Method for producing perillaldehyde and / or perillyl lactone

Abstract

The present invention discloses a method for producing perillaldehyde and / or perillyl lactone, comprising: contacting a first component with a second component to produce perillaldehyde and / or perillyl lactone; wherein the first component is selected from components containing lysine-2-diol, and the second component is selected from enzymes or microorganisms or their cell lysates or enzyme extracts capable of converting lysine-2-diol into perillaldehyde and / or perillyl lactone.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 202512060524.0, filed on December 31, 2025, entitled "Method for producing perillaldehyde and / or perillaldehyde lactone".

[0002] RELEVANT APPLIANCES This application claims priority to Chinese patent application CN202412000066.7, filed on December 31, 2024, which is incorporated herein by reference in its entirety. Technical Field

[0003] This invention relates to the field of biotechnology, and in particular to methods and compositions for producing perillaldehyde or perillyl lactone by biotransformation. Background Technology

[0004] Sclareolide is a sesquiterpene-like compound, appearing as a white crystalline powder insoluble in water but readily soluble in organic solvents, possessing a distinctive aroma. Due to its unique fragrance, sclareolide is widely used in the flavoring and fragrance industry, especially as a tobacco flavoring agent. Furthermore, it is widely used in the food industry to enhance the sensory properties of food, improving flavor and aroma. Another important use of sclareolide is as a precursor in the synthesis of ambergris—a substitute for ambroxol. Sclareolide glycol, also known as ambroxol, has a similar aroma to ambergris and can be used as a fixative in the formulation of everyday fragrances; it is also an important intermediate in the synthesis of ambroxol.

[0005] The main methods for obtaining sclareol and sclarediol are chemical synthesis. Currently, industrial production mainly uses sclareol as raw material, which is oxidized by an oxidant to generate sclareol, and then reduced to sclarediol. The obtained sclarediol can be dehydrated and cyclized under the action of organic acids, sulfonyl chloride compounds, or Lewis acids to obtain ambroxol (Synthetic Chemistry, 2018, 26(1): 55-65). Although the current industrial production route has undergone multiple optimizations and process adjustments, it still faces some problems that cannot be ignored. First, the waste treatment cost generated during the oxidation and reduction of sclareol is high, which puts considerable pressure on the economic benefits and environmental impact of enterprises. In addition, the supply fluctuations and price instability of sclareol raw materials also bring great uncertainty to production. Therefore, the development of new production methods for sclareol and sclarediol has attracted the attention of researchers.

[0006] In recent years, the biological production of perillaldehyde and perillyl glycol has attracted attention. At the end of the 20th century, Farboood MI et al. discovered ATCC 20624 (… Hyphozyma roseoniger ATCC 20624) can convert styraxol or labdenediol to styraxol (US4798799A); Cryptococcus lightifolius ( Cryptococcus albidus ATCC 20918 can convert perillyl alcohol to perillyl lactone (US4970163A; US5155029A; US5212078A). In the decades that followed, various wild fungi were found to catalyze the conversion of perillyl alcohol to perillyl lactone or ambroxol (CN 113293106 B; CN 113502306 B); and to convert perillyl alcohol to perillyl lactone or perillyl diol (CN101426900B). Besides directly utilizing wild fungi for biotransformation, in recent years researchers have also produced perillyl alcohol through genetically engineered bacteria or perillyl lactone or perillyl diol through mixed culture. In 2012, Michel Schalk et al. discovered... Clary sage Two key enzymes for the synthesis of perillaldehyde, lysine pyrophosphate synthase (SsLPPs3) and perillaldehyde synthase (SsTPs1132), catalyze the reactions of geranyl pyrophosphate (GGPP) to lysine pyrophosphate (LPP) and LPP to perillaldehyde, respectively. A biosynthetic pathway for perillaldehyde was constructed using *E. coli* as the host, and combined with high-density fermentation technology, the yield of perillaldehyde reached 1.5 g / L. J. Am. Chem. Soc. 2012, 134(46):18900-18903). In 2023, Ni He et al. established a co-culture system of strains with different pathway modules, and used engineered Saccharomyces cerevisiae YN42 to produce perillaldehyde, which can then be processed by ATCC 20624 ( Hyphozyma roseoniger ATCC 20624) was used as a carbon source to convert salicylic acid to salicylic acid, with a maximum yield of 644 mg / L. ACS Sustainable Chem. Eng. 2023, 11, 1939 1948). In 2024, Tang DD et al. used engineered brewing yeast ( Saccharomyces cerevisiae ) and Cryptococcus lightensis ( Cryptococcus albidus A modular co-culture system was constructed using ATCC 20918. Glucose was converted to perillyl alcohol using engineered yeast, and then perillyl alcohol was converted to perillyl lactone using Cryptococcus lighti. By optimizing the solubilizer, adjusting the inoculum ratio, and improving the culture temperature, the yield of perillyl lactone reached a maximum of 626.3 mg / L. J. Agric. Food Chem. 2024, 72, 19977 19984).

[0007] Overall, there are few existing studies on the biosynthesis of perillyl glycol or perillyl lactone, and most of them use perillyl alcohol as an intermediate for subsequent biotransformation. More diverse biosynthetic methods need to be explored in depth to provide new ideas and references for the efficient and green synthesis of perillyl glycol, perillyl lactone, and ambroxol. Summary of the Invention

[0008] This invention provides a method for producing perillaldehyde and / or perillyl lactone, comprising: The following components are contacted: component (1) and component (2) are brought together to produce perillaldehyde and / or perillyl lactone; wherein, The first component is selected from components containing lysine diol. The second component is selected from enzymes or microorganisms or their cell lysates or enzyme extracts that can convert lysine diol into perillyl diol and / or perillyl lactone.

[0009] In some embodiments, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from the genera *Hyphozyma* or *Cryptococcus*. In some embodiments, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from... Hyphozyma rose-colored Or Cryptococcus lightensis ( Cryptococcus albidus ).

[0010] In some embodiments, the microorganisms capable of converting lysine-3-diol into perillylene-3-diol and / or perillylene lactone are selected from fungi with accession number ATCC 20624, fungi with accession number ATCC 20918, or derivatives thereof.

[0011] In some embodiments, the component containing lysinediol includes isolated lysinediol, a mixture containing lysinediol, a microorganism capable of producing lysinediol or its culture medium, supernatant, cell lysate, or extract.

[0012] In some embodiments, the component containing lysinediol is a microorganism capable of producing lysinediol or its culture medium, supernatant, cell lysate, or extract.

[0013] In some embodiments, the method is used to produce perillaldehyde, or to produce perillaldehyde diol and perillaldehyde.

[0014] In some embodiments, the method is used to produce perillaldehyde, wherein the microorganism capable of converting lysine-3-diol into perillaldehyde and / or perillaldehyde lactone is selected from fungi or derivatives thereof with accession number ATCC 20918.

[0015] In some embodiments, the microorganism capable of producing lysine diol is a recombinant engineered bacterium capable of producing lysine diol.

[0016] In some embodiments, the recombinant engineered bacteria capable of producing lysine-6-diol contains a phosphatase that catalyzes the production of lysine-6-diol pyrophosphate (LPP) to lysine-6-diol (LOH). This phosphatase is derived from Mg(2+)-dependent phosphatidate phosphatase, phosphatidic acid phosphatase type 2, serine / threonine-protein phosphatase, HAD-like hydrolase superfamily, phosphate (PA) phosphatase, polyphosphoinositide phosphatase, serine / threonine-protein kinases, or alkaline phosphatase. Phosphatase, phosphoprotein phosphatase family, serine / threonine-protein phosphatases, SIT4 phosphatase-associated protein family, TAP42 / TAP46-like superfamily, dual-specificity lipid and protein phosphatase, polynucleotide kinase 3 phosphatase, 5'-deoxynucleotidase, myosin family, histidine phosphatase superfamily, S-2-haloalkanoic acid dehalogenase, phosphatidylglycerol phosphatase Mutase, halogen dehalogenase (HAD) hydrolase (HAD-hydrolase superfamily)The following families are included in the phosphatase family: phosphatase-related phosphoesterase family, squalene / phytoene synthase family, terpene cyclases, phosphatidate phosphatase App1, protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), halogen dehalogenase (HAD)-like sugar phosphate phosphatase, Fig. 4-like polyphosphoinositide phosphatase, casein kinase 1 (Se / Thr protein kinase), lipoprotein family, and PPZ / Ppq1 family. The family includes the clade-1 family, the HAD-IA hydrolase family, the HAD-IF subfamily, the dolichyldiphosphatase family, and the haloperoxidase superfamily. In some embodiments, the phosphorylase is phosphatidylphosphatase (APP1), polyacyl diphosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), inositol polyphosphate phosphatase (FIG4), 1-glycerol phosphate phosphatase 1 (GPP1), casein kinase I (HRR25), phosphatidylphosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), serine / threonine protein phosphatase (PTC3), serine / threonine protein phosphatase (PTC3), SIT4-associated protein SAP155, SIT4-associated protein SAP185, 2A Phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase (TEP1), polynucleotides3'-phosphatase (TPP1), phosphorylhydrolase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphorylhydrolase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), phosphatidylglycerol phosphatase (PgpA), phosphatidylglycerol phosphatase (PgpB), phosphatidylglycerol phosphatase (PgpC), carbapenyl diphosphatase (YbjG), dihydroxy diphosphatase (DOLPP1), phosphorylhydrolase (PLPP6), farnesyl diphosphatase (YisP), farnesol synthase (TPS2), acyclic sesquiterpene synthase (TPS1), or farnesol synthase (TPS13).

[0017] In some embodiments, the phosphatase is a phosphatase derived from a microorganism or a functional variant thereof, the microorganism including bacteria, yeast, or fungi, the bacteria being selected from Escherichia or Bacillus, the yeast being selected from Saccharomyces, or the phosphatase being derived from plants or humans (…). Homo sapiens The phosphorylase is a phosphorylase or a functional variant thereof derived from bacteria, yeast, or fungi, wherein the bacteria are selected from *Escherichia coli* (E. coli). Escherichia coli Bacillus subtilis ( Bacillus subtilis The yeast is brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), or the phosphorylase is derived from plants or humans ( Homo sapiens Phosphorylase or a functional variant thereof, wherein the plant is selected from moso bamboo ( Phyllostachys edible ),corn( Corn ) or japonica rice ( Oryza sativa subsp. japonica In some embodiments, the phosphorylase is derived from *Saccharomyces cerevisiae* (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), Escherichia coli ( Escherichia to be cultivated ), human beings ( Homo sapiens Bacillus subtilis ( Bacillus subtilis ),bamboo( Phyllostachys edible ),corn( Corn ) or japonica rice ( Oryza sativa subsp. japonica Phosphohydrolases or their functional variants.

[0018] In some embodiments, the phosphorylase comprises an amino acid sequence selected from SEQ ID NO. 11-226, or an amino acid sequence having at least 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% sequence identity with an amino acid sequence selected from SEQ ID NO. 11-226. In some embodiments, the phosphorylase comprises a protein selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.2 ... ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID NO.226, or an amino acid sequence selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160. SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID The amino acid sequence of NO. 226 has at least 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% sequence identity.

[0019] In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO.237-452, or by a nucleotide sequence having at least 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% sequence identity with a nucleotide sequence selected from SEQ ID NO.237-452. In some embodiments, the phosphorylase is selected from SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO.271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO. NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ ID NO.452, or the nucleotide sequence selected from SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO.271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385. SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ IDNO. 452 encodes a nucleotide sequence with at least 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% sequence identity.

[0020] In some embodiments, the recombinant engineered bacteria capable of producing lysine diol also contains genes for the lysine diol pyrophosphate (LPP) synthesis pathway.

[0021] In some embodiments, the lysine pyrophosphate (LPP) synthesis pathway gene includes the farnesyl pyrophosphate (FPP) synthesis pathway gene, the geraniol geraniol pyrophosphate synthase (GGPPS) gene, and the lysine pyrophosphate synthase (LPPS) gene.

[0022] In some embodiments, the farnesyl pyrophosphate (FPP) synthesis pathway genes include one or more of the following: acetyl-CoA transferase gene, 3-methyl-3-hydroxyglutaryl-CoA synthase gene, 3-hydroxy-3-methylglutaryl-CoA reductase gene, mevalonate kinase gene, mevalonate-5-phosphate kinase gene, mevalonate-5-pyrophosphate decarboxylase gene, pentene pyrophosphate isomerase gene, or farnesyl pyrophosphate synthase gene.

[0023] In some embodiments, the recombinant engineered bacteria capable of producing lysine diol further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the activity of geraniol geraniol pyrophosphate synthase (GGPPS); and (2) At least one gene modification that enhances the activity of lysine pyrophosphate diol ester synthase (LPPS).

[0024] In some implementation schemes, wherein, At least one gene modification that enhances the activity of gerany-gerany-gerany pyrophosphate synthase (GGPPS) includes: introducing an exogenous gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, increasing the gene copy number of the gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, and / or replacing the natural promoter of the endogenous gerany-gerany pyrophosphate synthase (GGPPS) with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of lysandrindiol pyrophosphate synthase (LPPS) includes: introducing an exogenous lysandrindiol pyrophosphate synthase (LPPS) gene, increasing the gene copy number of the lysandrindiol pyrophosphate synthase (LPPS) gene, and / or replacing the natural promoter of the endogenous lysandrindiol pyrophosphate synthase (LPPS) with a promoter that has a higher expression level.

[0025] In some implementation schemes, wherein, The exogenous gerany-gerany pyrophosphate synthase (GGPPS) comprises the amino acid sequence shown in SEQ ID NO. 9, preferably encoded by the nucleotide sequence of SEQ ID NO. 235; and / or The exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a truncated lysine pyrophosphate diol ester synthase (LPPS); preferably, the truncated lysine pyrophosphate diol ester synthase (LPPS) is a signal peptide with the first 63 amino acids removed; preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) comprises the amino acid sequence shown in SEQ ID NO. 10; more preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) is encoded by the nucleotide sequence of SEQ ID NO. 236.

[0026] In some embodiments, the recombinant engineered bacteria capable of producing lysine diol further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the function of acetyl-CoA transferase; (2) At least one gene modification that enhances the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase; (4) At least one gene modification that enhances the activity of mevalonate kinase; (5) At least one gene modification that enhances the activity of mevalonate-5-phosphate kinase; (6) At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase; (7) At least one gene modification that enhances the activity of pentylene pyrophosphate isomerase; and (8) At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase.

[0027] In some implementation schemes, wherein, At least one gene modification that enhances the function of acetyl-CoA transferase includes: introducing an exogenous acetyl-CoA transferase gene, increasing the gene copy number of the acetyl-CoA transferase gene, and / or replacing the natural promoter of endogenous acetyl-CoA transferase with a promoter that has a higher expression level. At least one gene modification that enhances the function of 3-methyl-3-hydroxyglutaryl-CoA synthase includes: introducing an exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene, increasing the gene copy number of the 3-methyl-3-hydroxyglutaryl-CoA synthase gene, and / or replacing the natural promoter of endogenous 3-methyl-3-hydroxyglutaryl-CoA synthase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase includes: introducing an exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene, increasing the gene copy number of the 3-hydroxy-3-methylglutaryl-CoA reductase gene, and / or replacing the natural promoter of endogenous 3-hydroxy-3-methylglutaryl-CoA reductase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of mevalonate kinase includes: introducing an exogenous mevalonate kinase gene, increasing the gene copy number of the mevalonate kinase gene, and / or replacing the natural promoter of endogenous mevalonate kinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-phosphokinase includes: introducing an exogenous mevalonate-5-phosphokinase gene, increasing the gene copy number of the mevalonate-5-phosphokinase gene, and / or replacing the natural promoter of endogenous mevalonate-5-phosphokinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase includes: introducing an exogenous mevalonate-5-pyrophosphate decarboxylase gene, increasing the gene copy number of the mevalonate-5-pyrophosphate decarboxylase gene, and / or replacing the natural promoter of endogenous mevalonate-5-pyrophosphate decarboxylase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of pentyrene pyrophosphate isomerase includes: introducing an exogenous pentyrene pyrophosphate isomerase gene, increasing the copy number of the pentyrene pyrophosphate isomerase gene, and / or replacing the natural promoter of the endogenous pentyrene pyrophosphate isomerase with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase includes: introducing an exogenous farnesyl pyrophosphate synthase gene, increasing the gene copy number of the farnesyl pyrophosphate synthase gene, and / or replacing the natural promoter of endogenous farnesyl pyrophosphate synthase with a promoter that has a higher expression level.

[0028] In some implementation schemes, wherein: The exogenous acetyl-CoA transferase comprises the amino acid sequence shown in SEQ ID NO.3, preferably, the exogenous acetyl-CoA transferase is encoded by the nucleotide sequence of SEQ ID NO.229; The exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase comprises the amino acid sequence shown in SEQ ID NO.4, preferably, the exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase is encoded by the nucleotide sequence of SEQ ID NO.230; The exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a truncated 3-hydroxy-3-methylglutaryl-CoA reductase, wherein the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is a truncated 3-hydroxy-3-methylglutaryl-CoA reductase with 528 amino acids removed from its N-terminus; preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase comprises the amino acid sequence shown in SEQ ID NO. 2, and preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is encoded by the nucleotide sequence of SEQ ID NO. 228; The exogenous mevalonate kinase comprises the amino acid sequence shown in SEQ ID NO.5, preferably, the exogenous mevalonate kinase is encoded by the nucleotide sequence of SEQ ID NO.231; The exogenous mevalonate-5-phosphokinase comprises the amino acid sequence shown in SEQ ID NO. 6, preferably, the exogenous mevalonate-5-phosphokinase is encoded by the nucleotide sequence of SEQ ID NO. 232; The exogenous mevalonate-5-pyrophosphate decarboxylase comprises the amino acid sequence shown in SEQ ID NO.7, preferably, the exogenous mevalonate-5-pyrophosphate decarboxylase is encoded by the nucleotide sequence of SEQ ID NO.233; The exogenous pentene pyrophosphate isomerase comprises the amino acid sequence shown in SEQ ID NO. 8, preferably, the exogenous pentene pyrophosphate isomerase is encoded by the nucleotide sequence of SEQ ID NO. 234; and / or The exogenous farnesyl pyrophosphate synthase comprises the amino acid sequence shown in SEQ ID NO.1, preferably encoded by the nucleotide sequence of SEQ ID NO.227.

[0029] In some embodiments, the recombinant engineered bacteria capable of producing lysine diol are bacteria or yeast; preferably, the bacteria are selected from Escherichia, Corynebacterium, or Bacillus, and the yeast is selected from Saccharomyces, Pichia, Hansenula, Kluyveromyces, Phaffia, Schizosaccharomyces, Candida, Yarrowia, Hyphozyma, or Cryptococcus; more preferably, the bacteria are selected from Escherichia coli (…). Escherichia coli ), Corynebacterium glutamicum ( Corynebacterium glutamicum ), or Bacillus subtilis ( Bacillus subtilis The yeast is selected from brewer's yeast (Saccharomyces cerevisiae). Saccharomyces yeast Pichia pastoris () Shepherd's pie ), Phaffia colomata ( Komagataellaphaffii) , Saccharomyces cerevisiae ( Schizosaccharomyces pombe ), Candida albicans ( Candida albicans ), Candida utilis ( Useful Candida ), Yarrowia lipolytica ( Yarrow lipolytic ), Hansenula polymorpha ( Hansenula polymorpha Pichia pastoris (Canada) Peach Canadian ), Max Kluyveromycin ( Kluyveromyces marxianus Kluyveromycin (lactic acid yeast) Kluyveromyces lactis ), Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus white ) or Red Pavlova yeast ( Phaffia rhodozyma ).

[0030] In some embodiments, the microorganisms capable of producing lysinediol and the microorganisms capable of converting lysinediol into perillyl and / or perillyl lactone are co-cultured under suitable conditions.

