, sequence variants employing the nucleic acids and polypeptides of the invention. In one aspect, the invention provides variants of a phospholipase gene of the invention, which can be altered by any means including, for example, stochastic or random methods, or, "directed evolution" or non-stochastic, methods, as described above. Isolated variants may be naturally occurring. The variants can also be created in vitro. Variants can be created employing genetic engineering techniques such as site-directed mutagenesis, random chemical mutagenesis, Exonuclease III deletion procedures, and standard cloning techniques. Alternatively, such variants, fragments, analogs or derivatives may be created employing chemical modification or synthesis procedures. Other methods of preparing variants are similarly familiar to those skilled in the art. These include procedures in which nucleic acid sequences obtained from natural isolates are modified to generate nucleic acids encoding polypeptides that have characteristics that increase their value in laboratory and industrial applications. In such procedures, a large number of variant sequences that have one or more nucleotide differences with respect to the sequence obtained from the natural isolate are generated and characterized. These nucleotide differences can result in amino acid changes with respect to the polypeptides encoded by the nucleic acids from the natural isolate. 20 For example, variants can be created using error-prone PCR. In error-prone PCR, PCR is performed under conditions where the copy fidelity of the DNA polymerase is low, such that a high rate of point mutations is obtained over the entire length of the PCR product. Error-prone PCR is described, for example, in 25 Leung, D.W., and the like, Technique, 1:11-15, 1989) and Caldwell, R.C. & Joyce G.F., PCR Methods Applic., 2:28-33, 1992 Briefly, in such procedures, the nucleic acids to be mutagenized are mixed with PCR primers, reaction buffer, MgCh, MnCl 2, Taq polymerase and an appropriate concentration of dNTPs to obtain a high point mutation rate over the entire length of the PCR product. For example, the reaction can be carried out using 20 fmols of nucleic acid to be mutagenized, 30 pmoles of each initiator can be carried out. of PCR, a reaction buffer comprising 50mM KCl, 10mM Tris HCl (pH 8.3) and 0.01% gelatin, 7mM MgCl 2 , 0.5mM MnCI 2 , 5 units Taq polymerase, 0.2mM dGTP, 0.2mM dATP, 1mM dCTP and 1mM dTTP. PCR can be performed for 30 cycles of 94°C for 1 minute, 45°C for 1 minute and 72°C for 1 minute. However, it will be appreciated that these parameters can be varied as appropriate. The mutagenized nucleic acids are cloned into an appropriate vector and the activities of the polypeptides encoded by the mutagenized nucleic acids are evaluated. Variants can also be created using oligonucleotide-directed mutagenesis to generate site-specific mutations in any cloned DNA of interest. Oligonucleotide mutagenesis is described, for example, in Reidhaar-Olson (1988) Science 241:53-57. Briefly, in such procedures a plurality of double-stranded oligonucleotides that undergo one or more mutations to be introduced into the cloned DNA is synthesized and inserted into the cloned DNA to be mutagenized. Clones containing the mutated DNA are recovered and the activities of the polypeptides they encode are evaluated. Another method for generating variants is cluster PCR. Clustering PCR involves assembling a PCR product from a mixture of small DNA fragments. A large number of different PCR reactions take place in parallel in the same vial, with the products of one reaction preparing the products of another reaction. Cluster PCR is described in, for example, U.S. Patent. At the. 5,965,408. Yet another method of generating variants is sexual PCR mutagenesis. In sexual PCR mutagenesis, forced homologous recombination occurs between DNA molecules of different but highly related DNA sequence in vitro as a result of random fragmentation of the DNA molecule based on sequence homology, followed by cross-fixation by primer extension in a PCR reaction. Sexual PCR mutagenesis is described, for example, in Stemmer (1994) Proc. natl. THE- cad. Sci. U.S.A 91:10747 - 10751 . Briefly, in such procedures a plurality of nucleic acids to be recombined are digested with DNase to generate fragments having an average size of 50 - 200 nucleotides. Fragments of the desired medium size are purified and resuspended in a PCR mixture. PCR is conducted under conditions that facilitate recombination between nucleic acid fragments. For example, PCR can be performed by resuspending the purified fragments at a concentration of 10 - 30ng / pl in a solution of 0.2mM of each dNTP, 2.2mM of MgCl 2 , 50mM KCL, 10mM Tris HCl, pH 9.0 and 0.1% Triton X -100. 2.5 Units of Taq polymerase per 100:1 reaction mixture are added and PCR is performed using the following regimen: 94°C for 60 seconds, 94°C for 30 seconds, 50 - 55°C for 30 seconds, 72°C for 30 seconds (30 - 45 times) and 72°C for 5 minutes. However, it will be appreciated that these parameters can be varied as appropriate. In some aspects, oligonucleotides can be included in PCR reactions. In other respects, Kle-now fragments of DNA polymerase I can be used in a first group of PCR reactions and Taq polymerase can be used in a subsequent group of PCR reactions. Recombinant sequences are isolated and the activities of the polypeptides they encode are evaluated. Variants can also be created by mutagen in vivo. In some embodiments, random mutations in a sequence of interest are generated by propagating the sequence of interest in a bacterial strain, such as an E. coli strain, that carries mutations in one or more DNA repair pathways. Such "mutant" strains have a higher random mutation rate than that of a wild-type origin. The propagation of DNA in one of these strains will eventually generate random mutations within the DNA. Mutant strains suitable for use for in vivo mutagenesis are described, for example, in PCT Publication No. WO 91 / 16427. Variants can also be generated using cassette mutagenesis. In cassette mutagenesis, a small region A double-stranded DNA molecule is replaced with a synthetic oligonucleotide "cassette" that differs from the native sequence. The oligonucleotide often contains completely and / or partially randomized native sequence. Recursive ensemble mutagenesis can likewise be employed to generate variants. Recursive ensemble mutagenesis is an algorithm for protein construction (protein mutagenesis) designed to produce diverse populations of phenotypically related mutants whose members differ in amino acid sequence. This method employs a feedback mechanism to control successive rounds of combinatorial cassette mutagenesis. Recursive ensemble mutagenesis is described in, for example, Arkin (1992) Proc. natl. academy Sci. U.S.A. 89:7811-7815. In some embodiments, variants are created using exponential ensemble mutagenesis. Exponential ensemble mutagenesis is a process to generate combinatorial libraries with a high percentage of functional and unique mutants, in which small groups of residues are randomized in parallel to identify, at each altered position, amino acids that lead to functional proteins. Exponential ensemble mutagenesis is described, for example, in Delegrave (1993) Biotechnology Res. 11:1548-1552. Site-directed and random mutagenesis are described, for example, in Arnold (1993) Current Opinion in Biotechnology 4:450-455. In some embodiments, variants are created employing scrambling procedures in which portions of a plurality of nucleic acids encoding distinct polypeptides are fused together to create chimeric nucleic acid sequences encoding chimeric polypeptides as described in, for example, U.S. Patents united n oS 5,965,408, 5,939,250. The invention also provides variants of polypeptides of the invention that comprise sequences in which one or more of the amino acid residues (e.g., from an exemplary polypeptide of the invention) are substituted with a conserved or non-conserved amino acid residue (e.g., a conserved amino acid residue) and such substituted amino acid residue may or may not be the one encoded by the genetic code. Conservative substitutions are those that replace a particular amino acid in a polypeptide with another amino acid with similar characteristics. Thus, polypeptides of the invention include those with conservative substitutions of sequences of the invention, including, but not limited to, the following substitutions: substitutions of an aliphatic amino acid such as Alanine, Valine, Leucine and Isoleucine with another aliphatic amino acid; replacement of a Serine with a Threonine or vice versa; replacing an acidic residue such as Aspartic acid and Glutamic acid with another acidic residue; replacing a residue bearing an amide group, such as Asparagine and Glutamine, with another residue bearing an amide group; exchanging a basic residue such as Lysine and Ar-ginine with another basic residue; and replacing an aromatic residue such as Phenylalanine, Tyrosine with another aromatic residue. Other variants are those in which one or more of the amino acid residues of the polypeptides of the invention include a substituent group. Other variants within the scope of the invention are those in which the polypeptide is associated with another compound, such as a compound to increase the half-life of the polypeptide, for example, polyethylene glycol. Additional variants within the scope of the invention are those in which additional amino acids are fused to the polypeptide, such as a leader sequence, a segregating sequence, a proprotein sequence, or a sequence that facilitates purification, enrichment, or stabilization of the polypeptide. In some aspects, variants, fragments, derivatives and analogs of the polypeptides of the invention retain the same biological function or activity as the exemplary polypeptides, for example, a phospholipase activity, as described herein. In other aspects, the variant, fragment, derivative, or analog includes a proprotein, such that the variant, fragment, derivative or analogue can be activated by cleaving the proprotein moiety to produce an active polypeptide. Codon optimization to obtain high levels of protein expression in host cells. The invention provides methods for modifying phospholipase-encoding nucleic acids to modify codon usage. In one aspect, the invention provides methods for modifying codons in a nucleic acid encoding a phospholipase to increase or decrease its expression in a host cell. The invention likewise provides nucleic acids encoding a phospholipase modified to increase its expression in a host cell, phospholipase enzymes so modified, and methods of making the modified phospholipase enzymes. The method comprises identifying a "non-preferred" or "less preferred" codon in nucleic acid encoding phospholipase and replacing one or more of these non-preferred or less preferred codons with a "preferred codon" encoding the same amino acid as the substituted codon and at least a non-preferred or less preferred codon on the nucleic acid was replaced with a preferred codon encoding the same amino acid. A preferred codon is an over-represented codon encoding sequences in genes in the host cell, and a non-preferred or less preferred codon is one under-represented encoding sequences in genes in the host cell. Host cells for expressing the nucleic acids, expression cassettes and vectors of the invention include bacteria, yeast, fungi, plant cells, insect cells and mammalian cells. Thus, the invention provides methods for optimizing codon usage in all these cells, codon-altered nucleic acids and polypeptides made by codon-altered nucleic acids. Exemplary host cells include gram negative bacteria such as Escherichia coir, gram positive bacteria such as any Baccillus (e.g. 8. cereus) or Streptomyces, Lactobacillus gasseri, Lactococcus lactis, Lactococcus cremoris, Baccillus subtilis. Exemplary host cells likewise include eukaryotic organisms, for example various yeasts such as Saccharomyces sp., including Saccharomyces cerevisiae, Schizosaccharomyces pombe, Pi-chia pastoris, and Kluyveromyces lactis, Hansenula polymorpha, Aspergillus niger, and mammalian cells and cell strains and insect cells and cell strains. Thus, the invention likewise includes nucleic acids and polypeptides optimized for expression in these organisms and species. For example, the codons of a nucleic acid encoding a phospholipase isolated from a bacterial cell are modified such that the nucleic acid is optimally expressed in a bacterial cell other than the bacteria from which the phospholipase was derived, a yeast, a fungus, a cell of plant, an insect cell or a mammalian cell. Methods for optimizing codons are well known in the art, see, for example, US Patent No. 5,795,737; Baca (2000) Int. J. Parasitol. 30:113-118; Hale (1998) Protein Expr. Purify 12:185-188; Narum (2001) Infect. Immun. 