[0031] In some embodiments, the cell density ratio of the microorganisms capable of producing lysinediol and the microorganisms capable of converting lysinediol to perillaldehyde and / or perillaldehyde lactone is in the range of 20:1 to 1:20; preferably, the cell density ratio of the microorganisms capable of producing lysinediol and the microorganisms capable of converting lysinediol to perillaldehyde and / or perillaldehyde lactone is 10.5:1 or 8.5. :1, 4.8:1, 3.9:1, 2:1, 1.9:1, 1.55:1, 1:1, 0.96:1, 0.78:1, 1:2, 1:2.1, 1:2.58, 1:5.2, 1:6.45, 1:11.5, 1:14; More preferably, the microorganisms capable of producing lysoprenediol and the microorganisms capable of converting lysoprenediol into perillyl alcohol and / or perillyl lactone have the accession number ATCC. The cell density ratios of the fungi in the mixture of 20624 are 8.5:1, 3.9:1, 2:1, 1.55:1, 1:1, 0.78:1, 1:2, 1:2.58, and 1:6.45; or the cell density ratios of the fungi in the mixture of the microorganisms capable of producing lysine-1-diol and the microorganisms capable of converting lysine-1-diol into perillyl alcohol and / or perillyl lactone are 10.5:1, 4.8:1, 2:1, 1.9:1, 1:1, 0.96:1, 1:2, 1:2.1, 1:5.2, and 1:11.5.

[0032] In some embodiments, the temperature of the mixed culture is above 15°C; preferably, the temperature of the mixed culture is 15°C-40°C, and more preferably, the temperature of the mixed culture is 25°C-30°C.

[0033] In some embodiments, the method includes adding at least one carbon source during the mixed culture process, said at least one carbon source being selected from glucose, fructose, sucrose, acetic acid, glycerol, maltose, lactic acid, and succinic acid.

[0034] In some embodiments, the microorganisms capable of converting lysinediol to perillyl and / or perillyl lactone are either lysinediol-induced or not lysinediol-induced before being mixed with the microorganisms capable of producing lysinediol.

[0035] In some embodiments, the method further includes separating perillaldehyde and / or perillyl lactone.

[0036] Another aspect of the present invention provides a composition comprising the following two microorganisms: First microorganism: Selected from microorganisms capable of producing lysine diol; Second microorganism: Selected from microorganisms capable of converting lysine-enrichediol into perillyl alcohol and / or perillyl lactone.

[0037] In some embodiments, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from... Hyphozyma rose-colored Or Cryptococcus lightensis ( Cryptococcus albidus ).

[0038] In some embodiments, the microorganisms capable of converting lysine-3-diol into perillylene-3-diol and / or perillylene lactone are selected from fungi with accession number ATCC 20624 or fungi with accession number ATCC 20918 or their derivatives.

[0039] In some embodiments, the first microorganism comprises a phosphatase that catalyzes the formation of lysandrindiol pyrophosphate (LPP) to lysandrindiol (LOH). The phosphatase is derived from Mg(2+)-dependent phosphatidate phosphatase, phosphatidic acid phosphatase type 2, serine / threonine-protein phosphatase, HAD-like hydrolase superfamily, phosphate (PA) phosphatase, polyphosphoinositide phosphatase, serine / threonine-protein kinases, alkaline phosphatase, or phosphaprotein phosphatase. This includes various protein phosphatases, serine / threonine-protein phosphatases, the SIT4 phosphatase-associated protein family, the TAP42 / TAP46-like superfamily, dual-specificity lipid and protein phosphatases, polynucleotide kinase 3 phosphatases, 5'-deoxynucleotidases, myosin family, histidine phosphatase superfamily, S-2-haloalkanoic acid dehalogenase, phosphatidylglycerol phosphatase, and haloic acid dehalogenase (HAD) hydrolase superfamily. ), phosphatase family (PA-phosphatase)Related phosphoesterase family, squalene / phytoene synthase family, terpene cyclase family, phosphatase App1, protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), halogen dehalogenase (HAD), sugar phosphophosphate phosphatase (HAD-like), polyphosphoinositide phosphatase (Fig4-like), casein kinase 1 (Se / Thr protein kinase), lipoprotein family, PPZ / Ppq1 family, clade-1 family, HAD-IA family hydrolases (HAD-IA family). IA hydrolase family), HAD-IF subfamily (IF family), dolichyldiphosphatase family, or haloperoxidase superfamily. In some embodiments, the phosphatase is phosphatidylphosphatase (APP1), polyacyl-2-phosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), inositol polyphosphate phosphatase (FIG4), 1-glycerol phosphate phosphatase 1 (GPP1), casein kinase I (HRR25), phosphatidylphosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), serine / threonine protein phosphatase (PTC3), serine / threonine protein phosphatase (PTC3), SIT4-associated protein SAP155, SIT4-associated protein SAP185, 2A Phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase (TEP1), polynucleotides3'-phosphatase (TPP1), phosphorylhydrolase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphorylhydrolase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), phosphatidylglycerol phosphatase (PgpA), phosphatidylglycerol phosphatase (PgpB), phosphatidylglycerol phosphatase (PgpC), carbapenyl diphosphatase (YbjG), dihydroxy diphosphatase (DOLPP1), phosphorylhydrolase (PLPP6), farnesyl diphosphatase (YisP), farnesol synthase (TPS2), acyclic sesquiterpene synthase (TPS1), or farnesol synthase (TPS13).

[0040] In some embodiments, the phosphatase is a phosphatase derived from a microorganism or a functional variant thereof, said microorganism including bacteria, yeast, or fungi, said bacteria being selected from Escherichia or Bacillus, said yeast being selected from Saccharomyces, or said phosphatase is derived from plants or humans (…). Homo sapiens Phosphohydrolases or functional variants thereof; in some embodiments, the phosphohydrolase is a phosphohydrolase or a functional variant thereof derived from bacteria, yeast or fungi, wherein the bacteria are selected from *Escherichia coli* (…). Escherichia coli Bacillus subtilis ( Bacillus subtilis The yeast is brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), or the phosphorylase is derived from plants or humans ( Man wise Phosphorylase or a functional variant thereof, wherein the plant is selected from moso bamboo ( Phyllostachys edulis ),corn( Corn ) or japonica rice ( Oryza sativa subsp. japonica In some embodiments, the phosphorylase is derived from *Saccharomyces cerevisiae* (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), Escherichia coli ( Escherichia coli ), human beings ( Homo sapiens Bacillus subtilis ( Bacillus subtilis ),bamboo( Phyllostachys edulis ),corn( Corn ) or japonica rice ( Oryza sativa subsp. japonica Acid hydrolases or their functional variants.

[0041] In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 11-226, or an amino acid sequence that is at least 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% identical to the amino acid sequence shown in SEQ ID NO. 11-226; in some embodiments, the phosphatase comprises an amino acid sequence selected from SEQ ID NO. 12, SEQ ID NO. 17, SEQ ID NO. 20, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 45, SEQ ID NO. 58, SEQ ID NO. 63, SEQ ID NO. 95, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19 ... NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID The amino acid sequence of NO.226, or selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160. SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID The amino acid sequence of NO. 226 has at least 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% sequence identity.

[0042] In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from those shown in SEQ ID NO. 237-452, or by a nucleotide sequence selected from those with at least 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% sequence identity with the nucleotide sequence shown in SEQ ID NO. 237-452; in some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 238, SEQ ID NO. 243, SEQ ID NO. 246, SEQ ID NO. 262, SEQ ID NO. 264, SEQ ID NO. 271, SEQ ID NO. 284, SEQ ID NO. 238, SEQ ID NO. 243, SEQ ID NO. 246, SEQ ID NO. 262, SEQ ID NO. 264, SEQ ID NO. 271, SEQ ID NO. 284, SEQ ID NO. 237-452. NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID The nucleotide sequence of NO.450, SEQ ID NO.451, SEQ ID NO.452, or the nucleotide sequence selected from SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO.271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383. SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ IDNO. 452 encodes a nucleotide sequence with at least 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% sequence identity.

[0043] In some embodiments, the first microorganism contains genes for the lysine pyrophosphate (LPP) synthesis pathway.

[0044] In some embodiments, the lysine pyrophosphate (LPP) synthesis pathway gene includes a farnesyl pyrophosphate (FPP) synthesis pathway gene, a geraniol geraniol pyrophosphate synthase (GGPPS) gene, and / or a lysine pyrophosphate synthase (LPPS) gene.

[0045] In some embodiments, the farnesyl pyrophosphate (FPP) synthesis pathway genes include one or more of the following: acetyl-CoA transferase gene, 3-methyl-3-hydroxyglutaryl-CoA synthase gene, 3-hydroxy-3-methylglutaryl-CoA reductase gene, mevalonate kinase gene, mevalonate-5-phosphate kinase gene, mevalonate-5-pyrophosphate decarboxylase gene, pentene pyrophosphate isomerase gene, or farnesyl pyrophosphate synthase gene.

[0046] In some embodiments, the first microorganism further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the activity of geraniol geraniol pyrophosphate synthase (GGPPS); and (2) At least one gene modification that enhances the activity of lysine pyrophosphate diol ester synthase (LPPS).

[0047] In some embodiments, at least one gene modification enhancing the activity of gerany-gerany-pyrophosphate synthase (GGPPS) includes: introducing an exogenous gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, increasing the gene copy number of the gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, and / or replacing the natural promoter of the endogenous gerany-gerany pyrophosphate synthase (GGPPS) with a promoter having a higher expression level; and / or At least one gene modification that enhances the activity of lysandrindiol pyrophosphate synthase (LPPS) includes: introducing an exogenous lysandrindiol pyrophosphate synthase (LPPS) gene, increasing the gene copy number of the lysandrindiol pyrophosphate synthase (LPPS) gene, and / or replacing the natural promoter of the endogenous lysandrindiol pyrophosphate synthase (LPPS) with a promoter that has a higher expression level.

[0048] In some embodiments, the exogenous geraniol geraniol pyrophosphate synthase (GGPPS) comprises the amino acid sequence shown in SEQ ID NO. 9, preferably, the exogenous geraniol geraniol pyrophosphate synthase (GGPPS) is encoded by the nucleotide sequence of SEQ ID NO. 235; and / or The exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a truncated lysine pyrophosphate diol ester synthase (LPPS); preferably, the truncated lysine pyrophosphate diol ester synthase (LPPS) is a signal peptide with the first 63 amino acids removed; preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) comprises the amino acid sequence shown in SEQ ID NO. 10; more preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) is encoded by the nucleotide sequence of SEQ ID NO. 236.

[0049] In some embodiments, the first microorganism further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the function of acetyl-CoA transferase; (2) At least one gene modification that enhances the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase; (4) At least one gene modification that enhances the activity of mevalonate kinase; (5) At least one gene modification that enhances the activity of mevalonate-5-phosphate kinase; (6) At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase; (7) At least one gene modification that enhances the activity of pentylene pyrophosphate isomerase; and (8) At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase.

[0050] In some implementations, at least one gene modification that enhances the function of acetyl-CoA transferase includes: introducing an exogenous acetyl-CoA transferase gene, increasing the gene copy number of the acetyl-CoA transferase gene, and / or replacing the natural promoter of endogenous acetyl-CoA transferase with a promoter that has a higher expression level. At least one gene modification that enhances the function of 3-methyl-3-hydroxyglutaryl-CoA synthase includes: introducing an exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene, increasing the gene copy number of the 3-methyl-3-hydroxyglutaryl-CoA synthase gene, and / or replacing the natural promoter of endogenous 3-methyl-3-hydroxyglutaryl-CoA synthase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase includes: introducing an exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene, increasing the gene copy number of the 3-hydroxy-3-methylglutaryl-CoA reductase gene, and / or replacing the natural promoter of endogenous 3-hydroxy-3-methylglutaryl-CoA reductase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of mevalonate kinase includes: introducing an exogenous mevalonate kinase gene, increasing the gene copy number of the mevalonate kinase gene, and / or replacing the natural promoter of endogenous mevalonate kinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-phosphokinase includes: introducing an exogenous mevalonate-5-phosphokinase gene, increasing the gene copy number of the mevalonate-5-phosphokinase gene, and / or replacing the natural promoter of endogenous mevalonate-5-phosphokinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase includes: introducing an exogenous mevalonate-5-pyrophosphate decarboxylase gene, increasing the gene copy number of the mevalonate-5-pyrophosphate decarboxylase gene, and / or replacing the natural promoter of endogenous mevalonate-5-pyrophosphate decarboxylase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of pentyrene pyrophosphate isomerase includes: introducing an exogenous pentyrene pyrophosphate isomerase gene, increasing the copy number of the pentyrene pyrophosphate isomerase gene, and / or replacing the natural promoter of the endogenous pentyrene pyrophosphate isomerase with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase includes: introducing an exogenous farnesyl pyrophosphate synthase gene, increasing the gene copy number of the farnesyl pyrophosphate synthase gene, and / or replacing the natural promoter of endogenous farnesyl pyrophosphate synthase with a promoter that has a higher expression level.

[0051] In some implementation schemes, wherein: The exogenous acetyl-CoA transferase comprises the amino acid sequence shown in SEQ ID NO.3, preferably, the exogenous acetyl-CoA transferase is encoded by the nucleotide sequence of SEQ ID NO.229; The exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase comprises the amino acid sequence shown in SEQ ID NO.4, preferably, the exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase is encoded by the nucleotide sequence of SEQ ID NO.230; The exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a truncated 3-hydroxy-3-methylglutaryl-CoA reductase, wherein the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is a truncated 3-hydroxy-3-methylglutaryl-CoA reductase with 528 amino acids removed from its N-terminus; preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase comprises the amino acid sequence shown in SEQ ID NO. 2, and preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is encoded by the nucleotide sequence of SEQ ID NO. 228; The exogenous mevalonate kinase comprises the amino acid sequence shown in SEQ ID NO.5, preferably, the exogenous mevalonate kinase is encoded by the nucleotide sequence of SEQ ID NO.231; The exogenous mevalonate-5-phosphokinase comprises the amino acid sequence shown in SEQ ID NO. 6, preferably, the exogenous mevalonate-5-phosphokinase is encoded by the nucleotide sequence of SEQ ID NO. 232; The exogenous mevalonate-5-pyrophosphate decarboxylase comprises the amino acid sequence shown in SEQ ID NO.7, preferably, the exogenous mevalonate-5-pyrophosphate decarboxylase is encoded by the nucleotide sequence of SEQ ID NO.233; The exogenous pentene pyrophosphate isomerase comprises the amino acid sequence shown in SEQ ID NO. 8, preferably, the exogenous pentene pyrophosphate isomerase is encoded by the nucleotide sequence of SEQ ID NO. 234; and / or The exogenous farnesyl pyrophosphate synthase comprises the amino acid sequence shown in SEQ ID NO.1, preferably encoded by the nucleotide sequence of SEQ ID NO.227.

[0052] In some embodiments, the first microorganism is a bacterium or yeast; preferably, the bacterium is selected from Escherichia, Corynebacterium, or Bacillus, and the yeast is selected from Saccharomyces, Pichia, Hansenula, Kluyveromyces, Phaffia, Schizosaccharomyces, Candida, Yarrowia, Hyphozyma, or Cryptococcus; more preferably, the bacterium is selected from Escherichia coli. Escherichia coli ), Corynebacterium glutamicum ( Corynebacterium glutamicum ), or Bacillus subtilis ( Bacillus subtilis The yeast is selected from brewer's yeast (Saccharomyces cerevisiae). Saccharomyces yeast Pichia pastoris () Shepherd's pie ), Phaffia colomata ( Komagataellaphaffii) , Saccharomyces cerevisiae ( Schizosaccharomyces pombe ), Candida albicans ( Candida albicans ), Candida utilis ( Useful Candida ), Yarrowia lipolytica ( Yarrow lipolytic ), Hansenula polymorpha ( Hansenula polymorpha Pichia pastoris (Canada) Peach Canadian Kluyveromyces marxianus and Kluyveromyces lactis. Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus white ) or Red Pavlova yeast ( Phaffia rhodozyma ).

[0053] Another aspect of the invention provides the use of any of the foregoing compositions in the production of perillaldehyde and / or perillyl lactone.

[0054] Another aspect of the present invention provides the use of the fungus or its derivatives with accession number ATCC 20918 in the production of perillaldehyde and / or perillyl lactone from lysine-1-benzenediol. Attached Figure Description

[0055] Figure 1 Metabolic diagram of the biosynthetic pathway of lysine diol in Saccharomyces cerevisiae, as an example.

[0056] Figure 2 The example reaction formula is shown for the production of lysine diol from lysine pyrophosphate via phosphorylase catalysis.

[0057] Figure 3 Design diagram for DNA donor of phosphorylase.

[0058] Figure 4 This is the gas chromatogram of lysine diol standard.

[0059] Figure 5 This is a gas chromatogram of the product after processing the sample of strain YC6.

[0060] Figure 6 This is the mass spectrum of the YC6 strain sample after treatment.

[0061] Figure 7 To produce perillaldehyde or perillyl alcohol by culturing fungus ATCC 20624 alone.

[0062] Figure 8 To produce perilla lactone by culturing fungus ATCC 20918 alone.

[0063] Figure 9 The effect of temperature on the products of mixed bacterial co-culture. Detailed Implementation

[0064] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0065] All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety. In case of conflict, this specification (including definitions) shall prevail. Furthermore, the materials, methods, and examples described herein are illustrative only and not intended to be restrictive.

[0066] When the terms “about” and “approximately” are used with numerical variables, they generally mean that the value of the variable and all values ​​of the variable are within the measurement or experimental error (e.g., the 95% confidence interval of the mean) or within a wider range of specified values ​​(e.g., ±5% or ±10%).

[0067] The term "comprising," or its variations such as "containing," "having," or "including," means to include the stated steps or elements, but does not exclude any other steps or elements. "Constitutes of," means to exclude steps or elements not listed. "Substantially constitutes of," means to include steps or elements that do not substantially affect the fundamental and novel features of the protected invention. The term "comprising" and its variations also include the cases of "consisting of specific steps or elements" and "substantially constitutes specific steps or elements."

[0068] When referring to a numerical range, it should be understood that the specific values ​​of its upper and lower limits are disclosed, as well as all intermediate ranges included therein, such as the intermediate range between its upper or lower limit and any intermediate value, or the intermediate range between any two intermediate values. Furthermore, any intermediate ranges, subranges, and all individual numerical values ​​described in the numerical range can be excluded from the numerical range.

[0069] The term “and / or” should be understood as any one of the multiple elements connected by the term, or a combination of any number of elements.

[0070] The inventors have discovered a biotransformation method for converting lysine-3-diol into perillaldehyde and / or perillaldehyde lactone, and have applied this biotransformation method to the production of perillaldehyde and / or perillaldehyde lactone, thereby providing a method and composition for producing perillaldehyde and / or perillaldehyde lactone. The method and composition for producing perillaldehyde and / or perillaldehyde lactone provided by this invention have high production efficiency and yield.

[0071] The method for producing perillaldehyde and / or perillyl lactone provided by the present invention includes: The following components are contacted: component (1) and component (2) are brought together to produce perillaldehyde and / or perillyl lactone; wherein, The first component is selected from components containing lysine diol. The second component is selected from enzymes or microorganisms or their cell lysates or enzyme extracts that can convert lysine diol into perillyl diol and / or perillyl lactone.

[0072] In this document, the term "Labdenediol (LOH)" includes (E)-Labdenediol-13-en-8,15-diol. For example, the term "Labdenediol (LOH)" can include compounds with the chemical name (1R,2R,4aS,8aS)-1-[(E)-5-hydroxy-3-methylpentan-3-enyl]-2,5,5,8a-tetramethyl-3,4,4a,6,7,8-hexahydro-1H-naphth-2-ol] with the molecular formula C2 20 H 36 O2, CAS No. 10267-31-9.

[0073] The term "contact" refers to two or more components being physically close enough for a reaction (e.g., biocatalysis or biotransformation) to occur involving the two or more components. In some embodiments, the contact between component (1) and component (2) is sufficient to allow component (1) to be converted into stilbene glycol and / or stilbene lactone by the action of component (2). In some embodiments, the term "contact" may refer to a mixture of two or more components. When the two or more components contain an organism, such as a microbial cell, "contact" includes mixing and incubating, or mixing and culturing, the two or more components.

[0074] The term "enzyme or microorganism capable of converting lysenoside diol to perillyl diol and / or perillyl lactone" refers to any enzyme or microorganism capable of converting lysenoside diol to perillyl diol and / or perillyl lactone. Such microorganisms can include those naturally possessing this ability within their cells, such as microorganisms naturally possessing enzymes capable of converting lysenoside diol to perillyl diol and / or perillyl lactone, or those possessing or enhancing this ability through genetic modification, such as introducing an exogenous enzyme gene capable of converting lysenoside diol to perillyl diol and / or perillyl lactone through genetic modification, or microorganisms whose activity in cells is enhanced by genetic modification of the enzyme capable of converting lysenoside diol to perillyl diol and / or perillyl lactone. "Microorganism capable of converting lysenoside diol to perillyl diol and / or perillyl lactone" also includes microorganisms that acquire the ability to convert lysenoside diol to perillyl diol and / or perillyl lactone through artificial mutagenesis, radiation, or other methods.

[0075] It should be understood that, according to the biotransformation method disclosed in this invention, those skilled in the art can screen for enzymes or microorganisms capable of converting lysine-enriched diol to perillyl glycol and / or perillyl lactone, for example, by contacting candidate enzymes or microorganisms with lysine-enriched diol and detecting whether perillyl glycol and / or perillyl lactone are produced. In some embodiments, cell lysates and / or enzyme extracts of enzymes or microorganisms capable of converting lysine-enriched diol to perillyl glycol and / or perillyl lactone are capable of converting lysine-enriched diol to perillyl glycol and / or perillyl lactone, or contain enzymes capable of converting lysine-enriched diol to perillyl glycol and / or perillyl lactone. In some embodiments, this can be determined, for example, by contacting the cell lysate or enzyme extract with lysine-enriched diol and detecting whether perillyl glycol and / or perillyl lactone are produced.

[0076] In some embodiments, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from... Hyphozyma roseoniger Or Cryptococcus lightensis ( Cryptococcus albidus In some embodiments, the microorganisms capable of converting lysine-3-diol into perillylene-3-diol and / or perillylene lactone are selected from fungi with accession number ATCC 20624, fungi with accession number ATCC20918, or derivatives thereof.

[0077] The fungus with accession number ATCC 20624 is disclosed in US4798799A (the full text of which is incorporated herein by reference), which is Hyphozyma roseoniger This fungus is deposited at the American Type Culture Collection (ATCC) under accession number ATCC 20624 and is available for purchase from the ATCC. It can convert labdenediol or lysine into labdenediol and / or labdenolactone.

[0078] The fungus with accession number ATCC 20918 is disclosed in US4970163A, US5155029A, and US5212078A (the full text of which is incorporated herein by reference), and is *Cryptococcus faecium*. Cryptococcus albidus This fungus, deposited at the American Type Culture Collection (ATCC) with accession number ATCC20624, can be purchased from the ATCC. It can convert perillyl alcohol to perillyl lactone.

[0079] "Derived bacteria" refers to microorganisms that share the same taxonomic characteristics as the parent microorganism, such as having the same 16S rDNA, 18S rDNA, and / or ITS, and possessing similar (or enhanced) functions or activities in converting lysine-3-diol to perillylene-2-diol and / or perillylene lactone. Derived bacteria may exhibit certain changes in their genomic DNA relative to the parent microorganism, which may alter one or more traits of the derived bacteria but do not result in a change of species. In some embodiments, the changes in the genomic DNA of the derived bacteria relative to the parent microorganism may not exceed 1 gene, 2 genes, 3 genes, 4 genes, 5 genes, 6 genes, 7 genes, 8 genes, 9 genes, 10 genes, 11 genes, 12 genes, 15 genes, 20 genes, 25 genes, 30 genes, 35 genes, 40 genes, 45 genes, or 50 genes. In some implementations, the variation in the genomic DNA of the derived bacteria relative to the parent microorganism may not exceed 0.001%, 0.005%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, or 2% (calculated by base pairs). The derived bacteria may be obtained by genetically modifying the parent microorganism, or by artificial mutagenesis, radiation, or other methods.

[0080] In the methods described herein, lysine diol may be provided in any suitable manner, such as in the form of an isolated compound, or in the form of a mixture of lysine diol with any other substance, or in the form of a microorganism capable of producing lysine diol, or in the form of a substance containing lysine diol obtained from the microorganism (e.g., culture medium, supernatant, cell lysate, or extract).

[0081] In some embodiments, the isolated lysine diol contains at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% lysine diol by weight.

[0082] The term "microorganism capable of producing lysenoside diol" refers to any microorganism capable of producing lysenoside diol, including those with this ability naturally present in their cells, such as those with naturally occurring lysenoside diol synthesis pathways, or those that have been genetically modified to possess or enhance this ability. For example, genetic modification can introduce one or more exogenous enzyme genes to create a lysenoside diol synthesis pathway in the cell, or genetic modification can enhance one or more enzyme genes in the lysenoside diol synthesis pathway in the cell, thereby increasing the production of lysenoside diol. "Microorganism capable of producing lysenoside diol" also includes microorganisms that acquire the ability to produce lysenoside diol through artificial mutagenesis, radiation, or other methods.

[0083] In this article, the supernatant of "microorganisms capable of producing lysine diol" can be a supernatant obtained without lysing cells, such as the supernatant obtained after centrifuging or allowing the cell culture to stand, or a supernatant obtained by lysing cells, such as the supernatant obtained after centrifuging or allowing the cells to stand after lysing.

[0084] In this article, the extract of "microorganisms capable of producing lysinediol" refers to fractions containing lysinediol extracted from the cells, cell cultures, culture media, supernatants, etc. of such microorganisms.

[0085] In some embodiments, the microorganisms capable of producing lysine diol are recombinant engineered bacteria. In some embodiments, the recombinant engineered bacteria introduces from an exogenous source the genes for one or more enzymes in the lysine diol synthesis pathway, the genes for which may be contained in a free expression vector introduced into the host bacterium or integrated into the host bacterium's chromosome.

[0086] In some embodiments, the method is used to produce perillaldehyde, or perillaldehyde and perillaldehyde, wherein the microorganism capable of converting lysine-enriched diol to perillaldehyde and / or perillaldehyde can be a microorganism capable of converting lysine-enriched diol to perillaldehyde, or to perillaldehyde and perillaldehyde. In some embodiments, the microorganism is selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganism capable of converting lysine-enriched diol to perillaldehyde and / or perillaldehyde is selected from... Hyphozyma rose-colored Or Cryptococcus albidus. In some embodiments, the microorganism is selected from fungi with accession number ATCC 20624, fungi with accession number ATCC 20918, or derivatives thereof.

[0087] In some embodiments, the method is used to produce perillaldehyde, wherein the microorganism capable of converting lysine-enriched diol to perillaldehyde and / or perillaldehyde lactone may be a microorganism capable of converting lysine-enriched diol to perillaldehyde. In some embodiments, the microorganism is selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganism capable of converting lysine-enriched diol to perillaldehyde is selected from *Cryptococcus faecium*. Cryptococcus albidus The microorganism is selected from fungi with accession number ATCC 20918 or their derivatives. In some embodiments, the microorganism is selected from fungi with accession number ATCC 20918 or their derivatives.

[0088] In some embodiments, the method is used to produce perillaldehyde and / or perillyl lactone, wherein the first component is selected from microorganisms capable of producing lysenoside diol. In some embodiments, the microorganisms capable of producing lysenoside diol are recombinant engineered bacteria. In some embodiments, the second component is a microorganism capable of converting lysenoside diol into perillaldehyde diol and / or perillyl lactone. In some embodiments, the microorganism capable of converting lysenoside diol into perillaldehyde diol and / or perillyl lactone is selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganism capable of converting lysenoside diol into perillaldehyde diol and / or perillyl lactone is selected from... Hyphozyma roseoniger Or Cryptococcus lightensis ( Cryptococcus albidus In some embodiments, the microorganism is selected from fungi with accession number ATCC 20624, fungi with accession number ATCC 20918, or derivatives thereof.

[0089] The present invention also provides a composition comprising the following two microorganisms: The first microorganism is selected from microorganisms capable of producing lysine diol (e.g., recombinant engineered bacteria), such as those described above; The second microorganism is selected from those capable of converting lysine-enrichediol into perillyl alcohol and / or perillyl lactone, for example, as described above.

[0090] Wild-type Escherichia coli or yeast and other microorganisms can use glucose, ethanol, etc., as carbon sources to catalyze a multi-step reaction, catalyzing the formation of sclareol via the intermediate lysenoside pyrophosphate (LPP). This synthesis involves the mevalonate (MVA) pathway and / or the isoprene (PDP) pathway. The inventors have discovered that lysenoside pyrophosphate (LPP) can also serve as a precursor to the target product lysenoside diol. A specific phosphorylase can catalyze the formation of lysenoside pyrophosphate (LPP) to lysenoside diol (LOH). The inventors have screened phosphorylases that can efficiently catalyze the dephosphatemization of lysenoside pyrophosphate (LPP) to produce lysenoside diol (LOH).

[0091] In some embodiments, the recombinant engineered bacteria described herein capable of producing lysinediol contains a phosphorylase that catalyzes the production of lysinediol pyrophosphate (LPP) from lysinediol.

[0092] In some embodiments, the phosphatase may be derived from Mg(2+)-dependent phosphatidate phosphatase, phosphatidicacid phosphatase type 2, serine / threonine-protein phosphatase, HAD-like hydrolase superfamily, phosphate (PA) phosphatase, polyphosphoinositide phosphatase, serine / threonine-protein kinases, alkaline phosphatase, phosphoprotein phosphatase family, serine / threonine-protein phosphatases, or the SIT4 phospholipase-associated protein family. The following are listed as superfamilies in the phosphatase family: phosphatase-associated protein family, TAP42 / TAP46-like superfamily, dual-specificity lipid and protein phosphatase, polynucleotide kinase 3 phosphatase, 5'-deoxynucleotidase, myosin family, histidine phosphatase superfamily, S-2-haloalkanoic acid dehalogenase, phosphatidylglycerol phosphatase, HAD-hydrolase superfamily, PA-phosphatase related phosphoesterase family, and squalene / phytoene synthase family.Syntheses, terpene cyclases, phosphatase App1, protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), halogen dehalogenases (HAD), sugar phosphophosphate phosphatase (HAD-like), polyphosphoinositide phosphatase (Fig4-like), casein kinase 1 (Serine / Thr protein kinase), lipoprotein family, PPZ / Ppq1 family, clade-1 family, HAD-IA hydrolase family, and HAD-IF subfamily (IF). The phosphatase family, dolichoyldiphosphatase family, or haloperoxidase superfamily; preferably, the phosphatase is phosphatidylate phosphatase (APP1), dolichoyldiphosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), inositol polyphosphate phosphatase (FIG4), 1-glycerol phosphate phosphatase 1 (GPP1), casein kinase I (HRR25), phosphatidylate phosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), serine / threonine protein phosphatase (PTC3), serine / threonine protein phosphatase (PTC3), SIT4 Associated proteins SAP155, SIT4 associated protein SAP185, class 2A phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase (TEP1), and polynucleotides3'-phosphatase (TPP1), phosphorylhydrolase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphorylhydrolase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), phosphatidylglycerol phosphatase (PgpA), phosphatidylglycerol phosphatase (PgpB), phosphatidylglycerol phosphatase (PgpC), carbapenyl diphosphatase (YbjG), dihydroxy diphosphatase (DOLPP1), phosphorylhydrolase (PLPP6), farnesyl diphosphatase (YisP), farnesol synthase (TPS2), acyclic sesquiterpene synthase (TPS1), or farnesol synthase (TPS13).

[0093] In some embodiments, the phosphatase is a phosphatase derived from a microorganism or a functional variant thereof, said microorganism including bacteria, yeast, or fungi, said bacteria being selected from Escherichia or Bacillus, said yeast being selected from Saccharomyces, or said phosphatase is derived from plants or humans (…). Homo sapiens Phosphohydrolases or their functional variants thereof; preferably, the phosphohydrolase is a phosphohydrolase or its functional variant derived from bacteria, yeast or fungi, wherein the bacteria are selected from *Escherichia coli* (…). Escherichia coli Bacillus subtilis ( Bacillus subtilis The yeast is brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), or the phosphorylase is derived from plants or humans ( Homo sapiens Phosphorylase or a functional variant thereof, wherein the plant is selected from moso bamboo ( Phyllostachys edulis ),corn( Zea corn ) or japonica rice ( Oryza sativa subsp. japonica More preferably, the phosphorylase may be derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), Escherichia coli ( Escherichia coli ), human beings ( Man wise Bacillus subtilis ( Bacillus subtilis ),bamboo( Phyllostachys edulis ),corn( Zea corn ) or japonica rice ( Oryza sativa subsp. japonica Phosphohydrolases or their functional variants.

[0094] In some embodiments, the phosphorylase may be derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces yeastPhosphatidyl phosphatase (APP1), polyacyl diphosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), polyphosphatidylinositol phosphatase (FIG4), 1-glycerophosphate phosphohydrolase 1 (GPP1), casein kinase I (HRR25), phosphatidyl phosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), serine / threonine protein phosphatase (PTC3), serine / threonine protein phosphatase (PTC3), SIT4-associated protein SAP155, SIT4-associated protein SAP185, 2A Phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase (TEP1), polynucleotide 3'-phosphatase (TPP1), phosphatase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphatase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), or functional variants thereof. In some embodiments, the phosphatase comprises an amino acid sequence selected from SEQ ID NO. 11-216 or a functional variant thereof.

[0095] In some embodiments, the phosphorylase may be derived from *Escherichia coli* (…). Escherichia coli Phosphatidylglycerol phosphatase (PgpA) or a functional variant thereof. In some embodiments, the phosphatase may be derived from *Escherichia coli* (PgpA). Escherichia coli Phosphatidylglycerol phosphatase (PgpA) of K12 or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 217 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 443.

[0096] In some embodiments, the phosphorylase may be derived from *Escherichia coli* (…). Escherichia coli Phosphatidylglycerol phosphatase (PgpB) or a functional variant thereof. In some embodiments, the phosphatase may be derived from *Escherichia coli* (PgpB). Escherichia coliPhosphatidylglycerol phosphatase (PgpB) of K12 or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 218 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 444.

[0097] In some embodiments, the phosphorylase may be derived from *Escherichia coli* (…). Escherichia coli Phosphatidylglycerol phosphatase (PgpC) or a functional variant thereof. In some embodiments, the phosphatase may be derived from *Escherichia coli* (PgpC). Escherichia coli Phosphatidylglycerol phosphatase (PgpC) of K12 or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 219 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 445.

[0098] In some embodiments, the phosphorylase may be derived from *Escherichia coli* (…). Escherichia coli The enzyme is a carbapenyl diphosphatase (YbjG) or a functional variant thereof. In some embodiments, the phosphatase may be derived from *Escherichia coli* (YbjG). Escherichia coli The phosphatase is a carbapenyl diphosphatase (YbjG) of K12 or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 220 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 446.

[0099] In some embodiments, the phosphorylase may be derived from human ( Homo sapiens The phosphatase is a dihydroxy diphosphatase (DOLPP1) or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 221 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 447.

[0100] In some embodiments, the phosphorylase may be derived from human ( Homo sapiens The phosphorylase (PLPP6) or a functional variant thereof. In some embodiments, the phosphorylase comprises an amino acid sequence as shown in SEQ ID NO. 222 or a functional variant thereof. In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 448.

[0101] In some embodiments, the phosphorylase may be derived from Bacillus subtilis (Bacillus subtilis) Bacillus subtle The enzyme is a farnesyl diphosphatase (YisP) or a functional variant thereof. In some embodiments, the phosphatase comprises an amino acid sequence as shown in SEQ ID NO. 223 or a functional variant thereof. In some embodiments, the phosphatase is encoded by a nucleotide sequence selected from SEQ ID NO. 449.

[0102] In some embodiments, the phosphorylase may be derived from moso bamboo (… Phyllostachys edulis The phosphorylase is farnesol synthase (TPS2) or a functional variant thereof. In some embodiments, the phosphorylase comprises an amino acid sequence as shown in SEQ ID NO. 224 or a functional variant thereof. In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 450.

[0103] In some embodiments, the phosphorylase may be derived from corn ( Corn The noncyclic sesquiterpene synthase (TPS1) or a functional variant thereof. In some embodiments, the phosphorylase comprises an amino acid sequence as shown in SEQ ID NO. 225 or a functional variant thereof. In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 451.

[0104] In some embodiments, the phosphorylase may be derived from japonica rice ( Oryza sativa subsp. Japanese) The enzyme is farnesol synthase (TPS13) or a functional variant thereof. In some embodiments, the phosphorylase comprises an amino acid sequence as shown in SEQ ID NO. 226 or a functional variant thereof. In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 452.

[0105] In this invention, through exemplary catalytic experiments of lysandrindiol pyrophosphate (LPP) to lysandrindiol (LOH) and comparative analysis of sequence results of phosphorylases from different sources, it was found that there is approximately 80% homology difference among phosphorylases from different sources.

[0106] In some embodiments, the phosphatase comprises an amino acid sequence selected from SEQ ID NO. 11-226, or an amino acid sequence having at least 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% sequence identity with an amino acid sequence selected from SEQ ID NO. 11-226. Preferably, the phosphatase comprises an amino acid sequence selected from SEQ ID NO. 11-226.

[0107] In some embodiments, the phosphorylase comprises an enzyme selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO. The amino acid sequences of SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, and SEQ ID NO.226.Or selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ The amino acid sequences of SEQ ID NO. 222, SEQ ID NO. 223, SEQ ID NO. 224, SEQ ID NO. 225, and SEQ ID NO. 226 have amino acid sequences with at least 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% sequence identity. The phosphohydrolases of the above amino acid sequences can effectively catalyze the formation of lysandrindiol pyrophosphate (LPP) to lysandrindiol (LOH).

[0108] In some embodiments, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 237-452, or by a nucleotide sequence having at least 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% sequence identity with a nucleotide sequence selected from SEQ ID NO. 237-452. Preferably, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 237-452. In some embodiments, the gene encoding the phosphorylase is codon-optimized for expression in a host bacterium (e.g., Saccharomyces cerevisiae). In some embodiments, the phosphohydrolase is encoded by a nucleotide sequence selected from SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO. 271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ ID NO.452, or by a sequence that is homologous to a sequence selected from SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO. 271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.Nucleotide sequences 449, SEQ ID NO. 450, SEQ ID NO. 451, and SEQ ID NO. 452 have at least 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% sequence identity. The phosphorylase encoded by the above nucleotide sequences can efficiently catalyze the conversion of lysandrindiol pyrophosphate (LPP) to lysandrindiol (LOH).

[0109] In this invention, the term "recombinant engineered bacteria containing the phosphatase" means that the recombinant engineered bacteria express the phosphatase, or that it contains the coding gene for the phosphatase. The phosphatase can be endogenous or heterologous to the host bacteria. In some embodiments, the recombinant engineered bacteria are genetically modified to alter (typically increase) the expression and / or activity of the phosphatase in the recombinant engineered bacteria. For example, the gene for the phosphatase can be introduced exogenously into the recombinant engineered bacteria, or the copy number of the phosphatase gene naturally present in the recombinant engineered bacteria (i.e., endogenous) can be increased, and / or the natural promoter of the endogenous phosphatase gene naturally present in the recombinant engineered bacteria (i.e., endogenous) can be replaced with a promoter that has a higher expression level. In some embodiments, the recombinant engineered bacteria contain an exogenously introduced phosphatase gene. The exogenously introduced phosphatase gene can be contained in a free expression vector introduced into the host bacteria or integrated into the chromosome of the host bacteria.

[0110] In some embodiments, the recombinant engineered bacteria further includes a lysine pyrophosphate (LPP) synthesis pathway gene to produce lysine pyrophosphate in the recombinant engineered bacteria.