69:7250-7253. See similarly Narum (2001) Infect. Im-mun. 69:7250-7253 , describing optimization codons in mouse systems; Outchkourov (2002) Protein Expr. Purify 24:18-24, describing optimization codons in yeast; Feng (2000) Biochemistry 39:15399-15409 , describing optimization codons in E. coli; Humphreys (2000) Protein Expr. Purify 20:252-264, describing use of optimization codon that affects segregation in E. coli. non-human transgenic animals The invention provides transgenic non-human animals comprising a nucleic acid, a polypeptide, an expression vector or cassette, or a transfected or transformed cell of the invention. Transgenic non-human animals can be, for example, goats, rabbits, sheep, pigs, cows, rats and mice, comprising the nucleic acids of the invention. These animals can be used, for example, as in vivo models to study phospholipase activity, or as models to evaluate phospholipase activity modulators in vivo. Coding sequences for the polypeptides to be expressed in transgenic nonhuman animals can be designed to be constitutive, or, under the control of inducible or specific developmental transcriptional factors, vents, fabric specific. Non-human transgenic animals can be designed and generated using any method known in the art; see, for example, United States Patent No. oS 6,211,428, 6,187,992, 6,156,952, 6,118,044, 6,111,166, 6,107,541, 5,959,171, 5,922,854, 5,892,070, 5,880,327, 5,891,698, 5,946. 933, 5,387,742, 5,087,571, describing the preparation and use of transformed cells and transgenic eggs and mice, rats, rabbits, sheep, pigs and cows. See, likewise, for example, Pollock (1999) J. Immunol. Methods 231:147-157, describing the production of recombinant proteins in the milk of transgenic dairy animals; Baguisi (1999) Nat. Biotechnol. 17: 456-461, demonstrating the production of transgenic goats. U.S. Patent At the. 6,211,428, describes the preparation and use of transgenic non-human mammals that express in their brains a nucleic acid construct comprising a DNA sequence. U.S. Patent At the. 5,387,742, describes injecting cloned synthetic and recombinant DNA sequences into fertilized mouse eggs, implanting the injected eggs into pseudopregnant females, and developing to call transgenic mice whose cells express proteins related to the pathology of Alzheimer's disease. United States Patent No. 6,187,992, describes the preparation and use of a transgenic mouse whose genome comprises a disruption of the gene encoding amyloid precursor protein (APP). "Knockout animals" may likewise be employed to practice the methods of the invention. For example, in one aspect, the transgenic or modified animals of the invention comprise a "knockout animal", e.g., a "knockout mouse", not constructed to express or incapable of expressing a phospholipase. Transgenic Plants and Seeds The invention provides transgenic plants and seeds comprising a nucleic acid, a polypeptide (e.g., a phospholipase), an expression vector or cassette, or a transfected or transformed cell of the invention. The invention likewise provides plant products, for example oils, seeds, leaves, extracts and the like, comprising a nucleic acid and / or a polypeptide (e.g. a phospholipase) of the invention. The transgenic plant can be dicotyledonous (a dicot) or monocotyledonous (a monocot). The invention also provides methods of preparing and using these transgenic plants and seeds. The plant cell or transgenic plant expressing a polypeptide of the invention can be constructed according to any method known in the art. See, for example, United States Patent No. 6,309,872. Nucleic acids and expression constructs of the invention can be introduced into a plant cell by any means. For example, nucleic acids or expression constructs can be introduced into the genome of a desired plant host, or, the nucleic acids or expression constructs can be episomes. The introduction into the genome of a desired plant may be such that the host's phospholipase production is regulated by endogenous transcriptional or translational control elements. The invention likewise provides "knockout plants" where insertion of the gene sequence by, for example, homologous recombination, disrupted endogenous gene expression. Means for generating "knockout" plants are well known in the art, see, for example, S-trepp (1998) Proc Natl. academy Know. USA 95:4368-4373; Miao (1995) Plant J 7:359-365. See discussion of transgenic plants, below. The nucleic acids of the invention can be used to impart desired characteristics to essentially any plant, for example, oilseed containing plants such as rice, soybeans, rapeseed, sunflower seeds, sesame and peanuts. Nucleic acids of the invention can be employed to manipulate a plant's metabolic pathways in order to optimize or alter the host's expression of phospholipase. You can change phospholipase activity in a plant. Alternatively, a phospholipase of the invention may be employed in the production of a transgenic plant to produce a compound not naturally produced by that plant. This can lower production costs or create a new product. In one aspect, the first step in the production of a plant transgenic involves the preparation of an expression construct for expression in a plant cell. These techniques are well known in the art. These may include selecting and cloning a promoter, a coding sequence to facilitate efficient binding of ribosomes to mRNA, and selecting the appropriate gene terminator sequences. An exemplary constitutive promoter is CaMV35S, from the cauliflower mosaic virus, which generally results in a high degree of expression in plants. Other promoters are more specific and respond to cues in the plant's internal or external environment. An exemplary light-inducible promoter is the cab gene promoter, encoding the major chlorophyll a / b binding protein. In one aspect, the nucleic acid is modified to obtain greater expression in a plant cell. For example, a sequence of the invention is likely to have a higher percentage of A-T nucleotide pairs compared to those seen in a plant, some of which prefer G-C nucleotide pairs. Therefore, A-T nucleotides in the coding sequence can be replaced with G-C nucleotides without significantly altering the amino acid sequence to enhance production of the gene product in plant cells. Selectable marker gene can be added to the gene construct in order to identify plant tissues or cells that have successfully integrated the transgene. This may be necessary because getting gene incorporation and expression into plant cells is a rare event, occurring in only a few percent of targeted cells or tissues. Selectable marker genes encode proteins that provide resistance to agents that are normally toxic to plants, such as antibiotics or herbicides. Only plant cells that have integrated the selectable marker gene will survive when grown in a medium containing the appropriate antibiotic or herbicide. As for the other inserted genes, marker genes likewise require terminator and promoter sequences for proper function. In one aspect, the preparation of transgenic plants or seeds nicas comprises incorporating the sequences of the invention and, optionally, marker genes into a target expression construct (e.g., a plasmid), along with the positioning of the promoter and its terminator sequences. This may involve transferring the modified gene into the plant by a suitable method. For example, a construct can be introduced directly into plant cell genomic DNA using techniques such as electroporation and microinjection of plant cell protoplasts, or the constructs can be introduced directly into plant tissue using ballistic methods such as particle bombardment. of DNA. For example, see, for example, Christou (1997) Plant Mol. Biol. 35:197-203; Pawlowski (1996) Mol. Biotechnol. 6:17-30; Klein (1987) Nature 327:70-73; Takumi (1997) Genes Genet. syst. 72:63-69, discussing the use of particle bombardment to introduce transgenes into wheat; Adam (1997) supra, for use of particle bombardment to introduce YACs into plant cells. For example, Rinehart (1997) supra, used particle bombardment to generate transgenic cotton plants. Mechanisms for accelerating the particles are described in U.S. Patent. At the. 5,015,580; and, the commercially available PDS-2000 particle acceleration instrument (Biolistics) from BioRad; see likewise, John, US Patent No. 5,608,148; and Ellis, United States Patent No. 5,681,730, describing particle-mediated transformation of gymnosperms In one aspect, protoplasts can be immobilized and injected with nucleic acids, for example, an expression construct. Although plant regeneration from protoplasts is not easy with cereals, plant regeneration is possible in legumes employing somatic embryogenesis from protoplast-derived callus. Organized tissues can be transformed with naked DNA using the gene gun technique, where the DNA is coated in tungsten microprojectiles, fires 1 / 100th the size of the cells, which carries the DNA intensely in the cells and organelles. The transformed tissue is then induced to regenerate, usually by somatic embryogenesis. This technique has been successful in several cereal species including maize and rice. Nucleic acids, for example expression constructs, can be introduced in the same way into plant cells using recombinant viruses. Plant cells can be transformed using viral vectors, such as, for example, vectors derived from tobacco mosaic virus (Rouwendal (1997) Plant Mol. Biol. 33:989-999), see Porta (1996) "Use of replicons viral for the expression of genes in plants," Mol. Biotechnol. 5:209-221. Alternatively, nucleic acids, for example an expression construct, can be combined with suitable T-DNA flanking regions and introduced into a conventional Agrobacterium tumefaciens host vector. The virulence functions of the Agro-bacterium tumefaciens host will direct the insertion of the construct and adjacent marker into the plant cell DNA when the cell is infected by the bacteria. Transformation techniques mediated by Agrobacterium tumefaciens, including unpairing and use of binary vectors, are well described in the scientific literature. See, for example, Horsch (1984) Science 233:496-498; Fraley (1983) Proc. natl. academy Sci. USA 80:4803 (1983); Gene Transfer to Plants, Potrykus, ed. (Springer-Verlag, Berlin 1995). The DNA in an A. tumefaciens cell is contained in the bacterial chromosome as well as another structure known as a Ti (tumor-inducing) plasmid. The Ti plasmid contains a stretch of DNA called T-DNA (~20 kb in length) that is transferred to the plant cell in the process of infection and a series of vir (virulence) genes that direct the infection process. A. tumefaciens can infect only one plant through wounds: when a plant root or stem is injured, it emits certain chemical signals, with respect to which, the genes coming from A. tumefaciens become activated and direct a series of events. necessary for the transfer of the T-DNA from the Ti plasmid to the plant chromosome. T-DNA enters the plant cell through the wound. One speculation is that the T-DNA waits until the plant DNA is being replicated or transcribed, then inserts itself into the exposed plant DNA. In order to employ A. tumefaciens as a transgene vector, the induction section of T-DNA tumor has to be removed, keeping the T-DNA border regions and the genes to come. The transgene is then inserted between the border regions of the T-DNA, where it is transferred into the plant cell and integrated into the plant's chromosomes. The invention provides for the transformation of monocotyledonous plants employing the nucleic acids of the invention, including important cereals, see Hiei (1997) Plant Mol. Biol. 35:205-218. See, likewise, for example, Horsch, Science (1984) 233:496; Fraley (1983) Proc. natl. A-cad. I know USA. 80:4803; Thykjaer (1997) supra; Park (1996) Plant Mol. Biol. 32:1135-1148, discussing integration of T-DNA into genomic DNA. See likewise, D'Halluin, United States Patent No. 5,712,135, describing a process for the stable integration of a DNA comprising a gene that is functional in a cell of a cereal, or other monocotyledonous plant. In one aspect, the third step may involve the selection and regeneration of whole plants capable of transmitting the incorporated target gene to the next generation. Such regeneration techniques rely on the manipulation of certain phytohormones in a tissue culture growth medium, typically relying on a herbicide and / or biocide marker that has been introduced along with the desired nucleotide sequences. Plant regeneration of cultured protoplasts is described in Evans et al., Protoplasts Isolation and Culture, Handbook of Plant Cell Culture, pp. 124-176, MacMillilan Publishing Company, New York, 1983; and Binding, Regeneration of Plants, Plant Protoplasts, pp. 21-73, CRG Press, Boca Raton, 1985. Regeneration can likewise be obtained from plant callus, explants, organs or parts thereof. Such regeneration techniques are generally described in Klee (1987) Ann. Rev. of Plant Phys. 38:467-486. To obtain whole plants from transgenic tissues such as immature embryos, they can be grown under controlled environmental conditions in a series of media containing nutrients and hormones, a process known as tissue culture. As soon as the whole plants are generated and produce seed, the progeny evaluation begins. Once the expression cassette is stably incorporated into transgenic plants, they can be introduced into other plants by sexual crossing. Any of several standard breeding techniques can be employed, depending on the species to be crossed. Since transgenic expression of the nucleic acids of the invention leads to phenotypic changes, plants comprising the recombinant nucleic acids of the invention can be sexually crossed with a second plant to obtain an end product. Thus, the seed of the invention can be derived from a cross between two transgenic plants of the invention, or a cross between a plant of the invention and another plant. Desired effects (e.g., expression of the polypeptides of the invention to produce a plant in which flowering behavior is altered) can be enhanced when both parent plants express the polypeptide (e.g., a phospholipase) of the invention. The desired effects can be passed on to future plant generations by standard pro...

Claims

CLAIMS 1. Ácido nucléico recombinante ou isolado compreendendo uma seqüência de ácido nucléico tendo pelo menos 50% de identidade de se-qüência para SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ IDNO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ IDNO: 111, SEQ ID NO: 113,SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, over a region of at least about 100 residues, in which the nucleic acid encodes at least one polypeptide having phospholipase activity, and the sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection.

2. Recombinant or isolated nucleic acid according to claim 1, wherein the sequence identity is at least about 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63% or 64%.

3. Recombinant or isolated nucleic acid according to the re- Claim 1, wherein the sequence identity is at least about 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, or is 100% sequence identity for SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO.67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ IDNO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ IDNO: 103, SEQ ID NO: 105, SEQ IDNO: 107, SEQ ID NO: 109, SEQ ID NOrlll, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ IDNO: 125, SEQ IDNO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ IDNO: 167, SEQ ID NO: 169, SEQ ID NO: 171 ou SEQ ID NO:

173.

4. Recombinant or isolated nucleic acid according to claim 1, wherein the sequence identity is over a region of at least about 10, 20, 30, 40, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150 or more residues, or the natural size of a gene or transcript.

5. Recombinant or isolated nucleic acid according to king vindicação 1, em que a seqüência de ácido nucléico compreende uma se-qüência como mencionado em SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEO ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119,SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ IDNO: 127, SEQ ID NO: 129, SEQ ID-NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 ou SEQ ID NO: 173., 6. Ácido nucléico recombinante ou isolado de acordo com a reivindicação 1, em que a seqüência de ácido nucléico codifica um polipeptídeo tendo uma seqüência como mencionado em SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ IDNO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ IDNO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO:

174.

7. Recombinant or isolated nucleic acid according to claim 1, wherein the sequence comparison algorithm is a BLAST version 2.2.2 algorithm where a filtering apparatus is set to biastail -p blastp -d "nr pataa" -FF, and all other options are set to default.

8. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises catalyzing hydrolysis of a glycerophosphate ester bond.

9. Recombinant or isolated nucleic acid according to claim 8, wherein the phospholipase activity comprises catalyzing hydrolysis of an ester bond in a phospholipid in a vegetable oil.

10. Recombinant or isolated nucleic acid according to claim 8, wherein the vegetable oil phospholipid comprises an oilseed phospholipid.

11. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises phospholipase C (PLC) activity.

12. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises an activity of phospholipase A (PLA).

13. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises phospholipase B (PLB) activity.

14. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises phospholipase D (PLD) activity.

15. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase D activity comprises either phospholipase D1 or phospholipase D2 activity.

16. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises hydrolysis of a glycoprotein.

17. Recombinant or isolated nucleic acid according to claim 16, wherein the glycoprotein comprises a potato tuber.

18. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises a patata enzymatic activity.

19. Recombinant or isolated nucleic acid according to claim 18, wherein the phospholipase activity comprises a lipid acyl hydrolase (LAH) activity.

20. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity is thermostable.

21. Recombinant or isolated nucleic acid according to claim 20, wherein the polypeptide retains phospholipase activity under conditions comprising a temperature range between about 37°C to about 95°C, or between about 55°C to about 85°C, or between about 70°C to about 75°C, or between about 70°C to about 95°C, or between about 90°C to about 95°C.

22. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity is thermotolerant.

23. Recombinant or isolated nucleic acid according to the re- Claim 22, in which the polypeptide retains phospholipase activity after exposure to a temperature in the range of greater than 37°C to about 95°C, greater than 55°C to about 85°C, or between about 70°C to about 75°C, or greater than 90°C to about 95°C.

24. Ácido nucléico recombinante ou isolado, em que o ácido nu-cléico compreende uma seqüência que hibridiza-se sob condições rigorosas para um ácido nucléico compreendendo SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ IDNO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109,SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, wherein the nucleic acid encodes a polypeptide having phospholipase activity.

25. Recombinant or isolated nucleic acid according to claim 24, wherein the nucleic acid is at least about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 100, 125, 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or more residues in length or the natural size. of the gene or transcript.

26. Recombinant or isolated nucleic acid according to claim 24, wherein the stringent conditions include a washing step comprising washing in 0.2X SSC at a temperature of about 65°C for about 15 minutes.

27. Sonda de ácido nucléico para identificar um ácido nucléico codificando um polipeptídeo com uma atividade de fosfolipase, em que a sonda compreende pelo menos 10 bases consecutivas de uma sequência compreendendo SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ IDNO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103,SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, where the probe identifies the nucleic acid by binding or hybridization.

28. Nucleic acid probe according to claim 27, wherein the probe comprises an oligonucleotide comprising at least about 10 to 50, about 20 to 60, about 30 to 70, about 40 to 80, approximately 60 to 100, or approximately 50 to 150 consecutive bases.

29. Nucleic acid probe for identifying a nucleic acid encoding a polypeptide having phospholipase activity, wherein the probe comprises a nucleic acid comprising at least about 10 consecutive residues of SEQ ID NO: 1; SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ IDNO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 ou SEQ ID NO: 173, em que as identidades de seqüência são determinadas por análise com um algoritmo de comparação de seqüência ou por inspeção visual.

30. Nucleic acid probe according to claim 29, wherein the probe comprises an oligonucleotide comprising at least about 10 to 50, about 20 to 60, about 30 to 70, about 40 to 80, about 60 to 100, or about 50 to 150 consecutive bases.

31. Pair of amplification primer sequences to amplify a nucleic acid encoding a polypeptide having phospho- activity. lipase, wherein the primer pair is capable of amplifying a nucleic acid comprising a sequence as defined in claim 1 or claim 24, or a subsequence thereof.

32. Amplification primer pair according to claim 29, wherein one member of the amplification primer sequence pair comprises an oligonucleotide comprising at least about 10 to 50 consecutive bases, or about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 consecutive bases of the sequence.

33. Par de iniciador de amplificação, em que o par de iniciador compreende um primeiro membro tendo uma seqüência como mencionado em torno da primeira (a 5') 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 ou mais resíduos de SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ IDNO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ IDNO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, and a second member having a sequence such as mentioned around the first (5') 12,13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 or more residues of the complementary filament of the first member.

34. Nucleic acid encoding phospholipase generated by amplification of a polynucleotide using an amplification primer pair as defined in claim 33.

35. Nucleic acid encoding phospholipase according to claim 34, wherein the amplification is by polymerase chain reaction (PCR).

36. Nucleic acid encoding phospholipase according to claim 34, wherein the nucleic acid is generated by amplification of a gene library.

37. Nucleic acid encoding phospholipase according to claim 34, wherein the gene library is an environmental library.

38. Recombinant or isolated phospholipase encoded by a nucleic acid encoding phospholipase as defined in claim 34.

39. Method of amplifying a nucleic acid encoding a polypeptide having phospholipase activity comprising amplification of a standard nucleic acid with an amplification primer sequence pair capable of amplifying a nucleic acid sequence as defined in claim 1 or claim 24, or a subsequence thereof.

40. Method for preparing a phospholipase comprising amplification of a nucleic acid with an amplification primer pair as defined in claim 33 and expression of the amplified nucleic acid.