[0111] In some embodiments, the lysine pyrophosphate (LPP) synthesis pathway genes include farnesyl pyrophosphate (FPP) synthesis pathway genes, gerany-gerany-pyrophosphate synthase (GGPPS) genes, and / or lysine pyrophosphate synthase (LPPS) genes.

[0112] In some implementations, the "Farnesy pyrophosphate (FPP) synthesis pathway" includes the following enzymatically catalyzed reactions: (1) acetyl-coenzyme A (acetyl-CoA) is converted to acetoacetyl-CoA via acetyl-CoA transferase; (2) acetoacetyl-CoA is converted to 3-hydroxy-3-methylglutaryl-CoA via 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) mevalonic acid (MVA) is converted to 3-hydroxy-3-methylglutaryl-CoA via 3-hydroxy-3-methylglutaryl-CoA reductase; (4) (5) Mevalonate (MVA) catalyzed by mevalonate kinase to generate mevalonate phosphate (MVAP); (6) Mevalonate phosphate (MVAP) catalyzed by mevalonate-5-phosphate kinase to generate mevalonate pyrophosphate (MVAP); (7) Isopentyl pyrophosphate (IPP) catalyzed by mevalonate-5-pyrophosphate decarboxylase to generate dimethylallyl diphosphate (DMAPP), or Isopentyl pyrophosphate (DMAPP) catalyzed by mevalonate pyrophosphate isomerase to generate isopentenyl pyrophosphate (IPP); and (8) Isopentyl pyrophosphate (IPP) and / or dimethylallyl pyrophosphate (DMAPP) catalyzed by farnesyl pyrophosphate synthase to generate farnesyl diphosphate (FPP).

[0113] In some embodiments, the farnesyl pyrophosphate (FPP) synthesis pathway genes include one or more of the following: acetyl-CoA transferase gene, 3-methyl-3-hydroxyglutaryl-CoA synthase gene, 3-hydroxy-3-methylglutaryl-CoA reductase gene, mevalonate kinase gene, mevalonate-5-phosphate kinase gene, mevalonate-5-pyrophosphate decarboxylase gene, pentene pyrophosphate isomerase gene, or farnesyl pyrophosphate synthase gene.

[0114] In some implementations, the “lysine pyrophosphate diol ester (LPP) synthesis pathway gene” includes the mevalonic acid (MVA) pathway gene.

[0115] The mevalonate pathway (MVA) is a metabolic pathway that synthesizes isopentenyl pyrophosphate (IPP) and dimethylpropene pyrophosphate (DMAPP) from acetyl-CoA. It is present in all eukaryotes, bacteria, and many viruses. The products of this pathway can be considered as activated isoprene units, which are precursors for the synthesis of biomolecules such as steroids and terpenoids. Its synthetic pathway can include the following enzymatic reactions: (1) acetyl-CoA is converted to acetoacetyl-CoA via acetyl-CoA transferase; (2) acetoacetyl-CoA is converted to acetoacetyl-CoA via 3-methyl-3-hydroxyglutaryl-CoA synthase. (3) 3-hydroxy-3-methylglutaryl-CoA is generated by acetoacetyl-CoA; (4) 3-hydroxy-3-methylglutaryl-CoA is converted to mevalonic acid (MVA) by 3-hydroxy-3-methylglutaryl-CoA reductase; (5) mevalonic acid phosphate (MVAP) is generated by mevalonic acid phosphate (MVAP) by mevalonic acid kinase; (6) isopentenyl pyrophosphate (IPP) is generated by isopentenyl pyrophosphate (IPP) by isopentenyl pyrophosphate isomerase; (7) dimethylallyl diphosphate is generated by isopentenyl pyrophosphate (IPP) by isopentenyl pyrophosphate isomerase. DMAPP, or isopentenyl pyrophosphate (IPP) catalyzed by pentene pyrophosphate isomerase, is used to catalyze the synthesis of dimethylallyl diphosphate (DMAPP). This exemplary synthetic route is described herein as follows: Figure 1 As shown.

[0116] In some embodiments, the “lysandrindiol pyrophosphate (LPP) synthesis pathway gene” includes an enzyme gene for synthesizing an enzyme that can be catalyzed to produce lysandrindiol pyrophosphate (LPP) or its precursors. In some embodiments, the precursors may, for example, include farnesyl diphosphate (FPP) and / or geranylgeranyl diphosphate (GGPP). In some embodiments, the lysandrin diol pyrophosphate (LPP) synthesis pathway gene includes a farnesyl pyrophosphate synthase gene comprising the production of farnesyl diphosphate (FPP) from isopentenyl pyrophosphate (IPP) and / or dimethylpropene pyrophosphate (DMAPP), a geranylgeranyl pyrophosphate synthase (GGPPS) gene comprising the production of geranylgeranyl diphosphate (GGPP) from farnesyl diphosphate (FPP), and / or a lysandrin diol pyrophosphate synthase (LPPS) gene comprising the production of lysandrin diol pyrophosphate (LPP) from geranylgeranyl diphosphate (GGPP).

[0117] In some embodiments, the “lysine pyrophosphate diol ester (LPP) synthesis pathway” may include the following enzymatically catalyzed reactions: (1) catalysis of acetyl-coenzyme A (acetyl-CoA) by acetyl-CoA transferase to generate acetoacetyl-CoA; (2) catalysis of acetoacetyl-CoA by 3-methyl-3-hydroxyglutaryl-CoA synthase. (3) 3-hydroxy-3-methylglutaryl-CoA is generated by acetoacetyl-CoA; (4) 3-hydroxy-3-methylglutaryl-CoA is converted to mevalonic acid (MVA) by 3-hydroxy-3-methylglutaryl-CoA reductase; (5) mevalonic acid phosphate (MVAP) is generated by mevalonic acid phosphate (MVAP) by mevalonic acid kinase; (6) isopentenyl pyrophosphate (IPP) is generated by isopentenyl pyrophosphate (IPP) by isopentenyl pyrophosphate isomerase; (7) dimethylallyl pyrophosphate (IPP) is generated by isopentenyl pyrophosphate (IPP) by isopentenyl pyrophosphate isomerase. (7) dimethylallyl diphosphate (DMAPP) catalyzed by pentyle pyrophosphate isomerase to generate isopentenyl pyrophosphate (IPP); (8) isopentenyl diphosphate (IPP) and / or dimethylallyl pyrophosphate (DMAPP) catalyzed by farnesyl pyrophosphate synthase to generate farnesyl diphosphate (FPP); (9) farnesyl diphosphate (FPP) catalyzed by geranylgeranyl pyrophosphate synthase (BTS1) to generate geranylgeranyl diphosphate (GGPP); (10) geranylgeranyl diphosphate (GGPP) catalyzed by lysine pyrophosphate synthase (LPPS) to generate lysine pyrophosphate (LPP); this exemplary synthetic route is described herein as follows. Figure 1 As shown.

[0118] One or more, or all, of the genes in any of the synthetic pathways mentioned herein may be endogenous or exogenous. When the host bacterium naturally possesses the synthetic pathway, all genes in the synthetic pathway may be endogenous. In some embodiments, when the host bacterium does not naturally possess a certain synthetic pathway, for example, lacking one or more enzymes in any of the synthetic pathways mentioned herein, the missing one or more enzymes may be provided by introducing exogenous enzyme genes into the host bacterium to express the missing one or more enzymes in the host bacterium. In some embodiments, one or more, or all, of the genes in any of the synthetic pathways mentioned herein may be introduced exogenously into the host bacterium. In some embodiments, the exogenous gene may be contained in a free expression vector introduced into the host bacterium or integrated into the chromosome of the host bacterium.

[0119] In some embodiments, the recombinant engineered bacteria of the present invention may further include gene modifications that enhance the function of one or more enzymes in any of the synthetic pathways mentioned herein, in order to further improve the production efficiency and / or yield of the product lysine diol.

[0120] For one or more of the enzymes described above, gene modifications to enhance their function may include increasing the enzyme's activity and / or overexpressing the enzyme. Increasing the enzyme's activity may, for example, include introducing a foreign enzyme gene, which may be a foreign gene with a sequence different from the endogenous gene, particularly a foreign enzyme gene with higher enzyme activity. The foreign enzyme may be an enzyme derived from a species different from the engineered bacteria, or a mutant of the wild-type enzyme. Introducing a foreign gene may retain the presence of the endogenous gene with higher enzyme activity, or it may replace the endogenous gene with a foreign gene. Overexpressing the enzyme may, for example, include increasing the copy number of the endogenous enzyme gene, and / or replacing the enzyme's natural promoter with a promoter that has a higher expression level. In some embodiments, increasing the gene copy number of the enzyme includes introducing an additional exogenous enzyme gene into the engineered bacteria without altering the expression of the endogenous enzyme gene. The exogenous enzyme gene can be one, two, three, or more copies, and can have the same or different sequences as the endogenous enzyme. For example, the exogenous enzyme can originate from the same species as the engineered bacteria, or it can have higher enzyme activity compared to the endogenous enzyme, for example, it can be an enzyme from a different species than the engineered bacteria, or a mutant of the wild-type enzyme. The overexpression can also be achieved by modifying the regulatory sequence (e.g., the promoter) operatively linked to the gene encoding the enzyme in the engineered bacteria, for example, by replacing the original promoter (e.g., the promoter of the gene naturally present in the host cell) with a stronger promoter, or by introducing mutations or inserting new regulatory elements into the promoter region to enhance its activity. Those skilled in the art will understand that the term "stronger promoter" refers to a promoter that has stronger promoter activity relative to a pre-existing promoter in the host cell (e.g., a promoter of the gene naturally present in the host cell), such as a stronger ability to bind to the transcription initiation complex and / or a stronger RNA polymerase binding ability. As is known to those skilled in the art, using a stronger promoter can increase the expression level of a gene operatively linked to the promoter (e.g., increase the expression level of the protein encoded by that gene).

[0121] In some embodiments, the recombinant engineered bacteria of the present invention may further comprise one or more of the following gene modifications: (1) At least one gene modification that enhances the activity of geraniol geraniol pyrophosphate synthase (GGPPS); and (2) At least one gene modification that enhances the activity of lysine pyrophosphate diol ester synthase (LPPS). In some embodiments, at least one gene modification enhancing the activity of geraniol-geraniol pyrophosphate synthase (GGPPS) includes: introducing an exogenous geraniol-geraniol pyrophosphate synthase (GGPPS) gene, increasing the copy number of the geraniol-geraniol pyrophosphate synthase (GGPPS) gene, and / or replacing the natural promoter of the endogenous geraniol-geraniol pyrophosphate synthase (GGPPS) with a promoter having a higher expression level. In some embodiments, the exogenously introduced geraniol-geraniol pyrophosphate synthase (GGPPS) gene encodes a geraniol-geraniol pyrophosphate synthase (GGPPS) derived from *Saccharomyces cerevisiae* or a functional variant thereof. In some embodiments, the exogenous geraniol-geraniol pyrophosphate synthase (GGPPS) comprises the amino acid sequence shown in SEQ ID NO. 9. In some embodiments, the exogenous geraniol-geraniol pyrophosphate synthase (GGPPS) is encoded by the nucleotide sequence of SEQ ID NO. 235.

[0122] In some embodiments, at least one gene modification enhancing the activity of lysine pyrophosphate diol ester synthase (LPPS) includes: introducing an exogenous lysine pyrophosphate diol ester synthase (LPPS) gene, increasing the copy number of the LPPS gene, and / or replacing the natural promoter of the endogenous lysine pyrophosphate diol ester synthase (LPPS) with a promoter that has a higher expression level. In some embodiments, the introduced exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a gene derived from *Perilla frutescens* (a type of perilla frutescens). Clary sage The introduced exogenous lysine pyrophosphate diol ester synthase (LPPS) or a functional variant thereof. In some embodiments, the introduced exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a truncated lysine pyrophosphate diol ester synthase (LPPS). In some embodiments, the truncated lysine pyrophosphate diol ester synthase (LPPS) has a signal peptide with the first 63 amino acids removed. In some embodiments, the introduced exogenous lysine pyrophosphate diol ester synthase (LPPS) gene is codon-optimized for a host bacterium (e.g., Saccharomyces cerevisiae). In some embodiments, the introduced exogenous lysine pyrophosphate diol ester synthase (LPPS) gene contains the amino acid sequence shown in SEQ ID NO. 10. In some embodiments, the exogenous lysine pyrophosphate diol ester synthase (LPPS) is encoded by the nucleotide sequence of SEQ ID NO. 236.

[0123] In some embodiments, the recombinant engineered bacteria of the present invention may further comprise one or more of the following gene modifications: (1) At least one gene modification that enhances the function of acetyl-CoA transferase; (2) At least one gene modification that enhances the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase; (4) At least one gene modification that enhances the activity of mevalonate kinase; (5) At least one gene modification that enhances the activity of mevalonate-5-phosphate kinase; (6) At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase; (7) At least one gene modification that enhances the activity of pentylene pyrophosphate isomerase; and (8) At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase.

[0124] In some embodiments, at least one gene modification enhancing the function of acetyl-CoA transferase includes: introducing an exogenous acetyl-CoA transferase gene, increasing the copy number of the acetyl-CoA transferase gene, and / or replacing the natural promoter of the endogenous acetyl-CoA transferase with a promoter having a higher expression level. In some embodiments, the introduced exogenous acetyl-CoA transferase gene encodes an acetyl-CoA transferase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous acetyl-CoA transferase comprises the amino acid sequence shown in SEQ ID NO. 3, preferably, the exogenous acetyl-CoA transferase is encoded by the nucleotide sequence of SEQ ID NO. 229.

[0125] In some embodiments, at least one gene modification enhancing the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase includes: introducing an exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene, increasing the copy number of the 3-methyl-3-hydroxyglutaryl-CoA synthase gene, and / or replacing the natural promoter of the endogenous 3-methyl-3-hydroxyglutaryl-CoA synthase with a promoter having a higher expression level. In some embodiments, the introduced exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene encodes a 3-methyl-3-hydroxyglutaryl-CoA synthase derived from *Saccharomyces cerevisiae* or a functional variant thereof. In some embodiments, the introduced exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase comprises the amino acid sequence shown in SEQ ID NO. 4, preferably, the exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase is encoded by the nucleotide sequence of SEQ ID NO. 230.

[0126] In some embodiments, at least one gene modification enhancing the activity of 3-hydroxy-3-methylglutaryl-CoA reductase includes: introducing an exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene, increasing the copy number of the 3-hydroxy-3-methylglutaryl-CoA reductase gene, and / or replacing the natural promoter of the endogenous 3-hydroxy-3-methylglutaryl-CoA reductase with a promoter having a higher expression level. In some embodiments, the introduced exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a 3-hydroxy-3-methylglutaryl-CoA reductase derived from *Saccharomyces cerevisiae* or a functional variant thereof. In some embodiments, the introduced exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a truncated 3-hydroxy-3-methylglutaryl-CoA reductase. The truncated 3-hydroxy-3-methylglutaryl-CoA reductase may be referred to as tHMG1 and is a functional variant. In this study, truncated 3-hydroxy-3-methylglutaryl-CoA reductase can possess stronger catalytic activity than untruncated 3-hydroxy-3-methylglutaryl-CoA reductase. In some embodiments, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase can be a 3-hydroxy-3-methylglutaryl-CoA reductase with 528 amino acids removed from its N-terminus. In some embodiments, the introduced exogenous 3-hydroxy-3-methylglutaryl-CoA reductase comprises the amino acid sequence shown in SEQ ID NO. 2, preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is encoded by the nucleotide sequence of SEQ ID NO. 228.

[0127] In some embodiments, at least one gene modification enhancing the activity of mevalonate kinase includes: introducing an exogenous mevalonate kinase gene, increasing the copy number of the mevalonate kinase gene, and / or replacing the natural promoter of endogenous mevalonate kinase with a promoter that has a higher expression level. In some embodiments, the introduced exogenous mevalonate kinase gene encodes mevalonate kinase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous mevalonate kinase comprises the amino acid sequence shown in SEQ ID NO. 5, preferably, the exogenous mevalonate kinase is encoded by the nucleotide sequence of SEQ ID NO. 231.

[0128] In some embodiments, at least one gene modification enhancing the activity of mevalonate-5-phosphokinase includes: introducing an exogenous mevalonate-5-phosphokinase gene, increasing the copy number of the mevalonate-5-phosphokinase gene, and / or replacing the natural promoter of endogenous mevalonate-5-phosphokinase with a promoter having a higher expression level. In some embodiments, the introduced exogenous mevalonate-5-phosphokinase gene encodes mevalonate-5-phosphokinase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous mevalonate-5-phosphokinase gene comprises the amino acid sequence shown in SEQ ID NO. 6, preferably, the exogenous mevalonate-5-phosphokinase is encoded by the nucleotide sequence of SEQ ID NO. 232.

[0129] In some embodiments, at least one gene modification enhancing the activity of mevalonate-5-pyrophosphate decarboxylase includes: introducing an exogenous mevalonate-5-pyrophosphate decarboxylase gene, increasing the copy number of the mevalonate-5-pyrophosphate decarboxylase gene, and / or replacing the natural promoter of the endogenous mevalonate-5-pyrophosphate decarboxylase with a promoter having a higher expression level. In some embodiments, the introduced exogenous mevalonate-5-pyrophosphate decarboxylase gene encodes mevalonate-5-pyrophosphate decarboxylase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous mevalonate-5-pyrophosphate decarboxylase gene comprises the amino acid sequence shown in SEQ ID NO. 7, preferably, the exogenous mevalonate-5-pyrophosphate decarboxylase is encoded by the nucleotide sequence of SEQ ID NO. 233.

[0130] In some embodiments, at least one gene modification enhancing the activity of pentyrene pyrophosphate isomerase includes: introducing an exogenous pentyrene pyrophosphate isomerase gene, increasing the copy number of the pentyrene pyrophosphate isomerase gene, and / or replacing the natural promoter of the endogenous pentyrene pyrophosphate isomerase with a promoter having a higher expression level. In some embodiments, the introduced exogenous pentyrene pyrophosphate isomerase gene encodes a pentyrene pyrophosphate isomerase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous pentyrene pyrophosphate isomerase gene comprises the amino acid sequence shown in SEQ ID NO. 8, preferably, the exogenous pentyrene pyrophosphate isomerase is encoded by the nucleotide sequence of SEQ ID NO. 234.

[0131] In some embodiments, at least one gene modification enhancing the activity of farnesyl pyrophosphate synthase includes: introducing an exogenous farnesyl pyrophosphate synthase gene, increasing the copy number of the farnesyl pyrophosphate synthase gene, and / or replacing the natural promoter of endogenous farnesyl pyrophosphate synthase with a promoter having a higher expression level. In some embodiments, the introduced exogenous farnesyl pyrophosphate synthase gene encodes farnesyl pyrophosphate synthase derived from Saccharomyces cerevisiae or a functional variant thereof. In some embodiments, the introduced exogenous farnesyl pyrophosphate synthase gene comprises the amino acid sequence shown in SEQ ID NO. 1, preferably, the exogenous farnesyl pyrophosphate synthase is encoded by the nucleotide sequence of SEQ ID NO. 227.

[0132] Methods for genetically modifying engineered bacteria are well known to those skilled in the art. Genetic modification techniques can be used to introduce free exogenous nucleic acid sequences (e.g., in the form of plasmids) into engineered bacteria, or to insert exogenous nucleic acid sequences or delete endogenous nucleic acid sequences into the genome of engineered bacteria, or to replace a segment of endogenous nucleic acid sequence in the genome of engineered bacteria with a segment of exogenous nucleic acid sequence, so as to change the genes of engineered bacteria and thus change their phenotype.

[0133] Expressing exogenous proteins (such as one or more enzymes mentioned above) in engineered bacteria can include introducing exogenous genes into the engineered bacteria to express the protein encoded by those genes. This can be achieved, for example, by introducing a nucleic acid sequence (e.g., a vector) containing the exogenous gene into the engineered bacteria. The nucleic acid sequence containing the exogenous gene can be linear or circular, and can be single-stranded or double-stranded. The vector can be a self-replicating vector. The vector can be a free vector or an integrative vector. The vector can be, for example, a plasmid vector, a phage vector, a bacterial artificial chromosome, a transposon-based vector, or a CRISPR / Cas-based vector. The vector can also be a suicide vector, such as a suicide plasmid vector.