41. Expression cassette comprising a nucleic acid comprising a sequence as defined in claim 1 or claim 24.

42. Vector comprising a nucleic acid comprising a sequence as defined in claim 1 or claim 24.

43. Cloning vehicle comprising a nucleic acid comprising a sequence as defined in claim 1 or claim 24, wherein the cloning vehicle comprises a viral vector, a plasmid, a phage, a phagomid, a cosmid, a fosmid, a bacteriophage, or an artificial chromosome.

44. Cloning vehicle according to claim 43, wherein the viral vector comprises an adenovirus vector, a viral retrovector or an adeno-associated viral vector.

45. Cloning vehicle according to claim 43, comprising a bacterial artificial chromosome (BAC), a plasmid, a Pi bacteriophage-derived vector (PAC), a yeast artificial chromosome (YAC), or a mammalian artificial chromosome (MAC).

46. ​​Transformed cell comprising a nucleic acid comprising a sequence as defined in claim 1 or claim 24.

47. Transformed cell comprising an expression cassette as defined in claim 41.

48. A cell transformed according to claim 47, wherein the cell is a bacterial cell, a mammalian cell, a fungal cell, a yeast cell, an insect cell, or a plant cell.

49. Transgenic non-human animal comprising a sequence as defined in claim 1 or claim 24.

50. Transgenic non-human animal according to claim 49, wherein the animal is a mouse.

51. Transgenic plant comprising a sequence as defined in claim 1 or claim 24.

52. Transgenic plant according to claim 51, wherein the plant is a maize plant, a sorghum plant, a potato plant, a tomato plant, a wheat plant, an oilseed plant, a rapeseed plant, a soybean plant, a rice plant, a barley plant, a grass, a cottonseed, a palm, a sesame plant, a peanut plant, a sunflower plant or a tobacco plant.

53. Transgenic seed comprising a sequence such as defined in claim 1 or claim 24.

54. Transgenic seed according to claim 53, wherein the seed is a maize seed, a wheat seed, an oilseed, a rapeseed, a soybean seed, a palm seed, a sunflower seed, a sesame seed, a rice seed, a barley seed, a peanut seed, a cotton seed, a palm seed, or a tobacco plant seed.

55. Antisense oligonucleotide comprising a complementary nucleic acid sequence to, or capable of hybridizing under strict conditions to, a sequence as defined in claim 1 or claim 24, or a subsequence thereof.

56. Antisense oligonucleotide according to claim 55, wherein the antisense oligonucleotide is between about 10 to 50, about 20 to 60, about 30 to 70, about 40 to 80, or about 60 to 100 bases in length.

57. Method of inhibiting the translation of a phospholipase message in a cell comprising administering to the cell or expressing in the cell an antisense oligonucleotide comprising a complementary nucleic acid sequence to or capable of hybridizing under strict conditions to a sequence as defined in claim 1 or claim 24.

58. Double-stranded inhibitory RNA (RNAi) molecule comprising a subsequence of a sequence as defined in claim 1 or claim 24.

59. Inhibitory double-stranded RNA (RNAi) molecule according to claim 58, wherein the RNAi is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more double nucleotides in length.

60. A method for inhibiting the expression of a phospholipase in a cell comprising administering to the cell, or expressing in the cell, an inhibitory double-stranded RNA (RNAi), wherein the RNA comprises a subsequence of a sequence as defined in claim 1 or claim 24.

61. Polipeptídeo recombinante ou isolado (i) tendo pelo menos 50% de identidade de seqüência para SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ IDNO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ IDNO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ IDNO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ IDNO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120,SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO: 174, over a region of at least about 100 residues, wherein sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection, or, (ii) encoded by a nucleic acid having at least 50% sequence identity to a sequence as defined in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33,SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: , 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEO ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO;137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, over a region of at least about 100 residues, and the sequence identities are determined by analysis with a sequence comparison algorithm or by visual inspection, or encoded by a nucleic acid capable of hybridizing under strict conditions to a sequence as defined. in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO;53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID ; NO: 129, SEQ ID NO: 131, SEO ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 ou SEQ ID NO:

173.

62. Recombinant or isolated polypeptide according to the re Claim 61, wherein the sequence identity lies over a region of at least about 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72% 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85% 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99% or more, or it is 100% sequence identity.

63. Recombinant or isolated polypeptide according to claim 61, wherein the sequence identity is over a region of at least about 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050 or more residues, or the natural size of an enzyme.

64. Polipeptídeo recombinante ou isolado de acordo com a reivindicação 61, em que o polipeptídeo tem uma seqüência como definido em SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEO ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ IDNO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, or SEQ ID NO:

174.

65. Recombinant or isolated polypeptide according to claim 61, wherein the polypeptide has phospholipase activity.

66. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises catalyzing hydrolysis of a glycerophosphate ester bond.

67. Recombinant or isolated polypeptide according to claim 66, wherein the phospholipase activity comprises catalyzing hydrolysis of an ester bond in a phospholipid in a vegetable oil.

68. Recombinant or isolated polypeptide according to claim 67, wherein the vegetable oil phospholipid comprises an oilseed phospholipid.

69. Recombinant or isolated polypeptide according to claim 67, wherein the vegetable oil phospholipid is derived from a plant oil, a high-phosphorus oil, soybean oil, canola oil, palm oil, cottonseed oil, corn oil, a palm kernel-derived phospholipid, rice bran oil, coconut oil, peanut oil, sesame oil, fish oil, algae phospholipid, sunflower oil, an essential oil, fruit seed oil, grape seed phospholipid, apricot phospholipid, or borage phospholipid.

70. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a phospholipase C (PLC) activity.

71. Recombinant or isolated polypeptide according to the king Claim 65, wherein the phospholipase activity comprises phospholipase A (PLA) activity.

72. Recombinant or isolated polypeptide according to claim 65, wherein a phospholipase activity comprises a phospholipase A1 or phospholipase A2 activity.

73. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a phospholipase D (PLD) activity.

74. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase D activity comprises a phospholipase D1 activity or a phospholipase D2 activity.

75. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises hydrolysis of a glycoprotein.

76. Recombinant or isolated polypeptide according to claim 68, wherein the glycoprotein comprises a potato tuber.

77. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a patatin enzymatic activity.

78. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a lipid acyl hydrolase (LAH) activity.

79. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity is thermostable.

80. Recombinant or isolated polypeptide according to claim 79, wherein the polypeptide retains phospholipase activity under conditions comprising a temperature range of from about 20 to about 30°C, from about 25°C to about 40°C, from about 37°C to about 95°C, from about 55°C to about 85°C, from about 70°C to about 95°C, from about 70°C to about 75°C, or from about 90°C to about 95°C.

81. Recombinant or isolated polypeptide according to the king Claim 65, in which the phospholipase activity is thermotolerant.

82. Recombinant or isolated polypeptide according to claim 81, wherein the polypeptide retains phospholipase activity after exposure to a temperature in the range greater than 37°C to about 95°C, greater than 55°C to about 85°C, between about 70°C to about 75°C, or greater than 90°C to about 95°C.

83. Recombinant or isolated polypeptide comprising a polypeptide as defined in claim 61, and not having a signal sequence.

84. Recombinant or isolated polypeptide comprising a polypeptide as defined in claim 61, and having a heterologous signal sequence.

85. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a specific activity at about 37°C in the range of about 10 to about 100 units per milligram, about 100 to about 1000 units per milligram of protein, about 500 to about 750 units per milligram of protein, about 500 to about 1200 units per milligram of protein, or about 750 to about 1000 units per milligram of protein.

86. Recombinant or isolated polypeptide according to claim 81, wherein the thermotolerance comprises retention of at least half of the specific phospholipase activity at 37°C after being heated to an elevated temperature.

87. Recombinant or isolated polypeptide according to claim 81, wherein the thermotolerance comprises retention of specific activity at 37°C in the range of about 500 to about 1200 units per milligram of protein after being heated to an elevated temperature.

88. Recombinant or isolated polypeptide according to claim 61, wherein the polypeptide comprises at least one glycosylation site.

89. Recombinant or isolated polypeptide according to claim 88, wherein the glycosylation is an N-linked glycosylation.

90. Recombinant or isolated polypeptide according to claim 89, wherein the polypeptide is glycosylated after being expressed in a P. pastoris or an S. pombe.

91. Recombinant or isolated polypeptide according to claim 65, wherein the polypeptide retains phospholipase activity under conditions comprising approximately pH 6.5, pH 6.0, pH 5.5, 5.0, pH 4.5 or 4.

0.

92. Recombinant or isolated polypeptide according to claim 65, wherein the polypeptide retains phospholipase activity under conditions comprising approximately pH 7.5, pH 8.0, pH 8.5, pH 9, pH 9.5, pH 10 or pH 10.

5.

93. Protein preparation comprising a polypeptide as defined in claim 61, wherein the protein preparation comprises a liquid, a solid or a gel.

94. Heterodimer comprising a polypeptide as defined in claim 61 and a second domain.

95. Heterodimer according to claim 94, wherein the second domain is a polypeptide and the heterodimer is a fusion protein.

96. Heterodimer according to claim 94, wherein the second domain is an epitope or a label.

97. Homodimer comprising a polypeptide as defined in claim 61.

98. Immobilized polypeptide, wherein the polypeptide comprises a sequence as defined in claim 61, or a subsequence thereof.

99. Immobilized polypeptide according to claim 98, wherein the polypeptide is immobilized on a cell, a metal, a resin, a polymer, a ceramic, a glass, a microelectrode, a graphite particle, a bead, a gel, a plate, an array or a capillary tube.

100. Arrangement comprising an immobilized polypeptide as defined in claim 61.

101. An arrangement comprising an immobilized nucleic acid as defined in claim 1 or claim 24.

102. Recombinant or isolated antibody that specifically binds to a polypeptide as defined in claim 61.

103. Recombinant or isolated antibody according to claim 102, wherein the antibody is a monoclonal or polyclonal antibody.

104. Hybridoma comprising an antibody that specifically binds to a polypeptide as defined in claim 61.

105. Method of isolating or identifying a polypeptide with phospholipase activity comprising the steps of: (a) provide an antibody as defined in claim 102; (b) provide a sample comprising polypeptides; and (c) contact the sample from step (b) with the antibody from step (a) under conditions in which the antibody can specifically bind to the polypeptide, thereby isolating or identifying a polypeptide having phospholipase activity.

106. Method of preparing an anti-phospholipase antibody comprising administering to a non-human animal a nucleic acid as defined in claim 1 or claim 24, or a subsequence thereof, in an amount sufficient to generate a humoral immune response, thereby forming an anti-phospholipase antibody.

107. Method of preparing an anti-phospholipase antibody comprising administering to a non-human animal a polypeptide as defined in claim 61, or a subsequence thereof, in an amount sufficient to generate a humoral immune response, thereby forming an anti-phospholipase antibody.

108. Method of producing a recombinant polypeptide comprising the steps of: (a) provide a nucleic acid operably linked to a promoter, wherein the nucleic acid comprises a sequence as defined in claim 1 or claim 24; and (b) express the nucleic acid from step (a) under conditions that allow the expression of the polypeptide, thereby producing a recombinant polypeptide.

109. Method according to claim 108, also comprising transforming a host cell with the nucleic acid of step (a) followed by expression of the nucleic acid of step (a), thereby producing a recombinant polypeptide in the transformed cell.

110. Method for identifying a polypeptide that has phospholipase activity comprising the following steps: (a) providing a polypeptide as defined in claim 65; (b) providing a phospholipase substrate; and (c) contact of the polypeptide with the substrate of step (b) and detection of a decrease in the amount of substrate or an increase in the amount of a reaction product, wherein a decrease in the amount of substrate or an increase in the amount of reaction product detects a polypeptide that has phospholipase activity.

111. Method for identifying a phospholipase substrate comprising the following steps: (a) providing a polypeptide as defined in claim 65; (b) provision of a test substrate; and (c) contact of the polypeptide from step (a) with the test substrate from step (b) and detection of a decrease in the amount of substrate or an increase in the amount of reaction product, wherein a decrease in the amount of substrate or an increase in the amount of a reaction product identifies the test substrate as a phospholipase substrate.

112. Method for determining whether a test compound binds specifically to a polypeptide comprising the following steps: (a) express a nucleic acid or a vector comprising nucleic acid under conditions permissive for nucleic acid translation into a polypeptide, wherein the nucleic acid has a sequence such as- defined in claim 1 or claim 24; (b) provide a test compound; (c) contact the polypeptide with the test compound; and (d) determine whether the test compound from step (b) binds specifically to the polypeptide.

113. Method for determining whether a test compound binds specifically to a polypeptide, comprising the following steps: (a) provide a polypeptide as defined in claim 61; (b) provide a test compound; (c) contact the polypeptide with the test compound; and (d) determine whether the test compound from step (b) binds specifically to the polypeptide.

114. Method for identifying a modulator of phospholipase activity comprising the following steps: (a) providing a polypeptide as defined in claim 65; (b) provision of a test compound; (c) contact of a polypeptide from step (a) with the test compound from step (b), and evaluation of phospholipase activity, wherein the change in phospholipase activity measured in the presence of the test compound compared to the activity in the absence of the test compound provides a determination that the test compound modulates phospholipase activity.

115. A method according to claim 114, wherein phospholipase activity is evaluated by providing a phospholipase substrate and detecting a decrease in the amount of substrate or an increase in the amount of a reaction product, or an increase in the amount of substrate or a decrease in the amount of a reaction product.

116. Method according to claim 115, wherein a decrease in the amount of substrate or an increase in the amount of reaction product with the test compound when compared to the amount of The substrate or reaction product without the test compound identifies the test compound as an activator of phospholipase activity.

117. A method according to claim 115, wherein an increase in the amount of substrate or a decrease in the amount of reaction product with a test compound when compared with the amount of substrate or reaction product without the test compound identifies the test compound as an inhibitor of phospholipase activity.

118. Computer system comprising a processor and a data storage device, wherein said data storage device has stored on it a polypeptide sequence or a nucleic acid sequence, wherein the polypeptide sequence comprises a sequence as defined in claim 61, a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

119. Computer system according to claim 118, also comprising a sequence comparison algorithm and a data storage device having at least one reference sequence stored therein.

120. Computer system according to claim 119, wherein the sequence comparison algorithm comprises a computer program that indicates polymorphisms.

121. Computer system according to claim 119, also comprising an identifier that identifies one or more features in said sequence.

122. A computer-interpretable medium having stored therein a polypeptide sequence or a nucleic acid sequence, wherein the polypeptide sequence comprises a polypeptide as defined in claim 61; a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

123. Method for identifying a feature in a sequence comprising the step of: (a) reading the sequence using a computer program that identifies one or more features in a sequence, wherein the sequence comprises a polypeptide sequence or a nucleic acid sequence, wherein the polypeptide sequence comprises a polypeptide as defined in claim 61; a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; and (b) identify one or more features in the sequence with the computer system.

124. Method for comparing a first sequence with a second sequence comprising the steps of: (a) reading the first sequence and the second sequence using a computer program that compares the sequences, wherein the first sequence comprises a polypeptide sequence or a nucleic acid sequence, wherein the polypeptide sequence comprises a polypeptide as defined in claim 61 or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; and (b) determining the differences between the first sequence and the second sequence using the computer program.

125. Method according to claim 124, wherein the step of determining the differences between the first sequence and the second sequence also comprises the step of identifying polymorphisms.

126. Method according to claim 124, also comprising an identifier that identifies one or more features in a sequence.

127. Method according to claim 126, comprising reading the first sequence using a computer program and identifying one or more features in the sequence.

128. Method for isolating or recovering a nucleic acid encoding a polypeptide with phospholipase activity from an environmental sample comprising the steps of: (a) providing a pair of amplification initiator sequences as defined in claim 33; (b) isolation of a nucleic acid from the environmental sample or treatment of the environmental sample, so that the nucleic acid in the sample be accessible for hybridization to the amplification initiator pair; and, (c) combining the nucleic acid from step (b) with the amplification primer pair from step (a) and amplifying the nucleic acid from the environmental sample, thereby isolating or recovering a nucleic acid encoding a polypeptide with phospholipase activity from an environmental sample.

129. Método de acordo com a reivindicação 128, em que cada membro do par de seqüência de iniciador de amplificação compreende um oligonucleotídeo que compreende pelo menos cerca de 10 a 50 bases consecutivas de uma seqüência como definido em SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ IDNO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ IDNO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ IDNO: 97, SEQ ID NO: 99, SEQ ID NO: 101,SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, or a subsequence of these., 130. Method for isolating or recovering a nucleic acid encoding a polypeptide with phospholipase activity from a sample environmental impact assessment encompassing the following stages: (a) providing a polynucleotide probe comprising a sequence as defined in claim 1 or claim 24, or a subsequence thereof; (b) isolation of a nucleic acid from the environmental sample or treatment of the environmental sample such that the nucleic acid in the sample is accessible for hybridization to a step (a) polynucleotide probe; (c) combination of the isolated nucleic acid or treated environmental sample from step (b) with the polynucleotide probe from step (a); and (d) isolation of a nucleic acid that specifically hybridizes with the polynucleotide probe from step (a), thereby isolating or recovering a nucleic acid encoding a polypeptide with phospholipase activity from an environmental sample.

131. Method according to claim 128 or claim 130, wherein the environmental sample comprises a water sample, a liquid sample, a soil sample, an air sample or a biological sample.

132. Method according to claim 131, wherein the biological sample is derived from a bacterial cell, a protozoan cell, an insect cell, a yeast cell, a plant cell, a fungal cell or a mammalian cell.

133. A method for generating a nucleic acid variant encoding a polypeptide with phospholipase activity comprising the following steps: (a) providing a standard nucleic acid comprising a sequence as defined in claim 1 or claim 24; and (b) modifying, deleting or adding one or more nucleotides to the standard sequence, or a combination thereof, to generate a variant of the standard nucleic acid.

134. Method according to claim 133, also comprising variant nucleic acid expression to generate a variant phospholipase polypeptide.

135. A method according to claim 133, wherein the modifications, additions or deletions are introduced by a method comprising error-prone PCR, shuffling, oligonucleotide-directed mutagenesis, assembly PCR, sex PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive assembly mutagenesis, exponential assembly mutagenesis, site-specific mutagenesis, gene reassembly, Gene Site Saturation Mutagenesis® (GSSM®), synthetic linkage reassembly (SLR) and a combination thereof.

136. A method according to claim 133, wherein the modifications, additions or deletions are introduced by a method comprising recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, uracil-containing pattern mutagenesis, closed duplex mutagenesis, point linkage imperfection repair mutagenesis, repair-deficient host strain mutagenesis, chemical mutagenesis, radiogenic mutagenesis, deletion mutagenesis, restriction selection mutagenesis, restriction purification mutagenesis, artificial gene synthesis, assembly mutagenesis, creation of chimeric nucleic acid multimers and a combination thereof.

137. Method according to claim 133, wherein the method is iteratively repeated until a phospholipase having an altered or different activity or an altered or different stability from that of a polypeptide encoded by a standard nucleic acid is produced.

138. Method according to claim 137, wherein the variant phospholipase polypeptide is thermotolerant, and retains some activity after being exposed to an elevated temperature.

139. Method according to claim 137, wherein the variant phospholipase polypeptide has increased glycosylation when compared to phospholipase encoded by a standard nucleic acid.

140. Method according to claim 137, wherein the variant phospholipase polypeptide has phospholipase activity under elevated temperature, whereas the standard nucleic acid-encoded phospholipase is inactive under elevated temperature.

141. Method according to claim 133, wherein the method is iteratively repeated until a phospholipase coding sequence having an altered codon usage from that of the standard nucleic acid is produced.

142. Method according to claim 133, wherein the method is iteratively repeated until a phospholipase gene having a higher or lower level of message expression or stability than that of the standard nucleic acid is produced.

143. A method for modifying codons in a nucleic acid encoding a polypeptide with phospholipase activity to increase its expression in a host cell, the method comprising the following steps: (a) provide a nucleic acid encoding a polypeptide with phospholipase activity comprising the sequence as defined in claim 1 or claim 24; and, (b) identify a non-preferred or less preferred codon in the nucleic acid from step (a) and replacing it with a preferred or neutrally employed codon encoding the same amino acid as the substituted codon, wherein a preferred codon is a codon that is over-represented in coding sequences in genes in the host cell and a non-preferred or less preferred codon is a codon that is under-represented in coding sequences in genes in the host cell, thereby modifying the nucleic acid to increase its expression in a host cell.