[0134] The expression vector may contain a foreign gene expression cassette, which may include a foreign gene and a regulatory sequence operatively linked thereto, the regulatory sequence guiding the expression of the foreign gene in suitable engineered bacteria. The regulatory sequence may include, but is not limited to, promoters, enhancers, terminators, and other expression control elements. The promoter may be a constitutive promoter to enable sustained expression of the foreign gene; or an inducible promoter to induce expression of the foreign gene upon the addition of an inducer. The vector may also contain one or more selectable marker genes that allow convenient selection of transformed, transfected, or transduced cells, such marker genes as genes providing resistance to antibiotics, heavy metals, and / or negative selection marker genes (e.g., ...). bag (B gene, etc.)

[0135] Suitable vectors can be selected based on different purposes (e.g., autonomous replication within engineered bacteria or integration into the genome of engineered bacteria) and / or different engineered bacteria. Vectors and regulatory sequences suitable for introducing exogenous proteins into different engineered bacteria are well known to those skilled in the art. For example, for *E. coli*, vectors that can be used include, but are not limited to, pJC1, pET22b(+), pBR322, pBR325, pUC57, pUC118, pUC119, pUC18, pUC19, pBluescript, or plasmids based on them; for *Corynebacterium glutamicum*, vectors that can be used include, but are not limited to, pBL1, pEKEx1, p... For Bacillus subtilis, plasmid vectors such as EKEx2, pXMJ19, pJC1, pHM1519, pVWEx1, pZ8-1, pECTAC-K99, pECTAC-XK99E, pECTAC-XC99E, pECTAC-XT99A, pNG2, and pAPE12, or plasmid vectors based thereon, may be used; for Bacillus subtilis, vectors that can be used include, but are not limited to, pHT43, pUB110, pE194, pUCX05-bgaB, and pWB980. pHP13, pBE2, pHP13, pHP13-43, pHT01, pHT304, pMK3, pHCMC05, pMA5, pHY300PLK, or pMUTIN4; for yeast, the vectors that can be used include, but are not limited to, pGal1, pGal7, pGal10, pGal2, pPIC9, pPIC9k, pHIL-S1, pPICza, pYAM75P6, pHIL-D2, pA0815, and pPIC3. K, pPICZ, pHWO10, pGAPZ, pGAPZa, pYES2, pYES2 / NT, pYES2 / CT, pYES3, pYES6, pYCplac22-GFP, pAUR123, pRS303TEF, pRS304, pRS305, pRS306, pY13TEF, pY14TEF, pY15TEF, pY16TEF, pSH47, pLacZi, pHIS2, or pGAD42. Suitable promoters for use in bacteria include, but are not limited to, inducible promoters such as Ptac, Plac, and Ptrc, or constitutive promoters such as Psod, PcspB, Ptuf, and PgapA. In some embodiments, promoters pGal1, pGal7, pGal10, and / or pGal2 are used to express exogenous genes in engineered bacteria.

[0136] The vector may also contain one or more selectable marker genes that allow for convenient selection of transformed, transfected, or transduced cells, such as genes that provide resistance to antibiotics or heavy metals.

[0137] The foreign gene, after being introduced into the engineered bacteria, can exist in and be expressed from a free vector (e.g., a free plasmid), for example, by introducing an expression vector (e.g., a plasmid vector) containing a foreign gene expression cassette into the engineered bacteria. The foreign gene can also be integrated into the genome of the engineered bacteria for expression, for example, by introducing an integrative plasmid vector containing the foreign gene (the plasmid may, for example, contain homologous arms for integration into the engineered bacteria genome via homologous recombination), a phage vector, a CRISPR / Cas system, or a transposon system (e.g., the Piggybac or Sleeping Beauty system). The foreign gene can be randomly integrated into the engineered bacteria, or it can be targeted to a suitable site in the engineered bacteria (e.g., via homologous recombination or a CRISPR / Cas system). The expression cassette containing the foreign gene can be randomly or targeted into the genome of the engineered bacteria, or the foreign gene can be targeted to a suitable site in the engineered bacteria to express the foreign gene using endogenous regulatory sequences within the engineered bacteria. In some implementations, the exogenous gene can be targeted and integrated into any one or more of the lpp1, dpp1, ho, gal80, gal2, dit1, and gal2 sites of the recombinant yeast engineer (e.g., recombinant Saccharomyces cerevisiae).

[0138] In some implementations, an expression cassette containing a foreign gene can be targeted and integrated into the genome of an engineered bacterium using a CRISPR / Cas system, or the foreign gene can be targeted and integrated into a suitable site within the engineered bacterium to express the foreign gene using endogenous regulatory sequences within the engineered bacterium. A CRISPR / Cas system typically comprises at least two components: a Cas9 protein, an enzyme capable of cleaving double-stranded DNA at a specific DNA sequence; and guide RNA (gRNA), which guides the Cas9 protein to a specific gene site for cleavage. Those skilled in the art are familiar with how to introduce foreign genes or knock out endogenous genes using a CRISPR / Cas9 system, for example, by constructing plasmids containing both the Cas9 protein and gRNA coding sequences. Donor plasmids are commonly used in CRISPR / Cas system-mediated gene editing technologies; donor plasmids may, for example, contain the complete expression cassette of a foreign gene (e.g., one or two or more foreign genes) and upstream and downstream homologous arms at the integration site. Donor plasmids may also contain regulatory sequences such as promoters and terminators that are operatively linked to foreign genes, as well as resistance genes and / or signal sequences.

[0139] Methods for expressing multiple foreign genes in engineered bacteria are well known to those skilled in the art. For example, a separate transcription unit can be constructed for each foreign gene, each with its own promoter and capable of being transcribed into its own mRNA. Appropriate promoters can be selected for different genes as needed. Alternatively, two or more (e.g., two, three, or four) foreign genes can be included in the same transcription unit, sharing the same promoter. Each of the multiple foreign genes in the same transcription unit may have a ribosome binding site (RBS) to facilitate individual translation of each gene, with multiple genes transcribed and translated in a polycistronic manner. Multiple transcription units can be contained in different vectors or in the same vector. These transcription units can be introduced into engineered bacteria using the methods described above.

[0140] Suitable methods for introducing exogenous nucleic acid sequences (e.g., vectors) into engineered bacteria are known to those skilled in the art, including but not limited to calcium phosphate transfection, protoplast fusion, electroporation, liposomes, lipid nanoparticles, microinjection, naked DNA or RNA (e.g., mRNA) transfection, plasmid vector transformation, phage vector transduction, etc.

[0141] In some implementations, based on the aforementioned genetic modification, the recombinant engineered bacteria can be further genetically modified to eliminate or weaken the role of one or more endogenous enzymes involved in competitive metabolic pathways.

[0142] Gene modifications that eliminate or attenuate endogenous enzymes include gene modifications that partially or completely inactivate the endogenous gene encoding the enzyme, including but not limited to: knocking out or knocking down the endogenous gene, for example, deleting all or part of the sequence of the endogenous gene, mutating the endogenous gene, or inserting a foreign sequence into the endogenous gene, so as to cause the enzyme activity encoded by the endogenous gene to be lost or attenuated. Gene modifications that partially or completely inactivate the endogenous gene encoding the enzyme may also include: knocking out or knocking down the natural regulatory sequence (e.g., the natural promoter) of the endogenous gene, for example, deleting all or part of the sequence of the regulatory sequence, mutating the regulatory sequence, or inserting a foreign sequence into the regulatory sequence, so as to partially or completely inactivate the regulatory sequence, thereby eliminating or reducing the expression of the endogenous gene.

[0143] By knocking out or downregulating endogenous genes, the proteins encoded by these endogenous genes in the engineered bacteria can be rendered nonfunctional or their activity reduced. This can be achieved, for example, by deleting all or part of the endogenous gene sequence, mutating the endogenous gene, or inserting an exogenous sequence into the endogenous gene. These gene modification techniques can also be used to alter the regulatory sequences (e.g., promoters) of endogenous genes to reduce their expression. Those skilled in the art can select appropriate methods to genetically modify engineered bacteria based on the engineered bacteria and the exogenous or endogenous nucleic acid sequences used (A Laboratory Manual (2nd Ed.), Vols. 1-3, Cold Spring Harbor Laboratory (1989) and Ausubel et al, eds., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York (1997)).

[0144] The engineered bacteria of this invention can be microbial cells, including bacteria, yeast cells, archaea, fungal cells, etc. The microorganisms can be GRAS (Generally Recognized As Safe) microorganisms. The bacteria can be, for example, Gram-negative or Gram-positive bacteria. The bacteria can be, for example, Escherichia bacteria, such as *Escherichia coli*. Escherichia coli Corynebacterium bacteria, such as Corynebacterium glutamicum (…); Corynebacterium glutamicum Corynebacterium pingeri () Corynebacterium pekinense ), Corynebacterium obliterans ( Corynebacterium crenatum ), Corynebacterium thermophilum ( Corynebacterium thermoaminogenes ), Corynebacterium ammoniagenic ( Corynebacterium aminogenes ) etc.; Bacillus bacteria, such as Bacillus subtilis ( Bacillus subtilis ), Bacillus licheniformis ( Bacillus licheniformis ), Bacillus coagulans ( Bacillus coagulans ), Bacillus cereus ( Bacillus cereus ), thermophilic steatobacterium ( Bacillus stearothermophilus ), Bacillus megaterium ( Bacillus megaterium ) etc.; Lactobacillus bacteria, such as Lactobacillus acidophilus ( Lactobacillus acidophilus ), Lactobacillus casei ( Lactobacillus casei Lactobacillus delbrueckii (), Lactobacillus delbrueckii Lactobacillus plantarum ( Lactococcus lactisBifidobacterium bacteria; Streptococcus bacteria; Lactococcus bacteria; Streptmyces bacteria; Pseudomonas bacteria, such as Pseudomonas aeruginosa. Pseudomonas aeruginosa Clostridium bacteria; Brevibacillus bacteria; Enterococcus bacteria; Pediococcus bacteria; Leuconostoc bacteria, etc. Yeast cells can be, for example, yeast cells of the genera *Saccharomyces*, *Saccharomycopsis*, *Pichia*, *Hansenula*, *Kluyveromyces*, *Yarrowia*, *Hyphozyma*, *Cryptococcus*, *Rhodotorula*, *Phaffia*, *Schizosaccharomyces*, or *Candida*, such as *Saccharomyces cerevisiae*. Saccharomyces cerevisiae ), Yarrowia lipolytica ( Yarrowia lipolytica ), Candida utilis ( Useful Candida ), or Pichia pastoris ( Peach shepherd ), Phaffia colomata ( Komagataellaphaffii) , Saccharomyces cerevisiae ( Schizosaccharomyces pombe ), Candida albicans ( Candida albicans ), Candida utilis ( White useful ), Yarrowia lipolytica ( Yarrowia lipolytica ), Hansenula polymorpha ( Hansenula polymorpha Pichia pastoris (Canada) Pichia canadensis ), Max Kluyveromycin ( Kluyveromyces marxianus Kluyveromycin (lactic acid yeast) Kluyveromyces lactis ), Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus albidus ) or Red Pavlova yeast ( Phaffia rhodozyma ).

[0145] In some embodiments, the engineered bacteria are bacteria or yeast; preferably, the bacteria are selected from Escherichia, Corynebacterium, or Bacillus, and the yeast is selected from Saccharomyces, Pichia, Hansenula, Kluyveromyces, Phaffia, Schizosaccharomyces, Candida, Yarrowia, Hyphozyma, or Cryptococcus; more preferably, the bacteria are selected from Escherichia coli (…). Escherichia coli ), Corynebacterium glutamicum ( Corynebacterium glutamicum ), or Bacillus subtilis ( Bacillus subtilis The yeast is selected from brewer's yeast (Saccharomyces cerevisiae). Saccharomyces yeast Pichia pastoris () Shepherd's pie ), Phaffia colomata ( Komagataellaphaffii) , Saccharomyces cerevisiae ( Schizosaccharomyces pombe ), Candida albicans ( Candida albicans ), Candida utilis ( Useful Candida ), Yarrowia lipolytica ( Yarrow lipolytic ), Hansenula polymorpha ( Hansenula polymorpha Pichia pastoris (Canada) Pichia canadensis ), Max Kluyveromycin ( Kluyveromycesmarxianus Kluyveromycin (lactic acid yeast) Kluyveromyces lactis ), Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus albidus ) or Red Pavlova yeast ( Phaffia rhodozyma More preferably, the brewing yeast is yeast strain CEN.PK2-1C.

[0146] In the method of the present invention, when the first component is contacted with the second component to produce salidomide and / or salidomide lactone, the content of lysalidomide may be from about 1 g / L to about 50 g / L, for example from about 5 g / L to about 20 g / L, from about 6 g / L to about 20 g / L, from about 7 g / L to about 20 g / L, from about 8 g / L to about 20 g / L, for example from about 8 g / L to about 10 g / L, for example from about 8.9 g / L. In some embodiments, when the second component is a microorganism capable of converting lysine-2-diol into styraxine and / or styraxine lactone, the first component may be added to the culture medium of the microorganism such that the lysine-2-diol content in the culture medium is about 1 g / L to about 50 g / L, for example about 5 g / L to about 20 g / L, about 6 g / L to about 20 g / L, about 7 g / L to about 20 g / L, about 8 g / L to about 20 g / L, for example about 8 g / L to about 10 g / L, for example about 8.9 g / L.

[0147] In some embodiments of the present invention, when the second component is a microorganism capable of converting lysine-2-diol into perillaldehyde and / or perillaldehyde lactone, the microorganism can be cultured in any suitable culture medium to produce perillaldehyde and / or perillaldehyde lactone. Such culture media are well known to those skilled in the art, including but not limited to YM medium, LPD medium, LG medium, etc. The culture medium may contain at least one carbon source, which may be selected from, but is not limited to, glucose, fructose, sucrose, acetic acid, glycerol, maltose, lactic acid, and / or succinic acid. In some embodiments, the carbon source concentration in the culture medium is from 10 g / L to 50 g / L, for example, 20 g / L to 50 g / L, 30 g / L to 50 g / L, 10 g / L to 40 g / L, 20 g / L to 40 g / L, or 30 g / L to 40 g / L. In some embodiments, the culture time of the microorganisms capable of converting lysine-3-diol into styraxine and / or styraxine lactone can be from about 12 hours to about 96 hours, for example from about 20 hours to about 72 hours, from about 24 hours to about 60 hours, or from about 36 hours to about 48 hours.

[0148] Mixed culture: In some embodiments, by co-culturing (1) a microorganism capable of producing lysine-1-diol (first microorganism) and (2) an enzyme or microorganism capable of converting lysine-1-diol into perillylene glycol and / or perillylene lactone (second microorganism), this invention uses lysine-1-diol as a raw material to produce perillylene lactone and / or perillylene glycol. This co-culturing method can efficiently and conveniently provide the raw material lysine-1-diol without the need for complex procedures to separate and purify lysine-1-diol produced by chemical or biosynthetic methods. In addition to the relatively cumbersome and difficult preparation and acquisition of lysine-1-diol, its production or purchase cost is also very high. In contrast, co-culturing can utilize very inexpensive glucose as a substrate to obtain the raw material lysine-1-diol through microbial fermentation. Furthermore, this method can obtain different proportions of perillylene glycol and perillylene lactone by controlling the cell density of the first and second microorganisms during co-culturing, and can also obtain single perillylene glycol or perillylene lactone. The co-culture of the two microorganisms provides a more efficient method for the production of perillaldehyde and / or perillyl glycol than existing technologies. The terms "mixed culture," "co-culture," and "co-culture" are used interchangeably herein and refer to the co-culture of two or more cell types (e.g., the first and second microorganisms of this invention) in the same environment, such as in the same culture medium, under the same conditions of temperature, carbon dioxide concentration, pH, dissolved oxygen concentration, etc. Typically, the two cell types in a co-culture are in direct physical contact.

[0149] Two or more cell types (e.g., the first and second microorganisms of the present invention) can be mixed and cultured in any suitable manner. In some embodiments, the two or more cell types can be inoculated into the same culture medium for co-culture. In some embodiments, each of the two or more cell types can be first fermented to obtain a cell culture medium (e.g., cultured to a certain cell density), and then mixed and cultured. In some embodiments, one or more (but not all) of the two or more cell types can be first fermented to obtain a cell culture medium (e.g., cultured to a certain cell density) and added to the co-culture medium, while other cell types are inoculated into the co-culture medium. They are then mixed and cultured. In some embodiments, each of the two or more cell types can be cultured for about 12 hours to about 96 hours, for example, about 20 hours to about 72 hours, about 24 hours to about 60 hours, or about 36 hours to about 48 hours before being mixed with other cells. In some embodiments, any one of the two or more cell types is cultured to an OD value (e.g., OD600) of about 1.0 to about 50, such as about 5.0 to about 30, about 10 to about 30, about 10 to about 20, or about 20 to about 30 before being mixed with other cells.

[0150] In some embodiments, the second microorganism is induced to culture before being mixed with the first microorganism, for example, by using lysine diol as an inducer. In some embodiments, the induction culture is carried out for about 12 hours to about 96 hours, for example, about 20 hours to about 72 hours, about 24 hours to about 60 hours, or about 36 hours to about 48 hours. In some embodiments, the concentration of the inducer during the induction culture is about 0.01 g / L to about 1 g / L, for example, about 0.01 g / L to about 0.5 g / L, about 0.05 g / L to about 0.5 g / L, about 0.05 g / L to about 0.4 g / L, about 0.05 g / L to about 0.3 g / L, about 0.05 g / L to about 0.2 g / L, about 0.05 g / L to about 0.1 g / L, or about 0.1 g / L to about 0.2 g / L. Induction culture enables the microorganisms to produce better results in subsequent co-culture, for example, it enables the second microorganism to have a stronger absorption or utilization effect on lysine diol. When the second microorganism is induced and then co-cultured with the first microorganism, the yield of perillaldehyde and / or perillyl lactone is higher than that of the second microorganism without induction culture co-cultured with the first microorganism.

[0151] Two or more cell types (e.g., the first and second microorganisms of the present invention) can be mixed in any suitable ratio and then subjected to co-culture. In some embodiments, the cell density ratio of the first and second microorganisms when mixed is about 20:1 to about 1:20. In some specific implementation schemes, the cell density ratio of the first microorganism and the second microorganism when mixed is about 20:1, about 19:1, about 18:1, about 17:1, about 16:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, or about 1:20. In some implementations, cell density can be measured as an OD600 value or the number of cells per unit volume.

[0152] In some embodiments, the cell density ratio of the first microorganism and the second microorganism when mixed is 10.5:1, 8.5:1, 4.8:1, 3.9:1, 2:1, 1.9:1, 1.55:1, 1:1, 0.96:1, 0.78:1, 1:2, 1:2.1, 1:2.58, 1:5.2, 1:6.45, 1:11.5 or 1:14.

[0153] In some implementations, when the second microorganism is a fungus with accession number ATCC 20624, the cell density ratio of the first microorganism and the second microorganism when mixed is 8.5:1, 3.9:1, 2:1, 1.55:1, 1:1, 0.78:1, 1:2, 1:2.58 or 1:6.45.

[0154] In some implementations, when the second microorganism is a fungus with accession number ATCC 20918, the cell density ratio of the first microorganism and the second microorganism when mixed is 10.5:1, 4.8:1, 2:1, 1.9:1, 1:1, 0.96:1, 1:2, 1:2.1, 1:5.2 or 1:11.5.

[0155] In some embodiments, the culture temperature of the co-culture is above about 15°C; preferably, the culture temperature of the co-culture is from about 15°C to about 40°C, more preferably, the culture temperature of the co-culture is from about 20°C to about 35°C, and more preferably, the culture temperature of the co-culture is from about 25°C to about 30°C. In some embodiments, when the second microorganism is a fungus with accession number ATCC 20624, the culture temperature of the co-culture is about 25°C, at which temperature a higher yield of perillaldehyde and / or perillyl lactone can be obtained. In some embodiments, when the second microorganism is a fungus with accession number ATCC 20918, the culture temperature of the co-culture is about 30°C, at which temperature a higher yield of perillaldehyde and / or perillyl lactone can be obtained. The analysis, detection, and / or quantification of perillyl lactone and / or perillaldehyde can be performed using techniques well known in the art, such as gas chromatography.

[0156] The first microorganism and / or the second microorganism can be co-cultured using any suitable culture medium, including but not limited to media suitable for bacteria, fungi, or yeast, such as YM medium, LPD medium, LG medium, etc. Those skilled in the art are familiar with culture media suitable for culturing different microorganisms; therefore, a medium suitable for the growth of both the first and second microorganisms can be selected for co-culture. The culture medium may contain at least one carbon source, which may be selected from, but is not limited to, glucose, fructose, sucrose, acetic acid, glycerol, maltose, lactic acid, and / or succinic acid. In some embodiments, the carbon source concentration in the culture medium is from 10 g / L to 50 g / L, for example, 20 g / L to 50 g / L, 30 g / L to 50 g / L, 10 g / L to 40 g / L, 20 g / L to 40 g / L, or 30 g / L to 40 g / L. In some embodiments, the LG culture medium of the present invention comprises the following components: ammonium sulfate, potassium dihydrogen phosphate; magnesium sulfate heptahydrate; biotin; calcium pantothenate; nicotinic acid; inositol; thiamine hydrochloride; pyridoxal hydrochloride; para-aminobenzoic acid; zinc sulfate heptahydrate; manganese chloride tetrahydrate; copper sulfate pentahydrate; cobalt chloride hexahydrate; sodium molybdate dihydrate; ferrous sulfate heptahydrate; calcium chloride dihydrate; 0.1168 g / L; succinic acid; glucose; yeast extract, adjusted to pH 6.0. In some embodiments, the LG culture medium of the present invention comprises the following components: ammonium sulfate 15 g / L, potassium dihydrogen phosphate 8 g / L; magnesium sulfate heptahydrate 6.15 g / L; biotin 0.0006 g / L; calcium pantothenate 0.012 g / L; nicotinic acid 0.012 g / L; inositol 0.3 g / L; thiamine hydrochloride 0.012 g / L; pyridoxal hydrochloride 0.012 g / L; p-aminobenzoic acid 0.0024 g / L; zinc sulfate heptahydrate 0.0575 g / L; manganese chloride tetrahydrate 0.0032 g / L; copper sulfate pentahydrate 0.0032 g / L; cobalt chloride hexahydrate 0.0047 g / L; sodium molybdate dihydrate 0.0048 g / L; ferrous sulfate heptahydrate 0.028 g / L; calcium chloride dihydrate 0.029 g / L; ethylenediaminetetraacetic acid 0.1168 g / L. g / L; succinic acid 5.9 g / L; glucose 40 g / L; 5 g / L yeast extract, adjust pH to 6.0.

[0157] In some implementations, during the co-culture process, a carbon source, such as glucose, can be supplemented at appropriate times. The concentration of the supplemented carbon source can be, for example, a final concentration of 10 g / L to 50 g / L, such as 20 g / L to 50 g / L, 30 g / L to 50 g / L, 10 g / L to 40 g / L, 20 g / L to 40 g / L, or 30 g / L to 40 g / L.

[0158] The mixed culture mode can be batch fermentation, where a closed culture system and specific culture medium are used at the beginning of fermentation, and specific temperature, pressure, aeration, and other environmental conditions are used to optimize growth. No nutrients are added during cell culture, and no fermentation broth is released. The culture of engineered bacteria can also be fed-batch fermentation, where nutrients are added intermittently or continuously during fermentation, but no fermentation broth is released. The culture of engineered bacteria can also be continuous fermentation, where nutrients are continuously added and fermentation broth is continuously released during fermentation, thereby maintaining a constant volume of culture medium in the fermentation system. The mixed culture can also be a combination of two or three of the above fermentation methods.

[0159] In some implementations, the co-culture can be carried out for an appropriate time, such as about 12 hours to about 96 hours, for example about 20 hours to about 72 hours, about 24 hours to about 60 hours, or about 36 hours to about 48 hours.

[0160] In some embodiments, the production method of the present invention (including but not limited to the mixed culture method) includes isolating, purifying or recovering the synthesized perillaldehyde and / or perillyl lactone from the culture medium and / or from the microorganisms.

[0161] For example, the product can be separated, purified, or recovered from the culture medium and / or the supernatant of the cell lysate. Cell lysis can be performed by chemical or physical methods known in the art. The terms “separation,” “purification,” and “recovery” refer to the separation of the product from other components in the engineered bacterial culture, which may include, for example, the removal of impurities and unwanted byproducts, such as cells, ions, salts, and / or substances other than the desired compound of formula (I), such as farnesol, geraniol, and lysine diol.

[0162] Separation, purification, or recovery can be performed using techniques known to those skilled in the art. For example, the product can be purified from the culture medium using methods known to those skilled in the art, such as column chromatography with activated carbon, elution with a concentration gradient of ethanol, or size exclusion chromatography or ion exchange chromatography. Purity can be assessed by any known method, such as thin-layer chromatography or other electrophoretic or chromatographic techniques commonly known in the art.

[0163] The perillaldehyde and / or perillyl lactone produced using the method of this invention can be used to further produce derivatives thereof, for example, by converting the compound of formula (I) into its derivatives through chemical synthesis, biocatalysis (e.g., by enzyme-catalyzed reactions), or a combination of both. The chemical synthesis includes, but is not limited to, oxidation, reduction, alkylation, acylation, and / or rearrangement. The biocatalysis involves catalyzing the perillaldehyde and / or perillyl lactone by contacting an enzyme, such as, but not limited to, oxidoreductases, monooxygenases, dioxygenases, and transferases. The derivatives may be, for example, hydrocarbons, diols, triols, acetals, ketals, aldehydes, acids, ethers, amides, ketones, epoxides, acetates, glycosides, and / or esters.

[0164] When further producing derivatives of perillaldehyde and / or perillyl lactone using the method of the present invention, the perillaldehyde and / or perillyl lactone may be isolated from the reaction mixture of the method or from the culture of the recombinant engineered bacteria, or may not be isolated. The reaction mixture or the recombinant engineered bacteria may be used directly as a raw material to provide perillaldehyde and / or perillyl lactone for the derivatives.

[0165] When the second microorganism is a fungus with accession number ATCC 20624, the co-culture can produce perillaldehyde at concentrations of about 6 mg / L or more, for example, about 6 mg / L or more, 8 mg / L or more, 10 mg / L or more, 12 mg / L or more, 24 mg / L or more, 30 mg / L or more, 35 mg / L or more, 40 mg / L or more, 45 mg / L or more, 90 mg / L or more, 150 mg / L or more, or 160 mg / L or more.

[0166] When the second microorganism is a fungus with accession number ATCC 20624, the co-culture can produce styracil lactone at a concentration of about 5 mg / L or more, for example, about 5 mg / L or more, 8 mg / L or more, 10 mg / L or more, 12 mg / L or more, 24 mg / L or more, 30 mg / L or more, or 35 mg / L or more.

[0167] In some implementations, when the second microorganism is a fungus with accession number ATCC 20624, the co-culture temperature is about 30°C, and the cell density ratio of the second microorganism to the first microorganism is 1:1.5, the yield of perillaldehyde can reach 164.7 mg / L, which is much higher than the yield of co-culture at about 25°C, and trace amounts of perillaldehyde can be detected.

[0168] When the second microorganism is a fungus with accession number ATCC 20918, the co-culture can produce perillaldehyde at a concentration of about 6 mg / L, for example, about 6 mg / L or more, 8 mg / L or more, 10 mg / L or more, 12 mg / L or more, 24 mg / L or more, 30 mg / L or more, 35 mg / L or more, 40 mg / L or more, 45 mg / L or more, 50 mg / L or more, 60 mg / L or more, or 70 mg / L or more.

[0169] When the second microorganism is a fungus with accession number ATCC 20918, the co-culture can produce styracil lactone at a concentration of about 5 mg / L, for example, about 5 mg / L or more, 8 mg / L or more, 10 mg / L or more, 12 mg / L or more, 24 mg / L or more, 30 mg / L or more, or 35 mg / L or more.

[0170] In some implementations, when the second microorganism is a fungus with accession number ATCC 20918, the co-culture temperature is approximately 25°C, and the initial addition amount of the fungus with accession number ATCC 20918 is [OD value missing]. 600 The highest yield of perilla lactone was observed at a ratio of 1:10.5, reaching 77.4 mg / L. Composition: Two or more cell types (e.g., the first and second microorganisms of the present invention) can be mixed or formulated into compositions for the production of perillaldehyde and / or perillyl lactone. In some embodiments, compositions comprising the first and second microorganisms can be co-cultured to produce perillaldehyde and / or perillyl lactone.

[0171] The ratio of the first microorganism to the second microorganism in the composition can be approximately 20:1 to approximately 1:20. In some specific implementation schemes, the ratio of the first microorganism to the second microorganism can be approximately 20:1, approximately 19:1, approximately 18:1, approximately 17:1, approximately 16:1, approximately 15:1, approximately 14:1, approximately 13:1, approximately 12:1, approximately 11:1, approximately 10:1, approximately 9:1, approximately 8:1, approximately 7:1, approximately 6:1, approximately 5:1, approximately 4:1, approximately 3:1, approximately 2:1, approximately 1:1, approximately 1:2, approximately 1:3, approximately 1:4, approximately 1:5, approximately 1:6, approximately 1:7, approximately 1:8, approximately 1:9, approximately 1:10, approximately 1:11, approximately 1:12, approximately 1:13, approximately 1:14, approximately 1:15, approximately 1:16, approximately 1:17, approximately 1:18, approximately 1:19, or approximately 1:20. In some implementations, the ratio of the first microorganism to the second microorganism can be measured by cell density or cell number, for example, by the ratio of OD600 values.

[0172] In some embodiments, the ratio of the first microorganism to the second microorganism in the composition may be 10.5:1, 8.5:1, 4.8:1, 3.9:1, 2:1, 1.9:1, 1.55:1, 1:1, 0.96:1, 0.78:1, 1:2, 1:2.1, 1:2.58, 1:5.2, 1:6.45, 1:11.5, or 1:14.

[0173] In some embodiments, when the second microorganism is a fungus with accession number ATCC 20624, the ratio of the first microorganism to the second microorganism in the composition can be 8.5:1, 3.9:1, 2:1, 1.55:1, 1:1, 0.78:1, 1:2, 1:2.58, or 1:6.45.

[0174] In some embodiments, when the second microorganism is a fungus with accession number ATCC 20918, the ratio of the first microorganism to the second microorganism in the composition can be 10.5:1, 4.8:1, 2:1, 1.9:1, 1:1, 0.96:1, 1:2, or 1:2.1, 1:5.2, or 1:11.5.

[0175] In some embodiments, two or more cell types in the composition (e.g., the first and second microorganisms of the present invention) are cultured via fermentation (e.g., cultured to a certain cell density). The culture may be, for example, about 12 hours to about 96 hours, about 20 hours to about 72 hours, about 24 hours to about 60 hours, or about 36 hours to about 48 hours. In some embodiments, the two or more cell types in the composition (e.g., the first and second microorganisms of the present invention) are cultured to an OD value (e.g., OD600) of about 1.0 to about 50, for example, about 5.0 to about 30, about 10 to about 30, about 10 to about 20, or about 20 to about 30.

[0176] In some embodiments, the second microorganism in the composition is induced cultured, for example, using lysine diol as an inducer. In some embodiments, the induction culture is carried out for about 12 hours to about 96 hours, for example, about 20 hours to about 72 hours, about 24 hours to about 60 hours, or about 36 hours to about 48 hours. In some embodiments, the concentration of the inducer during the induction culture is about 0.01 g / L to about 1 g / L, for example, about 0.01 g / L to about 0.5 g / L, about 0.05 g / L to about 0.5 g / L, about 0.05 g / L to about 0.4 g / L, about 0.05 g / L to about 0.3 g / L, about 0.05 g / L to about 0.2 g / L, about 0.05 g / L to about 0.1 g / L, or about 0.1 g / L to about 0.2 g / L.

[0177] the term The term "gene" refers to the nucleotide sequence that encodes a gene product. The gene product can be a protein or ribonucleic acid.

[0178] The terms “nucleic acid,” “nucleic acid sequence,” or “polynucleotide” refer to a single- or double-stranded polymer of deoxynucleotide or ribonucleotide bases, including DNA or RNA, including linear or circular DNA or RNA.

[0179] The terms "peptide" and "protein" are used interchangeably, referring to polymers of amino acid residues. The enzyme in this invention is a protein capable of catalyzing a chemical reaction of its substrate.

[0180] The term "cell lysate" refers to substances obtained by disrupting the cell membrane and releasing the cell contents. Disruption of the cell membrane (or cell lysis) can be achieved by any suitable means known to those skilled in the art, such as physical and / or chemical methods.

[0181] The term "enzyme extract" refers to a fraction containing an enzyme or capable of exerting enzymatic activity, extracted from cells or cell cultures. Enzyme extracts can be crude enzyme extracts or purified enzymes, or lyophilized crude or purified enzymes. Enzyme extracts can be a mixture of multiple enzymes. In some embodiments, the enzyme extract is a total protein extract from microorganisms. In some embodiments, the enzyme extract can be a supernatant obtained from microbial lysis by standing or centrifuging cell lysates, and may contain very little or virtually no cell membrane components.

[0182] The term "recombinant" when used to refer to cells, nucleic acids, proteins, or vectors indicates that the cells, nucleic acids, proteins, or vectors have been modified by introducing heterologous nucleic acids or proteins or by altering native nucleic acids or proteins, or that the cells are derived from cells that have been modified in this way. Therefore, recombinant cells express genes that are not present in the natural (non-recombinant) form of the cell; or express natural genes that are otherwise abnormally, insufficiently, or not expressed at all.

[0183] The terms "genetic modification," "genetic alteration," "engineering," and "engineering" refer to the artificial manipulation of altering the sequence of a polypeptide or polynucleotide, or altering the gene sequence contained in an engineered bacterium, to include a sequence not naturally present in the polypeptide, polynucleotide, or engineered bacterium. When an engineered bacterium is "engineered" to exhibit a certain characteristic (e.g., containing a gene encoding a specific protein, expressing a specific protein, or overexpressing a specific gene), it means that the engineered bacterium did not possess that characteristic before modification but acquires it after modification. The term "synthetic pathway" refers to a series of enzyme-controlled and catalyzed reactions that result in the synthesis of a chemical substance, such as lysandrindiol (LOH), lysandrindiol pyrophosphate (LPP), or farnesyl pyrophosphate (FPP) as described herein. In some embodiments, the term "synthetic pathway" may include a series of reactions starting from a carbon source and catalyzed by an enzyme to synthesize a specific substance. The term "synthetic pathway gene" refers to the gene encoding an enzyme that catalyzes a series of reactions included in a synthetic pathway, and these genes may be expressed in engineered bacteria to catalyze the synthesis of a specific chemical substance.

[0184] The term "exogenous" is used in contrast to engineered bacteria, referring to substances or molecules that originate from or are produced outside of engineered bacteria. "Exogenous gene" or "exogenous enzyme" refers to nucleic acids that are not naturally occurring genes or enzymes in the cell, but are introduced into the cell artificially. The sequence of an exogenous gene or exogenous enzyme may be the same as or different from the naturally occurring endogenous sequence in the cell. Genes or enzymes that differ from the naturally occurring endogenous sequence in the cell are called heterologous genes or heterologous enzymes, and they may originate from strains or species that are the same as or different from engineered bacteria. In this article, unless otherwise stated, the use of "exogenous gene" or "exogenous enzyme" includes the case of "heterologous gene" or "heterologous enzyme."

[0185] The term "endogenous" refers to genes or proteins (e.g., wild-type genes or wild-type enzymes) or synthetic pathways that are naturally present in engineered bacteria.

[0186] The term "wild-type" refers to a naturally occurring organism or cell, or a nucleic acid sequence present in a naturally occurring organism or cell, or a protein expressed by a naturally occurring organism or cell.

[0187] The term "naturally occurring" refers to nucleic acid sequences, amino acid sequences, complexes, pathways, or cells that exist in nature without human intervention.

[0188] The term "separated" means that a component, material, or substance has been separated from the environment in which it was produced. "Separated" can refer to a component, material, or substance being separated from other components, materials, or substances normally associated with it in the environment in which it was produced, and resulting in an increase in purity due to this separation. The purity of a "separated" component, material, or substance can be at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by weight.

[0189] The term "derived from" when applied to a protein or gene sequence refers to a protein or gene sequence that is derived from a specific organism, meaning that the protein or gene sequence has the same structure or sequence as proteins or gene sequences naturally present in that organism, and is not limited to being directly isolated from that organism.

[0190] The term "mutant" refers to a polypeptide that has one or more amino acid insertions, deletions, and / or substitutions relative to the parent polypeptide. Substitution means replacing an amino acid occupying a position with a different amino acid; deletion means removing an amino acid occupying a position; insertion means adding one or more amino acids (e.g., 1-5) adjacent to an amino acid occupying a position. Mutants retain at least one activity of the parent polypeptide, but may have variations at the activity level; for example, a mutant may remain unchanged or improve upon at least one activity or property relative to the parent polypeptide.

[0191] The term "functional variant," when applied to polypeptides or proteins, refers to a polypeptide that shares at least 60%, 65%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with its parent polypeptide and has the same or substantially the same function as the parent polypeptide. A functional variant may also refer to a polypeptide that has the insertion, deletion, and / or substitution of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) compared to its parent polypeptide and has the same or substantially the same function as the parent polypeptide. Modifications or methods for creating functional variants of polypeptides or proteins are well known to those skilled in the art. Polypeptides known in the art that have the same or substantially the same function as their parent polypeptide are also included within the scope of the enzymes or their functional variants of the present invention.

[0192] The term "parent" refers to a polypeptide that has been modified to produce a mutant, and the parent can be a naturally occurring (wild-type) polypeptide or a mutant thereof.

[0193] The term “sequence identity” refers to the percentage of identical nucleotide or amino acid residues at corresponding positions in two or more sequences, when sequences are aligned to maximize sequence matching, i.e., gaps and insertions are taken into account. Sequence alignment and the calculation of sequence identity percentages can be performed using appropriate computer programs known in the art. These programs include, but are not limited to, BLAST, ALIGN, ClustalW, EMBOSS Needle, etc. For example, global sequence alignment can be performed based on the Needleman-Wunsch algorithm (Needleman, Saul B.; and Wunsch, Christian D. (1970), “A general method applicable to the search for similarities in the amino acidsequence of two proteins”, Journal of Molecular Biology 48(3): 443-53), which is available at http: / / www.ebi.ac.uk / Tools / psa / and can be performed using default parameters. Local alignment can be performed using BLAST (Basic Local Alignment Search Tool), first described in Altschul et al. (1990) J. Mol. Biol. 215; 403. Biol. 215; 403-410. The BLAST alignment tool is available from the website of the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / / ), and default parameters can be used for alignment, for example.

[0194] The term "vector" refers to a tool that allows or facilitates the transfer of nucleic acid fragments from one environment to another (such as engineered bacteria). Vectors allow the insertion of another nucleic acid fragment to achieve replication of the inserted fragment.

[0195] The term "expression vector" refers to a vector used to express a gene product, which typically includes one or more expression control sequences for controlling and regulating the transcription and / or translation of a gene sequence that can express the product.

[0196] The term "expression cassette" refers to a nucleotide sequence containing relevant nucleic acids that are controlled by and operatively linked to appropriate promoters or other regulatory elements to enable transcription of the relevant nucleic acids in engineered bacteria.

[0197] The term "transcription unit" refers to a nucleic acid sequence containing one or more genes to be transcribed. Genes within a transcription unit are operatively linked to each other in a manner such that all genes within the unit are under the transcriptional control of the same promoter and / or enhancer, thereby enabling the transcription of more than one protein or product.