144. Method for modifying codons in a nucleic acid encoding a phospholipase polypeptide, the method comprising the following steps: (a) providing a nucleic acid encoding a polypeptide with phospholipase activity comprising a sequence as defined in claim 1 or claim 24; and, (b) identifying a codon in the nucleic acid from step (a) and replacing it with a different codon encoding the same amino acid as the replaced codon, thereby modifying the codons in an acid nucleic acid that codes for a phospholipase.

145. A method for modifying codons in a nucleic acid encoding a phospholipase polypeptide to increase its expression in a host cell, the method comprising the following steps: (a) providing a nucleic acid encoding a phospholipase polypeptide comprising a sequence as defined in claim 1 or claim 24; and, (b) identification of a non-preferred or less preferred codon in the nucleic acid from step (a) and replacement of it with a preferred or neutrally employed codon encoding the same amino acid as the replaced codon, where a preferred codon is a codon that is over-represented in coding sequences in genes in the host cell and a non-preferred or less preferred codon is a codon that is under-represented in coding sequences in genes in the host cell, thereby modifying the nucleic acid to increase its expression in a host cell.

146. A method for modifying a codon in a nucleic acid encoding a polypeptide to decrease its expression in a host cell, the method comprising the following steps: (a) providing a nucleic acid encoding a phospholipase polypeptide comprising a sequence as defined in claim 1 or claim 24; and, (b) identification of at least one preferred codon in the nucleic acid from step (a) and replacement of it with a non-preferred or less preferred codon encoding the same amino acid as the replaced codon, where a preferred codon is a codon that is over-represented in coding sequences in genes in a host cell and a non-preferred or less preferred codon is a codon that is under-represented in coding sequences in genes in the host cell, thereby modifying the nucleic acid to increase its expression in a host cell.

147. Method according to claim 146, wherein the host cell is a bacterial cell, a fungal cell, an insect cell, a yeast cell, a plant cell or a mammalian cell.

148. Method for producing a nucleic acid library encoding a plurality of modified phospholipase active sites or substrate binding sites, wherein the modified active sites or substrate binding sites are derived from a first nucleic acid comprising a sequence encoding a first active site or a first substrate binding site, the method comprising the following steps: (a) provision of a first nucleic acid encoding a first active site or first substrate binding site, wherein the first nucleic acid sequence comprises a sequence that hybridizes under stringent conditions to a sequence as defined in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 1I, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 97,SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 127, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO-.145, SEQ IDNO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161, SEQ ID NO: 163, SEQ ID NO: 165, SEQ ID NO: 167, SEQ ID NO: 169, SEQ ID NO: 171 or SEQ ID NO: 173, or a subsequence thereof, and the nucleic acid encodes a phospholipase active site or a binding site of , phospholipase substrate; (b) providing a group of mutagenic oligonucleotides that encode naturally occurring amino acid variants in a plurality of labeled codons in the first nucleic acid and, (c) use of the mutagenic oligonucleotide group to generate a variant nucleic acid group encoding substrate binding site or active site encoding, encoding a range of amino acid variations in each amino acid codon that has been mutagenized, thereby producing a nucleic acid library encoding a plurality of modified phospholipase active sites or substrate binding sites.

149. Method according to claim 148, comprising mutagenizing the first nucleic acid of step (a) by a method comprising an optimized directed evolution system, Gene Site Saturation Mutagenesis® (GSSM®), or a synthetic linkage reassembly (SLR).

150. Method according to claim 148, comprising mutagenizing the first step nucleic acid (a) or variants by a method comprising error-prone PCR, shuffling, oligonucleotide-directed mutagenesis, assembly PCR, sex PCR mutagenesis, in vivo mutagenesis, cassette mutagenesis, recursive assembly mutagenesis, exponential assembly mutagenesis, site-specific mutagenesis, gene reassembly, Gene Site Saturation Mutagenesis® (GSSM®), synthetic linkage reassembly (SLR) and a combination thereof.

151. Method according to claim 148, comprising mutagenizing the first step nucleic acid (a) or variants by a method comprising recombination, recursive sequence recombination, phosphothioate-modified DNA mutagenesis, uracil-containing pattern mutagenesis, closed duplex mutagenesis, point linkage imperfection repair mutagenesis, repair-deficient host strain mutagenesis, restriction selection mutagenesis, restriction purification mutagenesis, artificial gene synthesis, ensemble mutagenesis, creation of A chimeric nucleic acid multimer and a combination thereof.

152. Method for preparing a small molecule comprising the following steps: (a) providing a plurality of biosynthetic enzymes capable of synthesizing or modifying a small molecule, wherein one of the enzymes comprises a phospholipase enzyme encoded by a nucleic acid comprising a sequence as defined in claim 1 or claim 24; (b) provision of a substrate during at least one of the enzyme steps (a); and (c) reaction of the substrate from step (b) with the enzymes under conditions that facilitate a plurality of biocatalytic reactions to generate a small molecule by a series of biocatalytic reactions.

153. Method for modifying a small molecule, comprising the following steps: (a) providing a phospholipase enzyme, wherein the enzyme comprises a polypeptide as defined in claim 65, or a nucleic acid-encoded polypeptide comprising a nucleic acid sequence as defined in claim 1 or claim 24; (b) supplying a small molecule; and (c) reaction of the enzyme from step (a) with a small molecule from step (b) under conditions that facilitate an enzymatic reaction catalyzed by the phospholipase enzyme, thereby modifying a small molecule by a phospholipase enzymatic reaction.

154. Method according to claim 153, comprising a plurality of small molecule substrates for the enzyme in step (a), thereby generating a library of modified small molecules produced by at least one enzymatic reaction catalyzed by the phospholipase enzyme.

155. Method according to claim 153, also comprising a plurality of additional enzymes under conditions that facilitate a plurality of biocatalytic reactions by the enzymes to form a library of small, modified molecules produced by a plurality of enzymatic reactions.

156. Method according to claim 155, also comprising the step of testing the library to determine whether a particular modified small molecule exhibiting a desired activity is present in the library.

157. Method according to claim 156, wherein the library testing step also comprises the steps of systematically eliminating all but one of the biocatalytic reactions employed to produce a portion of the plurality of modified small molecules within the library to test the modified small molecule portion for the presence or absence of the particular modified small molecule with a desired activity, and identifying at least one specific biocatalytic reaction that produces the particular modified small molecule of desired activity.

158. Method for determining a functional fragment of a phospholipase enzyme comprising the following steps: (a) providing a phospholipase enzyme, wherein the enzyme comprises a polypeptide as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; and (b) deletion of a plurality of amino acid residues from the sequence in step (a) and testing of the remaining subsequence for phospholipase activity, thereby determining a functional fragment of a phospholipase enzyme.

159. Method according to claim 158, wherein phospholipase activity is evaluated by providing a phospholipase substrate and detecting a decrease in the amount of substrate or an increase in the amount of a reaction product.

160. Method for constructing whole-cell novel or modified phenotypes using real-time metabolic flux analysis, the method comprising the following steps: (a) preparation of a modified cell by modifying the genetic composition of a cell, wherein the genetic composition is modified by adding to the cell a nucleic acid comprising a sequence as defined in claim 1 or claim 24; (b) culture of the modified cell to generate a plurality of modified cells; (c) evaluation of at least one metabolic parameter of the cell by monitoring the cell culture from step (b) in real time; and, (d) analysis of step (c) data to determine whether the measured parameter differs from a comparable assessment in an unmodified cell under similar conditions, thereby identifying a phenotype constructed in the cells using real-time metabolic flux analysis.

161. A method according to claim 160, wherein the genetic makeup of the cell is modified by a method comprising deleting a sequence or modifying a sequence in the cell, or reducing the expression of a gene.

162. Method according to claim 160, also comprising selecting a cell comprising a newly constructed phenotype.

163. Method according to claim 162, also comprising cultivating the selected cell, thereby generating a new cell strain comprising a newly constructed phenotype.

164. Seqüências sinal isolada ou recombinante consistindo em uma seqüência como definido nos resíduos 1 a 16, 1 to 17, 1 a 18, 1 a 19, 1 a 20, 1 a 21, 1 a 22, 1 a 23, 1 a 24, 1 a 25, 1 a 26, 1 a 27, 1 a 28, 1 a 28, 1 a 30 ou 1 a 31, 1 a 32 ou 1 a 33 de SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEO ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108 SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 124, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138; SEQ ID NO: 140; SEQ ID NO: 142; SEQ ID NO: 144; NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 170, SEQ ID NO: 172, ou SEQ ID NO:

174.

165. Chimeric polypeptide comprising at least one first domain comprising a signal peptide (SP) having a sequence as defined in claim 164, and at least one second domain comprising a polypeptide or heterologous peptide, wherein the polypeptide or heterologous peptide is not naturally associated with the signal peptide (SP).

166. Chimeric polypeptide according to claim 165, wherein the heterologous polypeptide or peptide is not a phospholipase.

167. Chimeric polypeptide according to claim 165, wherein the heterologous polypeptide or peptide is amino-terminal α, carboxy-terminal α or both ends of the signal peptide (SP) or a catalytic domain (CD).

168. Recombinant or isolated nucleic acid encoding a chimeric polypeptide, wherein the chimeric polypeptide comprises at least one first domain comprising a signal peptide (SP) having a sequence as defined in claim 164 and at least one second domain comprising a heterologous polypeptide or peptide, wherein the heterologous polypeptide or peptide is not naturally associated with the signal peptide (SP).

169. Method of increasing the thermotolerance or thermostability of a phospholipase polypeptide, the method comprising glycosylating a phospholipase, wherein the polypeptide comprises at least thirty contiguous amino acids of a polypeptide as defined in claim 61, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24, thereby increasing the thermotolerance or thermostability of the phospholipase.

170. A method of overexpressing a recombinant phospholipase in a cell comprising expressing a vector comprising a nucleic acid sequence as defined in claim 1 or claim 24, wherein the overexpression is accomplished by use of a high-activity promoter, a dicistronic vector, or by vector gene amplification.

171. Method of preparing a transgenic plant comprising the following steps: (a) introduction of a heterologous nucleic acid sequence into the cell, wherein the heterologous nucleic acid sequence comprises a sequence as defined in claim 1 or claim 24, thereby producing a transformed plant cell; (b) production of transgenic plant from transformed cell.

172. Method as defined in claim 171, wherein the step (a) also includes introducing the heterologous nucleic acid sequence by electroporation or microinjection of plant cell protoplasts.

173. Method as defined in claim 171, wherein the step (a) involves introducing the heterologous nucleic acid sequence directly into plant tissue by DNA particle bombardment or by using an Agrobacterium tumefaciens host.

174. Method of expressing a heterologous nucleic acid sequence in a plant cell comprising the following steps: (a) transform the plant cell with a heterologous nucleic acid sequence operably linked to a promoter, where the sequence A heterologous nucleic acid comprises a sequence as defined in claim 1 or claim 24; (b) cultivate the plant under conditions in which the heterologous nucleic acid sequence is expressed in the plant cell.

175. A method for hydrolyzing, breaking down or disrupting a composition comprising phospholipid, comprising the following steps: (a) providing a polypeptide having a phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) providing a composition comprising a phospholipid; and (c) contact the polypeptide of step (a) with the e-step (b) composition under conditions in which the phospholipase hydrolyzes, breaks down or disrupts the composition comprising phospholipid.

176. Method as defined in claim 175, wherein the composition comprises a lipid bilayer or membrane comprising phospholipid.

177. Method as defined in claim 175, wherein the composition comprises a plant cell, a bacterial cell, a yeast cell, an insect cell, or an animal cell.

178. Method for liquefying or removing a composition comprising phospholipid comprising the following steps: (a) providing a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) providing a composition comprising a phospholipid; and (c) contact of the polypeptide from step (a) with the composition from step (b) under conditions in which the phospholipase removes or liquefies the composition comprising phospholipid.

179. Detergent composition comprising a polypeptide as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24, wherein the polypeptide has phospholipase activity.

180. Detergent composition according to claim 179, wherein the phospholipase is either a non-surface-active phospholipase or a surfactant phospholipase.

181. Detergent composition according to claim 179, wherein the phospholipase is formulated in a non-aqueous liquid composition, a melt solid, a lyophilized powder, a granular form, a particulate form, a pressed tablet, a pellet, a gel form, a paste, an aerosol, or a suspension form.

182. Method for washing an object comprising the following steps: (a) supplying a composition comprising a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; b) providing an object; and (c) contact of the polypeptide from step (a) and the object from step (b) under conditions in which the composition may wash away the object.

183. A method for degumming an oil comprising the following steps: (a) supplying a composition comprising a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) supplying a composition comprising a fat or oil containing phospholipid; and (c) contact of the polypeptide from step (a) and the step composition (b) under conditions in which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition.

184. Method according to claim 183, wherein the oil-comprising composition comprises a plant oil or fat, animal, algae or fish.

185. Method according to claim 184, wherein the plant oil comprises rice bran oil, soybean oil, rapeseed oil, corn oil, palm kernel oil, canola oil, sunflower oil, sesame oil or peanut oil.

186. Method according to claim 183, wherein the polypeptide hydrolyzes a phosphatide from a hydratable and / or non-hydratable phospholipid in the composition comprising oil.

187. Method according to claim 183, wherein the polypeptide hydrolyzes a phosphatide at a glyceryl phosphoester linkage to generate a water-soluble phosphate compound and diglyceride.

188. Method according to claim 183, wherein the polypeptide has phospholipase C activity.

189. Method according to claim 183, wherein the polypeptide has phospholipase D activity and a phosphatase enzyme is also added.

190. Method according to claim 183, wherein the contact comprises hydrolysis of a hydrated phospholipid in an oil.

191. Method according to claim 183, wherein the hydrolysis conditions of step (c) comprise a temperature of about 20°C to 40°C at an alkaline pH.

192. Method according to claim 190, wherein the alkaline conditions comprise a pH of about pH 8 to pH 10.

193. Method according to claim 183, wherein the hydrolysis conditions of step (c) comprise a reaction time of about 3 to 10 minutes.

194. Method according to claim 183, wherein the hydrolysis conditions of step (c) comprise hydrolysis of hydratable and non-hydratable phospholipids in oil at a temperature of about 50°C to 60°C, at a pH of about pH 5 to pH 6.5, at a pH of about pH 6.0 to pH 7.5, or at a pH of about pH 5 to pH 8.0, employing a time reaction time of approximately 30 to 60 minutes.

195. Method according to claim 183, wherein the polypeptide is attached to a filter and the phospholipid-containing fat or oil is passed through the filter.

196. Method according to claim 183, wherein the polypeptide is added to a solution comprising phospholipid-containing fat or oil and then the solution is passed through a filter.

197. Method for converting a non-hydratable phospholipid into a hydratable form comprising the following steps: (a) providing a composition comprising a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) providing a composition comprising a non-hydratable phospholipid; and (c) contact of the polypeptide from step (a) and the composition of step (b) under conditions in which the polypeptide converts the non-hydratable phospholipid into a hydratable form.

198. Method according to claim 197, wherein the polypeptide has phospholipase C activity.

199. Method according to claim 197, wherein the polypeptide has phospholipase D activity and a phosphatase enzyme is similarly added.

200. Method for caustic refining of a phospholipid-containing composition comprising the following steps: (a) providing a composition comprising a polypeptide having phospholipase activity; (b) providing a composition comprising a phospholipid; and (c) contact of the polypeptide from step (a) with the composition from step (b) before, during or after caustic refinement.

201. Method according to claim 200, wherein the poly- The peptide has phospholipase C activity.

202. Method according to claim 200, wherein the polypeptide having phospholipase activity is added before the addition of acid or caustic.

203. Method according to claim 200, wherein the polypeptide having phospholipase activity is added during caustic refining and varying levels of acid and caustic are added depending on the phosphorus levels and free fatty acid levels.

204. Method according to claim 200, wherein the polypeptide having phospholipase activity is added after caustic refining: in an intense mixer or holding mixer before separation; following a heating step; in a centrifuge; in a soap feedstock; in a wash water; or during the bleaching and deodorizing steps.

205. Method for purifying a phytosterol or a triterpene comprising the following steps: (a) providing a composition comprising a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) providing a composition comprising a phytosterol or a triterpene; and (c) contact of the polypeptide from step (a) with the composition from step (b) under conditions in which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition.

206. Method according to claim 205, wherein the polypeptide has phospholipase C activity.

207. Method according to claim 205, wherein the phytosterol or a triterpene comprises a plant sterol.

208. Method according to claim 207, wherein the plant sterol is derived from a vegetable oil.

209. Method according to claim 208, wherein the oil Vegetable oil includes coconut oil, canola oil, cocoa butter oil, corn oil, cottonseed oil, linseed oil, olive oil, palm oil, peanut oil, oil derived from rice bran, safflower oil, sesame oil, soybean oil, or sunflower oil.

210. Method according to claim 205, comprising using non-polar solvents to quantitatively extract free phytosterols and phytosterol fatty acid esters.

211. Method according to claim 205, wherein the phytosterol or a triterpene comprises a p-sitosterol, a campesterol, an es-tigmasterol, an estigmastanol, p-sitostanol, a sitostanol, a desmosterol, a calinasterol, a poriferasterol, a clionasterol or a brassicasterol.

212. Method for refining crude oil comprising the following steps: (a) providing a composition comprising a polypeptide having phospholipase activity as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) supplying a composition comprising an oil comprising a phospholipid; and (c) contact of the polypeptide from step (a) with the composition from step (b) under conditions in which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition.

213. Method according to claim 212, wherein the polypeptide has phospholipase C activity.

214. Method according to claim 212, wherein the polypeptide having phospholipase activity is in an aqueous solution that is added to the composition.

215. Method according to claim 214, wherein the water level is between about 0.5 to 5%.

216. Method according to claim 214, wherein the processing time is less than about 2 hours.

217. Method according to claim 216, wherein the processing time is less than about 60 minutes.

218. Method according to claim 217, wherein the processing time is less than about 30 minutes, less than about 15 minutes, or less than about 5 minutes.

219. Method according to claim 212, wherein the hydrolysis conditions comprise a temperature between about 25°C-70°C.

220. Method according to claim 212, wherein the hydrolysis conditions comprise the use of caustics.

221. Method according to claim 212, wherein the hydrolysis conditions comprise a pH between about pH 3 and pH 10.

222. Method according to claim 212, wherein the hydrolysis conditions comprise the addition of emulsifiers and / or mixing after contact of step (c).

223. Method according to claim 212, comprising adding an emulsion stopper and / or heating or cooling to promote separation of an aqueous phase.

224. Method according to claim 212, comprising degumming before the contact step to collect lecithin by centrifugation and then adding a PLC, a PLC and / or a PLA to remove non-hydratable phospholipids.

225. Method according to claim 212, comprising degumming crude oil in water to less than 10 ppm phosphorus for edible oils and subsequent physical refining to less than about 50 ppm phosphorus for biodiesel oils.

226. Method according to claim 212, comprising adding acid to promote hydration of non-hydratable phospholipids.

227. Method for removing a fat or grease comprising the following steps: (a) supply of a composition comprising a poly- a peptide having phospholipase activity as mentioned in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24, wherein the phospholipase activity comprises phospholipase D activity and a phosphatase enzyme; (b) supplying a composition comprising an oil or fat containing phospholipid; and (c) contact of the polypeptide from step (a) and the compound from step (b) under conditions in which the polypeptide can catalyze the hydrolysis of a phospholipid in the compound.

228. Composition having the equivalent of a phospholipase C activity comprising providing a composition comprising a polypeptide having a phospholipase activity as defined according to claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24, wherein the phospholipase activity comprises a phospholipase D activity, and a phosphatase enzyme.