[0198] The term "operable linking" refers to placing a regulatory sequence necessary for the expression of a coding sequence in the appropriate position within the DNA molecule relative to the coding sequence, thereby affecting the expression of the coding sequence.

[0199] The term "knockout" refers to the manipulation of genes to prevent cells or organisms from producing the functional products encoded by those genes.

[0200] The term "knockdown" refers to the process of manipulating genes to reduce the functional activity of functional products encoded by said genes produced by cells or organisms.

[0201] The term "eliminating or weakening the effect of an enzyme" means that the reaction catalyzed by the enzyme is eliminated or weakened, which can be specifically manifested as the absence of reaction products catalyzed by the enzyme or a reduction in the amount of products generated by the reaction catalyzed by the enzyme.

[0202] The term "enhancing enzyme action" refers to the enhancement of the reaction catalyzed by the enzyme, which can be specifically manifested as an increase in the amount of product generated by the reaction catalyzed by the enzyme.

[0203] The term "enzyme activity" refers to the ability of an enzyme to catalyze the conversion of a substrate into a product, which can be evaluated by the amount of product produced.

[0204] The term "overexpression" refers to the expression level of a gene product or peptide in engineered bacteria that is higher after genetic modification than before the modification. "Overexpression" can also refer to any detectable expression caused by introducing a specific gene product into the engineered bacteria if the bacteria did not contain the specific gene product before modification.

[0205] The term "natural promoter" refers to the promoter of a gene that is naturally present in engineered bacteria.

[0206] The term "homological protein" refers to a protein that has similar activity and / or similar structure to a protein of interest (e.g., a reference protein). In some embodiments, the sequence identity of the homologous protein to the reference protein is at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0207] The term "acetyl-CoA transferase," also known as "acetoacetyl-CoA thiolysis enzyme," refers to the enzyme that catalyzes the conversion of acetyl-coenzyme A (acetyl-CoA) to acetoacetyl-CoA. Acetyl-CoA transferase may be referred to as ERG10.

[0208] The term "3-methyl-3-hydroxyglutaryl-CoA synthase," also known as "hydroxymethylglutaryl-CoA synthase" or "3-hydroxy-3-methylglutary-CoA synthase (HMGS)," refers to the enzyme that catalyzes the synthesis of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) from acetoacetyl-CoA. 3-methyl-3-hydroxyglutaryl-CoA synthase may also be referred to as ERG13.

[0209] The term "3-hydroxy-3-methylglutaryl-CoA reductase," also known as "hydroxymethylglutaryl-CoA reductase (HMGR)," and referred to herein as "untruncated 3-hydroxy-3-methylglutaryl-CoA reductase," refers to the enzyme that catalyzes the formation of mevalonic acid (MVA) from 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA). 3-hydroxy-3-methylglutaryl-CoA reductase may be referred to as HMG1.

[0210] The term "mevalonate kinase," also known as "MK," refers to the enzyme that catalyzes the conversion of mevalonate (MVA) to mevalonate phosphate (MVAP). Mevalonate kinase may also be referred to as ERG12.

[0211] The term "mevalonate-5-phosphate kinase," also known as "phosphomevalonate kinase (PMK)," refers to the enzyme that catalyzes the conversion of mevalonate phosphate (MVAP) to mevalonate pyrophosphate (MVAP). Mevalonate-5-phosphate kinase can also be referred to as ERG8.

[0212] The term "5-pyrophosphate decarboxylase," also known as "pyrophosphate mevalonate decarboxylase (MDC)," refers to the enzyme that catalyzes the formation of isopentenyl pyrophosphate (IPP) from mevalonate pyrophosphate (MVAP). 5-pyrophosphate decarboxylase can also be referred to as ERG19.

[0213] The term "pentene pyrophosphate isomerase," also known as "isopentenyldiphosphate isomerase (IDI)," refers to the enzyme that catalyzes the formation of dimethylallyl diphosphate (DMAPP) from isopentenyl pyrophosphate (IPP), or vice versa. Pentene pyrophosphate isomerase can also be referred to as IDI1.

[0214] The term "farnesyl pyrophosphate synthase," also known as "farnesyl pyrophosphate synthase (FPS / ERG20)," refers to the enzyme that catalyzes the synthesis of farnesyl diphosphate (FPP) from IPP and DMAPP. Farnesyl pyrophosphate synthase can also be referred to as ERG20.

[0215] The term "geranylgeranyl pyrophosphate synthase," also known as "geranylgeranyl diphosphate synthase (GGPPS)," refers to the enzyme that catalyzes the conversion of farnesyl diphosphate (FPP) to geranylgeranyl diphosphate (GGPP). Geranylgeranyl pyrophosphate synthase can also be referred to as BTS1.

[0216] The term "lysandrin diol pyrophosphate synthase" refers to the enzyme that catalyzes the conversion of geranylgeranyl diphosphate (GGPP) to lysandrin diol pyrophosphate (LPP). Lysandrin diol pyrophosphate synthase can also be referred to as LPPS.

[0217] It should be understood that for enzymes with specific activities, the sequences of homologous proteins in different organisms (including different species or different strains of the same species) are readily available to those skilled in the art, for example, by searching public databases (e.g., GenBank).

[0218] Enzymes and their encoding genes with the same biological activity are known in the art to have different names, sometimes related to the microorganism from which the enzyme originates. The enzymes described herein are intended to cover enzymes with the defined functions, and unless otherwise stated, should cover enzymes with the defined functions from any organism (including microorganisms, animals, etc.).

[0219] In this invention, it should be understood that when an abbreviation is shown in parentheses after the name of an enzyme, the abbreviation should not be regarded as a limitation on a specific sequence or specific source of the enzyme, but is merely an example for reference and understanding.

[0220] Unless otherwise stated, nucleic acids are written from left to right in the 5' to 3' direction in this document, and amino acid sequences are written from left to right in the direction from the amino terminus to the carboxyl terminus.

[0221] The present invention is further described through the following embodiments, which should not be construed as limiting the present invention.

[0222] Unless otherwise specified, all reagents used in the following examples are commercially available products. Molecular biology experimental methods not specifically described in the examples were performed according to the specific methods listed in J. Sambrook, Molecular Cloning: A Laboratory Manual, Third Edition, or according to the kit and product instructions.

[0223] Example 1: Culture medium used for strain culture and product detection The seed culture medium used for fungi ATCC 20918 and ATCC 20624 was YM medium, with the following components: yeast extract 3 g / L, peptone 5 g / L, malt extract 3 g / L, glucose 10 g / L, and pH adjusted to 6.2. The YM seed culture medium was sterilized by filtration through a 0.22 μm filter. Adding 2% agar to the medium prepared solid plates.

[0224] The fermentation medium LGM components used in ATCC 20918 and ATCC 20624 are as follows: ammonium sulfate 15 g / L, potassium dihydrogen phosphate 8 g / L; magnesium sulfate heptahydrate 6.15 g / L; biotin 0.0006 g / L; calcium pantothenate 0.012 g / L; nicotinic acid 0.012 g / L; inositol 0.3 g / L; thiamine hydrochloride 0.012 g / L; pyridoxal hydrochloride 0.012 g / L; p-aminobenzoic acid 0.0024 g / L; zinc sulfate heptahydrate 0.0575 g / L; manganese chloride tetrahydrate 0.0032 g / L; copper sulfate pentahydrate 0.0032 g / L; cobalt chloride hexahydrate 0.0047 g / L; sodium molybdate dihydrate 0.0048 g / L; ferrous sulfate heptahydrate 0.028 g / L; calcium chloride dihydrate 0.029 g / L. The following were added: 0.1168 g / L EDTA; 5.9 g / L succinic acid; 40 g / L glucose; 5 g / L yeast extract. The pH was adjusted to 6.0. The fermentation medium LGM was sterilized by filtration through a 0.22 μm filter membrane.

[0225] The seed culture medium (YPD) used for the lysine-producing strain consisted of 10 g / L yeast extract, 20 g / L peptone, and 20 g / L glucose. Adding an additional 2% agar to the medium prepared solid plates.

[0226] The fermentation medium used for the lysine-producing strain was the LGM fermentation medium mentioned above.

[0227] The analysis and detection of substances such as Labdenediol, Sclareol, sclareolglycol, and Sclareolide were performed using an Agilent GC8890. Column: Agilent HP-5 (30 m, 0.32 mm, 0.25 μm, P / N 19091J-413-KEY); Carrier gas: High-purity nitrogen; Flow rate: 2.5 mL / min; Injection volume: 1 μL; Injector temperature: 280℃; Split ratio: 50:1; Detector: FID; Detector temperature: 300℃; Air flow rate: 300 mL / min; Hydrogen flow rate: 30 mL / min; Make-up gas flow rate: 25 mL / min; Temperature program: 150℃ held for 1.5 min, increased to 230℃ at a rate of 20℃ / min, held for 1 min, then increased to 300℃ at a rate of 60℃ / min, held for 2 min; Quantification was performed using the internal standard method.

[0228] Example 2: Production of Lysadione using Saccharomyces cerevisiae genetically engineered strain YC6 The method for constructing engineered strains that produce lysine-3-diol is to introduce a screened phosphorylase into Saccharomyces cerevisiae.

[0229] Starting with strains rich in the precursor farnesyl pyrophosphate (FPP), numerous Saccharomyces cerevisiae genetically engineered strains capable of producing lysenoside diol were constructed by simultaneously overexpressing the geraniol-geraniol pyrophosphate synthase gene BTS1, the lysenoside diol pyrophosphate ester synthase SsLPPS, and screened phosphorylases. Among these, the Saccharomyces cerevisiae genetically engineered strain YC6 produced the highest yield of lysenoside diol.

[0230] The method for constructing the recombinant Saccharomyces cerevisiae strain YC6 is as follows: Constructing a background yeast strain for producing lysine diol This invention uses the wild-type yeast strain CEN.PK2-1C (purchased from Euroscarf) as the starting strain for the production of lysenoside diol. CRISPR-Cas9-mediated genome editing is a commonly used synthetic biology tool in Saccharomyces cerevisiae. The CRISPR-Cas9 method was used to integrate key enzymes in the lysenoside diol biosynthesis pathway into the genome. One copy of the farnesyl pyrophosphate synthase gene ERG20 (amino acid sequence SEQ ID NO.1, nucleotide sequence SEQ ID NO.227) and one copy of a truncated 3-hydroxy-3-methylglutaryl-CoA reductase tHMG1 (amino acid sequence SEQ ID NO.2, nucleotide sequence SEQ ID NO.228) were inserted at the lpp1 site in the genome. The gRNA-N20 sequence used was ATGTAAAACTGACGTTCGAA (SEQ ID NO.454), and the corresponding gRNA-1 plasmid was constructed. One copy of acetyl-CoA transferase ERG10 (amino acid sequence SEQ ID NO.3, nucleotide sequence SEQ ID NO.229) and one copy of 3-methyl-3-hydroxyglutaryl-CoA synthase ERG13 (amino acid sequence SEQ ID NO.4, nucleotide sequence SEQ ID NO.230) were inserted at the dpp1 site, using the gRNA-N20 sequence CCAGGGATATCTCCGAATAG (SEQ ID NO.455) to construct the corresponding gRNA-2 plasmid. One copy of mevalonate kinase ERG12 (amino acid sequence SEQ ID NO.5, nucleotide sequence SEQ ID NO.231) and one copy of mevalonate-5-phosphate kinase ERG8 (amino acid sequence SEQ ID NO.6, nucleotide sequence SEQ ID NO.232) were inserted at the ho site on the genome, using the gRNA-N20 sequence GCCGGCTTGATCGACTCAGA (SEQ ID NO.456) to construct the corresponding gRNA-3 plasmid. One copy of mevalonate-5-pyrophosphate decarboxylase ERG19 (amino acid sequence SEQ ID NO.7, nucleotide sequence SEQ ID NO.233) and one copy of isopentenyl pyrophosphate isomerase IDI1 (amino acid sequence SEQ ID NO.8, nucleotide sequence SEQ ID NO.234) were inserted at the gal80 site on the genome. The gRNA-N20 sequence used was ATAAGGCTGCTGCTGAACGT (SEQ ID NO.457), and the corresponding gRNA-4 plasmid was constructed.

[0231] Donor plasmids were constructed as follows: pYC1 containing the complete expression cassettes of ERG20 and tHMG1 and homologous arms upstream and downstream of the integration site lpp1; pYC2 containing the complete expression cassettes of ERG10 and ERG13 and homologous arms upstream and downstream of the integration site dpp1; pYC3 containing the complete expression cassettes of ERG12 and ERG8 and homologous arms upstream and downstream of the integration site ho; and pYC4 containing the complete expression cassettes of ERG19 and IDI1 and homologous arms upstream and downstream of the integration site gal80. Using the CEN.PK2-1C genome as a template, the required fragments were amplified using primers (see Table 1). After gel recovery using the Tiangen Gel Recovery Kit, these fragments were ligated into the linearized backbone fragment of the expression vector pUC19 digested with HindIII using the Novizan ClonExpress Multis One Step Cloning Kit (Vazyme, catalog number: C113-02) via homologous recombination. The reaction system and conditions were performed according to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed. After sequencing confirmation, donor plasmids pYC1, pYC2, pYC3, and pYC4 were obtained.

[0232] Table 1. Fragments and primers required for vector construction, and primer sequences.

[0233] Using primers PF1 / PR7 and plasmid pYC1 as a template, the Donor 1 fragment was amplified. This fragment, along with plasmids containing Cas9 and gRNA-1, was then transformed into competent cells of yeast strain CEN.PK2-1C using the PEG / LiAC method, yielding strain YC1. Next, using primers PF8 / PR14 and plasmid pYC2 as a template, the Donor 2 fragment was amplified. This fragment, along with plasmids containing Cas9 and gRNA-2, was then transformed into competent cells of yeast strain YC1 using the PEG / LiAC method, yielding strain YC2. Finally, using primers PF15 / PR21 and plasmid pYC3 as a template, the Donor 3 fragment was amplified. This fragment, along with plasmids containing Cas9 and gRNA-3, was then transformed into competent cells of yeast strain YC2 using the PEG / LiAC method, yielding strain YC3. Using primers PF22 / PR28 and plasmid pYC4 as a template, the Donor4 fragment was amplified and transformed into competent cells of yeast strain YC3 via PEG / LiAC method, along with plasmids containing Cas9 and gRNA-4, to obtain strain YC4.

[0234] Construction of a lysine-based glycoside synthesis support (1) Preparation of target gene The geraniol geraniol pyrophosphate synthase gene BTS1, derived from Saccharomyces cerevisiae and provided by NCBI, was selected. Its amino acid sequence is shown in SEQ ID NO.9 and its nucleotide sequence is SEQ ID NO.235. It can be directly amplified from the Saccharomyces cerevisiae genome.

[0235] Selected from sage (sweet citrus) provided by NCBI Salvia sclarea The amino acid sequence of the SsLPPS gene (GenBank accession number: JQ478434.1), encoding lysine pyrophosphate diol ester synthase, was obtained by removing the signal peptide sequence of the first 63 amino acids, as shown in SEQ ID NO. 10. After codon optimization, the gene was synthesized using Genewiz, and its nucleotide sequence is shown in SEQ ID NO. 236. This nucleotide sequence was then inserted into the plasmid pUC-GW-kan (purchased from Genewiz) to obtain the plasmid pUC-GW-kan-tSsLPPS. As provided in the exemplary lysine-2-diol biosynthesis pathway ( Figure 1 Labdenediol pyrophosphate (LPP) is a direct precursor for the production of the target product, Labdenediol. Therefore, this invention aims to discover effective phosphorylases to dephosphate LPP and generate Labdenediol. Twenty-six phosphorylases derived from *Saccharomyces cerevisiae* (Saccharomyces cerevisiae) provided by NCBI were selected, with amino acid sequences as shown in SEQ ID NO. 11-216 and nucleotide sequences as shown in SEQ ID NO. 237-442. In addition to endogenous phosphorylases from *Saccharomyces cerevisiae*, enzyme genes from other species were also selected, including those derived from... Escherichia coli Phosphatidylglycerol phosphatase PgpA (amino acid sequence SEQ ID NO. 217), phosphatidylglycerol phosphatase PgpB (amino acid sequence SEQ ID NO. 218), phosphatidylglycerol phosphatase PgpC (amino acid sequence SEQ ID NO. 219), carbapenyl bisphosphatase YbjG (amino acid sequence SEQ ID NO. 220), and phosphatidylglycerol phosphatase derived from K12 Homo sapiens The dihydroxy diphosphatase DOLPP1 (amino acid sequence SEQ ID NO. 221) and phosphatase PLPP6 (amino acid sequence SEQ ID NO. 222) are derived from... Bacillus subtilis The farnesyl diphosphatase YisP (amino acid sequence SEQ ID NO. 223), or derived from... Phyllostachys edulis Farnesol synthase TPS2 (amino acid sequence SEQ ID NO. 224), or derived from... Zea maysThe noncyclic sesquiterpene synthase TPS1 (amino acid sequence SEQ ID NO. 225), or derived from... Oryza sativa subsp. japonica The farnesol synthase TPS13 (amino acid sequence SEQ ID NO. 226) was synthesized using Genewiz after codon optimization. Its nucleotide sequence is shown in SEQ ID NO. 443~SEQ ID NO. 452. This synthesized enzyme was then inserted into plasmid pUC-GW-kan (purchased from Genewiz) to obtain plasmids pUC-GW-kan-PgpA, pUC-GW-kan-PgpB, pUC-GW-kan-PgpC, pUC-GW-kan-YbjG, pUC-GW-kan-DOLPP1, pUC-GW-kan-PLPP6, pUC-GW-kan-YisP, pUC-GW-kan-TPS2, pUC-GW-kan-TPS1, and pUC-GW-kan-TPS13.

[0236] (2) Construction of recombinant plasmid for the synthesis of lysine-3-enediol This invention integrates the geraniol pyrophosphate synthase gene BTS1 into the dit1 site of the genome, using the gRNA-N20 sequence AAAGATACTCGTTTCGATAT (SEQ ID NO. 514). The truncated lysine pyrophosphate diol ester synthase encoding gene tSsLPPS is integrated into the gal2 site of the genome, using the gRNA-N20 sequence ATCCCCACGTTATTTATGTG (SEQ ID NO. 515). Candidate phosphatases are integrated into the adh5 site of the genome, using the gRNA-N20 sequence TCCAACACAAGATCCAACGA (SEQ ID NO. 516). The gRNA expression cassettes required for these three integration sites are sequentially tandemly constructed into the same vector, ultimately obtaining a tandem gRNA plasmid capable of editing three sites (dit1, gal2, and adh5) at once, named pYC-3gRNA. The nucleotide sequence of this recombinant plasmid is shown in SEQ ID NO. 453.

[0237] First, using the genome of Saccharomyces cerevisiae CEN.PK2-1C as a template, PCR amplification was performed using specific gene primers for PF29 / PR29, PF30 / PR30, PF31 / PR31, PF32 / PR32, and PF33 / PR33 (PCR system using 2×Phanta MaxMaster Mix, (Dye Plus), Vazyme). The upstream homologous arm dit1-US, promoter pGAL1 fragment, geranylgeranyl pyrophosphate synthase gene BTS1 fragment, terminator T-BTS1 fragment, and downstream homologous arm dit1-DS of the dit1 integration site were obtained. After gel recovery using the Tiangen Gel Recovery Kit, these fragments were ligated into the linearized backbone fragment of the expression vector pUC19 digested with HindIII. The ligation was performed using the Novizan ClonExpressMultis One Step Cloning Kit (Vazyme, catalog number: C113-02), following the kit's instructions. After seamless assembly, the cells were transformed into Trans1 T1 competent cells. Following sequencing confirmation, a donor plasmid containing the complete BTS1 expression cassette and the upstream and downstream homologous arms of the integration site dit1 was obtained and named pYC5.