229. Method for improving or preventing lipopolysaccharide (LPS)-mediated toxicity comprising administering to a patient a pharmaceutical composition comprising a polypeptide as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

230. Method for detoxifying an endotoxin comprising contacting the endotoxin with a polypeptide as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

231. Method for deacylating a 2' or 3' fatty acid chain from a lipid A comprising contacting lipid A with a polypeptide as defined in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

232. Method according to claim 231, wherein the poly- The peptide has patatin activity.

233. Method according to claim 232, wherein the patatin has a sequence as mentioned in SEQ ID NO: 12 (encoded by SEQ ID NO:11), SEQ ID NO:14 (encoded by SEQ ID NO:13), SEQ ID NO:18 (encoded by SEQ ID NO:17), SEQ ID NO:26 (encoded by SEQ ID NO:25), SEQ ID NO:28 (encoded by SEQ ID NO:27), SEQ ID NO:34 (encoded by SEQ ID NO:33), SEQ ID NO:36 (encoded by SEQ ID NO:35), SEQ ID NO:44 (encoded by SEQ ID NO:43), SEQ ID NO:46 (encoded by SEQ ID NO:45), SEQ ID NO:56 (encoded by SEQ ID NO:55), SEQ ID NO:60 (encoded by SEQ ID NO:59), SEQ ID NO:66 (encoded by SEQ ID NO:65), SEQ ID NO:72 (encoded by SEQ ID NO:71), SEQ ID NO:78 (encoded by SEQ ID NO:77), SEQ ID NO:87 (encoded by SEQ ID NO:86), SEQ ID NO:88 (encoded by SEQ ID NO:87), SEQ ID NO:92 (encoded by SEQ ID NO:91), SEQ ID NO:96 (encoded by SEQ ID NO:95), SEQ ID NQ:100 (encoded by SEQ ID NO:99), SEQ ID NQ:104 (encoded by SEQ ID NO:103),SEQ ID NO:126 (encoded by SEQ ID NO:125), SEQ ID NO:128 (encoded by SEQ ID NO:127), SEQ ID NO:132 (encoded by SEQ ID NO:131), SEQ ID NO:134 (encoded by SEQ ID NO:133), SEQ ID NO:136 (encoded by SEQ ID NO:135), or SEQ ID NO:138 (encoded by SEQ ID NO:137).

234. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises a combination of one or more phospholipase activities.

235. Recombinant or isolated nucleic acid according to claim 234, wherein the phospholipase activity comprises PLC and PLA activity; PLB and PLA activity; PLC and PLD activity; PLC and PLB activity; PLB and patatin activity; PLC and patatin activity; PLD and PLA; PLD, PLA, PLB and PLC activity; or PLD, PLA, PLB, PLC and patatin activity.

236. Recombinant or isolated nucleic acid according to claim 234, wherein the phospholipase activity comprises lysophospholipase (LPL) activity or lysophospholipase-transacylase (LPTA) activity. or lysophospholipase (LPL) activity and lysophospholipase-transacylase (LPTA) activity.

237. Recombinant or isolated nucleic acid according to claim 1, wherein the phospholipase activity comprises catalyzing the hydrolysis of a glycerol phosphate ester linkage into phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and / or phosphatidic acid or a combination thereof.

238. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises a combination of one or more phospholipase activities.

239. Recombinant or isolated polypeptide according to claim 238, wherein the phospholipase activity comprises PLC and PLA activity; PLB and PLA activity; PLC and PLD activity; PLC and PLB activity; PLB and patatin activity; PLC and patatin-PLD and PLA activity; PLD, PLA, PLB and PLC activity; or PLD, PLA, PLB, PLC and patatin activity.

240. Recombinant or isolated polypeptide according to claim 238, wherein the phospholipase activity comprises lysophospholipase (LPL) activity or lysophospholipase-transacylase (LPTA) activity or lysophospholipase (LPL) activity and lysophospholipase-transacylase (LPTA) activity.

241. Recombinant or isolated polypeptide according to claim 65, wherein the phospholipase activity comprises catalyzing the hydrolysis of a glycerol phosphate ester linkage into phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and / or phosphatidic acid or a combination thereof.

242. Method according to claim 183, wherein the polypeptide having a phospholipase activity has a PLC activity and the method enhances neutral oils.

243. Method according to claim 183, wherein the polypeptide having phospholipase activity has PLC activity and the method increases diacylglycerol (DAG) production to contribute to a oil phase.

244. Method according to claim 183, also comprising adding one or more polypeptides having a protease, an amylase, a lipase, a cutinase, another phospholipase, a carbohydrase, a cellulase, a pectinase, a mannanase, an arabinase, a galactanase, a xylanase, an oxidase, for example, a lactase, and / or a peroxidase or polypeptides with equivalent activity, or a combination thereof, to also destroy gum mass and enhance oil production.

245. Method according to claim 183, wherein the hydrolysis conditions comprise an alkaline pH.

246. Method according to claim 183, wherein alkaline conditions are sufficient to cause isomerization of a 1,2-DAG produced by a PLC into a 1,3-DAG.

247. Method according to claim 183, also comprising the physical removal of gum produced by the degumming method by the addition of a hardening substance.

248. Method according to claim 247, wherein the hardening substance comprises talc.

249. Method according to claim 183, wherein the final degummed oil product is enriched in 1,3-DAG.

250. Method according to claim 249, wherein the final degummed oil product comprises not less than 1.0% of 1,3-DAG.

251. Process for reducing gum mass and increasing neutral oil (triglyceride) gain through reduced oil capture comprising the following steps: (a) provide a composition comprising a polypeptide having phospholipase activity as mentioned in claim 65, or a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24; (b) provide a composition comprising an oil or fat containing phospholipid; and (c) contact the polypeptide from step (a) and the composition of step (b) under conditions in which the polypeptide can catalyze the hydrolysis of a phospholipid in the composition for a time sufficient to reduce the gum mass and increase neutral oils.

252. Protein preparation according to claim 93, wherein the protein preparation comprises a formulation comprising a non-aqueous liquid composition, a molten solid, a powder, a lyophilized powder, a granular form, a particulate form, a pressed tablet, a pellet, a pill, a gel form, a hydrogel, a paste, an aerosol, a spray, a lotion, a suspension formulation, an aqueous / oily emulsion, a cream, a capsule, a vesicle or a micellar suspension.

253. Method according to claim 175, comprising use of high shear mixing of the composition, followed by no or low shear mixing with at least one polypeptide of the invention having phospholipase activity to allow adequate contact of the phospholipid substrate with the phospholipase.

254. Method according to claim 200, wherein caustic refining conditions are generated by adding a concentrated caustic solution.

255. Method according to claim 254, wherein the concentrated caustic solution is more concentrated than the industrial standard of 11%.

256. Method according to claim 255, wherein the concentrated caustic solution is between about 12% and 50% concentrated.

257. Method according to claim 200, wherein the polypeptide having phospholipase activity has a sequence as mentioned in claim 65, or comprises a polypeptide encoded by a nucleic acid as defined in claim 1 or claim 24.

258. Method according to claim 200, wherein the composition comprising the phospholipid comprises a plant.

259. Method according to claim 200, wherein the polypeptide is transgenically expressed in the plant.

260. Method according to claim 200, wherein the polypeptide having phospholipase activity is added during the crushing of a seed or other plant part, or the polypeptide having phospholipase activity is added following crushing or before refining.

261. Method according to claim 200, comprising a process as defined in Figure 13.

262. Method according to claim 226, wherein sufficient acid is added to promote the reduction of the magnesium and calcium metal content.

263. Method for preparing a variant phospholipase coding sequence that has increased expression in a host cell comprising modifying a sequence as mentioned according to claim 1 or claim 22 such that one, several or all N-linked glycosylation site coding motifs are modified into a non-glycosylated motif.

264. Isolated, synthetic or recombinant phospholipase encoded by a sequence made by the method as defined in claim 263.

265. Method for preparing a variant phospholipase coding sequence encoding a phospholipase that has increased resistance to a protease comprising modifying an amino acid equivalent to position 131 of SEQ ID NO: 2 in one, several or all of the following residues: Lysine (K); Serine (S); Glycine (G); Arginine (R); Glutamine (Q); Alanine (A); Isoleucine (I); Histidine (H); Phenylalanine (F); Threonine (T); Methionine (M); Leucine (L).

266. Isolated, synthetic or recombinant phospholipase encoded by a sequence made by the method as defined in claim 265.

267. Method for preparing a variant phospholipase coding sequence encoding a phospholipase that has decreased resistance to a protease comprising modifying an amino acid equivalent to the posi- tion 131 of SEQ ID NO: 2 in one, several or all of the following residues: Tryptophan (W); Glutamate (E); Tyrosine (Y).

268. Isolated, synthetic or recombinant phospholipase encoded by a sequence made by the method as defined in claim 267.

269. Method for preparing and expressing a protein that has a biological activity whose activity is temporarily inactivated by glycosylation comprising: (a) providing a nucleic acid that encodes a protein that has biological activity, wherein the protein is not naturally glycosylated; (b) insertion of at least one coding sequence for a glycosylation motif into the protein-coding nucleic acid, wherein the glycosylated form of the protein is inactive; (c) insertion of a targeting sequence into the protein such that it is directed to the host cell's segregation track, where the host cell is able to recognize the glycosylation motif and glycosylate the protein; and (d) expression of the modified nucleic acid in the host cell.

270. Method according to claim 269, also comprising deglycosylating expressed protein, thereby reactivating protein activity.

271. Method according to claim 270, wherein the host cell is a eukaryotic cell.

272. Method for expressing phospholipase C comprising (a) provide a Pichia strain with a Mut phenotype + ; (b) insert nucleic acid encoding heterologous phospholipase C into the Pichia- strain, and, (c) cultivate the Pichia strain under conditions in which phospholipase C is expressed.

273. Method according to claim 272, also comprising supplementing the growing conditions with zinc.

274. Cell system for expressing phospholipase C comprising dendo a strain of Pichia with a Mut phenotype +comprising a heterologous phospholipase C-encoding nucleic acid operably linked to an operable promoter in the Pichia strain.

275. Cell system for expressing a heterologous protein 5 comprising a zeocin-resistant Pichia strain cell.

276. Zeocin-resistant yeast cell system for expressing a heterologous protein comprising the steps of (a) provide a Pichia sp cell comprising a heterologous nucleic acid capable of expressing a heterologous protein; 10 (b) cultivate the cell under conditions comprising zeocin in an initial concentration; (c) select cells resistant to the initial zeocin concentration, and reculture them under conditions comprising a higher zeocin concentration; and 15 (d) select the cells grown in step (c) resistant to con higher concentration of zeocin.

277. Zeocin-resistant yeast cell system according to claim 276, wherein the heterologous protein is an enzyme, or optionally, a phospholipase, or optionally a phospholipase C (PLC).