[0238] Using plasmid pUC-GW-kan-tSsLPPS as a template, PCR amplification was performed using primer pair PF36 / PR36 (PCR system: 2×Phanta Max Master Mix, (Dye Plus), Vazyme) to obtain a truncated SsLPPS fragment encoding the lysine pyrophosphate synthase gene. Using *Saccharomyces cerevisiae* CEN.PK2-1C as a template, PCR amplification was performed using specific gene primer pairs PF34 / PR34, PF35 / PR35, PF37 / PR37, and PF38 / PR38, respectively, to obtain the upstream homologous arm fragment gal2-US, the promoter pGAL1 fragment, the terminator tADH1 fragment, and the downstream homologous arm fragment gal2-DS of the gal2 integration site. These fragments were recovered using a Tiangen gel extraction kit and ligated into the HindIII-digested linearized vector pUC19, followed by seamless assembly and transformation into Trans1 T1 competent cells. After sequencing confirmed that there were no errors, a donor plasmid containing the complete tSsLPPS expression cassette and the upstream and downstream homologous arms of the integration site gal2 was obtained and named pYC6.

[0239] For the construction of candidate phosphatase gene donor plasmids (e.g.) Figure 3As shown, candidate phosphatolases were integrated into the adh5 site on the genome. Using the *Saccharomyces cerevisiae* CEN.PK2-1C genome as a template, PCR amplification was performed using specific gene primers PF39 / PR39, PF40 / PR40, PF41 / PR41, and PF42 / PR42, respectively, to obtain the upstream homologous arm fragment adh5-US, the promoter pGAL1 fragment, the terminator tPGK1 fragment, and the downstream homologous arm fragment adh5-DS of the integration site adh5. For the construction of recombinant plasmids of endogenous candidate phosphatolases (i.e., SEQ ID NO. 237~442) in *Saccharomyces cerevisiae*, gene fragments were obtained by amplification using the *Saccharomyces cerevisiae* CEN.PK2-1C genome as a template. For the construction of recombinant plasmids for exogenous phosphorylase candidate enzymes, plasmids pUC-GW-kan-PgpA, pUC-GW-kan-PgpB, pUC-GW-kan-PgpC, pUC-GW-kan-YbjG, pUC-GW-kan-DOLPP1, pUC-GW-kan-PLPP6, pUC-GW-kan-YisP, pUC-GW-kan-TPS2, pUC-GW-kan-TPS1, and pUC-GW-kan-TPS13 were used as templates for amplification to obtain exogenous candidate phosphorylase gene fragments (i.e., SEQ ID NO.443~452). After gel recovery of the above fragments using the Tiangen Gel Recovery Kit, the upstream and downstream homologous arm fragments adh5-US and adh5-DS, the promoter fragment pGAL1, and the terminator fragment tPGK1 were ligated, respectively, with 216 different phosphorylase gene fragments into the HindIII-digested linearized vector pUC19. Homologous recombination was then performed using the Novizan Seamless Assembly Kit (ClonExpress Multis One Step Cloning Kit, Vazyme, catalog number: C113-02). The reaction system and conditions were performed according to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed. After sequencing confirmation, donor plasmids containing complete expression cassettes of different phosphorylases and the integration site adh5 upstream and downstream homologous arms were obtained and named pYC7~pYC222, respectively.

[0240] Table 2 shows the required primers and their sequences.

[0241] Construction of Lysine-containing diol synthesizing strain and well plate fermentation Using primers PF29 / PR33 and plasmid pYC5 as a template, the Donor5 fragment, namely dit1-US-pGAL1-BTS1-T-BTS1-dit1-DS, was amplified; using plasmid pYC6 as a template, the Donor6 fragment, namely gal2-US-pGAL1-tSsLPP1-tADH1-gal2-DS, was amplified; and using plasmids pYC7 to pYC222 as templates, 216 Donor DNA fragments containing different candidate phosphorylase expression cassettes, namely adh5-US-pGAL1-Phosphohydrolase-tPGK1-adh5-DS, were amplified and named Donor7 to Donor222, respectively. Donor7~Donor222, along with Donor5 fragment, Donor6 fragment, plasmid containing Cas9, and pYC-3gRNA plasmid, were transformed into competent cells of yeast strain YC4 using the PEG / LiAC method to obtain strains YC5~YC220.

[0242] After the above-mentioned strains YC5~YC220 were cultured in well plates and fermented according to Example 1, the samples were analyzed by gas chromatography. The gas chromatogram of the lysine diol standard is shown below. Figure 4 As shown, the peak time of lysine diol was 6.080 min. After analysis of the fermentation samples, 36 strains were found to have a peak at 6.080 min. Taking the gas chromatogram of sample strain YC6 as an example (…), Figure 5 It was inferred that the product lysantandrindiol had been formed, with a maximum concentration of 120 mg / L. Further mass spectrometry identification confirmed it as the target product lysantandrindiol. Figure 6 This indicates the successful discovery of an effective phosphorylase capable of converting the precursor lysandrindiol pyrophosphate (LPP) into the target product, lysandrindiol. Information on phosphorylases capable of successfully converting LPP to lysandrindiol is listed in Table 3. The specific family classifications of these phosphorylases are listed in Table 4.

[0243] Table 3. Fermentation levels of lysine-containing plate-grown phosphorylase strains

[0244] Table 4. Classification of Phosphohydrolases

[0245] The production of lysine diol was carried out using strain YC6. Specifically, a single colony of engineered strain YC6 was picked from a YPD plate and inoculated into a shake flask containing 20 ml of YPD. The culture was incubated at 30°C and 180 rpm for 48 hours to obtain a yeast seed culture. The engineered yeast seed culture was then inoculated at a 5% (v / v) inoculation rate into a 96-well plate containing fermentation medium LGM and incubated at 30°C in a deep-well plate shaker for 48 hours. After incubation, 4 times the fermentation volume of ethyl acetate was added for extraction. The upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. Approximately 120 mg / L of lysine diol was detected.

[0246] Example 3: Conversion of lysine-2-benzenediol or styracil-2-ol using fungus ATCC 20624. A single colony of fungus ATCC 20624 was picked from a YM plate and transferred to a shake flask containing 20 ml of seed culture medium YM. The culture was then incubated at 25°C and 180 rpm for 48 hours to obtain the seed culture of fungus ATCC 20624.

[0247] Lysine-containing diol at a final concentration of 8.9 g / L was added to the fermentation medium LGM, and then dispensed into 96-well plates. ATCC 20624 seed culture was inoculated at a rate of 5% (v / v) into 96-well plates containing the fermentation medium and lysine-containing diol, and cultured at 1000 rpm and 25°C for 72 hours. After culture, ethyl acetate (4 times the fermentation volume) was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the method described in Example 1. The gas chromatography results of the fermentation broth extract were compared with those of the lysine lactone and lysine diol standards, confirming that the main products were lysine diol and lysine lactone. Figure 7 As shown, the yield of perillaldehyde was 3.2 g / L, and the yield of perillaldehyde lactone was 0.08 g / L.

[0248] 5.4 g / L of perillyl alcohol was added to the fermentation medium LGM, and then dispensed into 96-well plates. ATCC20624 seed culture was inoculated at a 5% (v / v) inoculation rate into 96-well plates containing both fermentation medium and perillyl alcohol, and cultured at 1000 rpm and 25°C for 72 hours. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. The gas chromatography results of the fermentation broth extract were compared with those of perillyl lactone and perillyl diol standards to confirm that the products included perillyl diol and perillyl lactone. Figure 7As shown, the yield of perillaldehyde was 3.4 g / L, and the yield of perillaldehyde lactone was 0.68 g / L.

[0249] Example 4: Using fungus ATCC 20918 to convert lysine-1-diol or perillyl alcohol to perillyl lactone A single colony of fungus ATCC 20918 was picked from a YM plate and transferred to a shake flask containing 20 ml of seed culture medium YM. The culture was then incubated at 25°C and 180 rpm for 48 hours to obtain the seed culture of fungus ATCC 20918.

[0250] Lysine diol to a final concentration of 8.9 g / L was added to the fermentation medium LGM, and then dispensed into 96-well plates. ATCC 20918 seed culture was inoculated into the above 96-well plates at a 5% (v / v) inoculum and cultured at 1000 rpm and 25°C for 72 hours. After culture, ethyl acetate (4 times the fermentation volume) was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method shown in Example 1. The gas chromatography results of the fermentation broth extract were compared with those of the lysine lactone and lysine diol standards to confirm that the product was lysine lactone. Figure 8 As shown, the yield of perillaldehyde was 5.2 g / L.

[0251] Perillyl alcohol was added to the fermentation medium LGM to a final concentration of 5.4 g / L, and then dispensed into 96-well plates. ATCC 20918 seed culture was inoculated into the 96-well plates at a 5% (v / v) inoculum and cultured at 1000 rpm and 25°C for 72 hours. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction. The upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. The gas chromatography results of the fermentation broth extract were compared with those of perillyl lactone and perillyl diol standards to confirm that the product was perillyl lactone. Figure 8 As shown, the yield of perillaldehyde was 3.5 g / L.

[0252] Example 5: Production of perillaldehyde and perillyl lactone by co-culturing genetically engineered Saccharomyces cerevisiae and fungus ATCC 20624. Amplification culture of fungal ATCC 20624 seed culture. Single ATCC 20624 clones were picked from YM plates and inoculated into shake flasks containing 20 ml of seed culture medium YM. The flasks were incubated at 25°C and 180 rpm for 48 hours to obtain ATCC 20624 seed culture.

[0253] Amplification and culture of the engineered Saccharomyces cerevisiae YC6 seed culture. Single clones of engineered YC6 were picked from YPD plates and inoculated into shake flasks containing 20 ml of YPD. The cultures were then incubated at 30°C and 180 rpm for 48 hours to obtain the yeast seed culture.

[0254] The ATCC 20624 seed culture and the engineered strain YC6 seed culture were mixed at ratios of 1:8.5, 1:3.9, 1:1.55, 1:0.78, 2.58:1, 6.45:1, and 14:1, respectively, to obtain OD values. 600 The volumes (corresponding to the volumes shown in Table 5, with a total volume of 24 μL for both strains) were added to 96-well plates containing LGM fermentation medium and cultured at 25°C and 1000 rpm for 72 h in a deep-well plate shaker. Additionally, only the seed culture of engineered strain YC6 was added to the same 96-well plate for simultaneous culture as a negative control. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. As shown in Table 5, engineered strain YC6 alone produced 118 mg / L of lysine-enriched diol. When ATCC20624 was co-cultured with engineered strain YC6, the products perillaldehyde and perillyl lactone were detected. This phenomenon is consistent with the results when fungus ATCC 20624 alone transformed lysine-enriched diol, both producing alcohol and lactone products. The ratio of ATCC 20624 to engineered bacteria significantly affected the yield of perillaldehyde or perillyl lactone. At 25℃, the OD values ​​of the initial addition amounts of ATCC 20624 and engineered bacteria YC6 were [data missing]. 600 After the ratio was set to 6.45:1, especially the initial OD of ATCC20624 and engineered bacteria YC6, 600 At a ratio of 2.58:1, perillaldehyde was produced, with a yield of 6.29 mg / L. As the amount of ATCC20624 added decreased, the yields of perillaldehyde and perillaldehyde showed a trend of first increasing and then decreasing. Specifically, the OD values ​​of ATCC 20624 and the engineered bacteria YC6 at the initial addition amount... 600 At a ratio of 1:3.9, the yields of perillaldehyde and perillyl lactone were the highest, at 45.9 mg / L and 32.1 mg / L, respectively. Furthermore, the initial addition amounts of ATCC 20624 and engineered bacteria YC6 showed different OD values. 600 Only styraciol was detected at a ratio of 2.58:1 or 1:8.5; the initial addition amount of ATCC 20624 and engineered bacteria YC6 was [OD value missing]. 600 At a ratio of 1:0.78, only perillaldehyde was detectable. The initial OD values ​​of ATCC 20624 and engineered bacteria YC6 were [not specified]. 600When the ratio is 1:3.9 or 1:1.55, both perillaldehyde and perillyl glycol can be detected simultaneously. By adjusting the initial addition amount of ATCC 20624 and engineered bacteria YC6, mixtures of perillaldehyde and perillyl glycol in different ratios can be obtained, as well as single perillaldehyde or perillyl glycol.

[0255] Table 5. Effects of different addition ratios at 25℃ on the co-culture of engineered bacteria YC6 and fungus ATCC 20624.

[0256] Example 6: Production of perillaldehyde by co-culturing genetically engineered Saccharomyces cerevisiae and fungus ATCC 20918 Amplification culture of fungal ATCC 20918 seed culture. Single ATCC 20918 clones were picked from YM plates and inoculated into shake flasks containing 20 ml of seed culture medium YM. The flasks were then cultured at 25°C and 180 rpm for 48 hours to obtain ATCC 20918 seed culture.

[0257] Amplification and culture of the engineered Saccharomyces cerevisiae YC6 seed culture. Single clones of engineered YC6 were picked from YPD plates and inoculated into shake flasks containing 20 ml of YPD. The cultures were then incubated at 30°C and 180 rpm for 48 hours to obtain the yeast seed culture.

[0258] The ATCC 20918 seed culture and the engineered strain YC6 seed culture were mixed at ratios of 1:10.5, 1:4.8, 1:1.9, 1:0.96, 2.1:1, 5.2:1, and 11.5:1. 600 The corresponding volumes (as shown in Table 6, with a total added volume of 24 μL for both strains) were added to 96-well plates containing fermentation medium and cultured at 25°C and 1000 rpm for 72 h in a deep-well plate shaker. Additionally, only the seed culture of engineered strain YC6 was added to the same 96-well plate for simultaneous culture as a negative control. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. As shown in Table 6, engineered strain YC6 alone produced 118 mg / L of lysine-enriched diol. When ATCC 20918 was co-cultured with engineered strain YC6, only perillaldehyde was detected, consistent with the results when ATCC 20918 alone converted lysine-enriched diol, where only perillaldehyde was detected. The different ratios of ATCC 20918 and engineered strain YC6 significantly affected the yield of perillaldehyde. At 25℃, the OD of ATCC 20918 and engineered bacteria YC6 at initial addition levels 600The highest yield of perillaldehyde (77.4 mg / L) was observed at a ratio of 1:10.5. Increasing the amount of ATCC20918 added further reduced the yield of perillaldehyde.

[0259] Table 6. Effects of different addition ratios at 25℃ on the co-culture of engineered bacteria YC6 and fungus ATCC 20918.

[0260] Example 7: The effect of temperature on the products of mixed bacterial co-culture Following the methods described in Examples 5 and 6, fungi ATCC 20624, ATCC 20918, and the genetically engineered yeast YC6 were amplified and cultured to obtain seed culture.

[0261] Following the method described in Example 5, fungus ATCC 20624 and engineered yeast YC6 were co-cultured. Only the seed culture of engineered yeast YC6 was added to the same 96-well plate as a negative control. Keeping all other conditions unchanged, the culture temperature was adjusted from 25°C to 30°C. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. Figure 9 As shown, when the engineered strain YC6 was cultured alone at 30℃, it produced 217 mg / L of lysine-enriched diol, which was significantly higher than the yield at 25℃. With increasing ATCC 20624 addition, the yield of perillaldehyde showed a trend of first increasing and then decreasing. Increasing the temperature was also beneficial for the co-culture of ATCC 20624 and engineered strain YC6 to produce perillaldehyde. At 30℃, the highest perillaldehyde yield (159.4 mg / L) was achieved when the initial OD ratio of ATCC 20624 to engineered strain YC6 was 1:1.55, which was 3.47 times the yield under the optimal conditions at 25℃. Furthermore, trace amounts of perillyl lactone (approximately 5.3 mg / L) were detected. At initial OD ratios of ATCC 20624 to engineered strain YC6 of 2.58:1 or 6.45:1, only perillaldehyde was detected. By controlling the initial addition ratio of ATCC 20624 to engineered bacteria YC6 at different temperatures, mixtures of perillaldehyde and perillyl glycol in different proportions can be obtained, as well as single perillyl glycol.

[0262] Following the method described in Example 6, the fungus ATCC 20918 and the genetically engineered yeast YC6 were co-cultured. Only the seed culture of the engineered yeast YC6 was added to the same 96-well plate as a negative control. Keeping all other conditions unchanged, the culture temperature was adjusted from 25°C to 30°C. After culture, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method described in Example 1. Figure 9 As shown, at 30℃, the OD values ​​of ATCC 20918 and the initial amount of engineered bacteria added... 600 At a ratio of 1:1.9, the yield of perillaldehyde was the highest, reaching 57.8 mg / L, with an additional 27.6 mg / L of perillyl glycol. This result is lower than the yield of perillaldehyde under the optimal condition of 25℃, indicating that higher temperatures are unfavorable for the co-cultivation of ATCC 20918 with engineered bacteria to produce perillaldehyde. Furthermore, the increased temperature altered the product distribution, changing the production from solely perillaldehyde at various addition ratios under 25℃ to the production of both perillyl glycol and perillaldehyde at certain ratios. For example, the initial addition ratio of fungus ATCC 20918 to the engineered yeast YC6 was OD... 600 A certain amount of perillaldehyde and perillyl lactone can be produced at ratios of 1:1.9 or 1:0.96; at OD... 600 Only perillaldehyde can be detected at ratios of 2.1:1, 5.2:1, or 11.5:1. The specific growth rates of ATCC 20624, ATCC 20918, and genetically engineered bacteria differ at different temperatures, potentially leading to variations in the optimal mixed bacterial ratio and drastic changes in yield.

[0263] Example 8: Effects of different mixed culture methods of fungus ATCC 20624 and engineered yeast on the yield of perillaldehyde and perillyl lactone Following the methods described in Examples 5 and 6, the fungus ATCC 20624 and the genetically engineered yeast YC6 were amplified and cultured to obtain seed culture.

[0264] Yeast engineered strain seed culture was inoculated at a 5% (v / v) inoculation rate into 96-well plates containing fermentation medium LGM and cultured at 25°C in a deep-well plate shaker for 48 h. Fungal ATCC 20624 was inoculated into shake flasks containing 20 mL LGM and 0.1 g / L lysine-containing glycol and cultured at 25°C and 180 rpm for 48 h. ATCC 20624 induced with lysine-containing glycol was then added to wells of engineered strain YC6, such that the OD values ​​of ATCC 20624 and engineered strain YC6 were equal. 600The ratios were 2:1, 1:1, and 1:2. An additional 20 g / L glucose solution was added to the mixed-culture plates to provide the carbon source needed for further growth of the strain. The mixed-culture plates containing ATCC 20624 and engineered yeast YC6 were incubated at 25°C and 1000 rpm in a deep-well plate shaker for 72 h. After incubation, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method shown in Example 1. The results showed that co-culturing ATCC 20624 fungus with engineered yeast in a fermentation plate resulted in a higher OD value for both. 600 At a ratio of 1:2, the yields of perillaldehyde and perillyl lactone were the highest, at 50 mg / L and 21 mg / L, respectively. Under the above culture conditions, changing the culture temperature to 30℃, or omitting the addition of glucose solution, or omitting the inducer lysine-2-diol, did not yield higher yields of perillaldehyde and perillyl lactone.

[0265] Example 9: Effects of different mixed culture methods of fungus ATCC 20918 and engineered yeast on the yield of perilla lactone Following the methods described in Examples 5 and 6, the fungus ATCC 20918 and the genetically engineered yeast YC6 were amplified and cultured to obtain seed culture.

[0266] Yeast engineered strain seed culture was inoculated at a 5% (v / v) inoculation rate into 96-well plates containing fermentation medium LGM and cultured at 30°C in a deep-well plate shaker for 48 h. Fungus ATCC 20918 was inoculated into shake flasks containing 20 mL LGM and 0.1 g / L lysine-containing glycol and cultured at 25°C and 180 rpm for 48 h. ATCC 20918 induced with lysine-containing glycol was then added to wells of engineered strain YC6, allowing the OD values ​​of ATCC 20918 and engineered strain YC6 to be equalized. 600 The ratios were 2:1, 1:1, and 1:2. An additional 20 g / L glucose solution was added to the mixed-culture plates to provide the carbon source needed for further growth of the strain. The mixed-culture plates containing ATCC 20918 and engineered yeast YC6 were incubated at 30°C and 1000 rpm in a deep-well plate shaker for 72 h. After incubation, 4 times the fermentation volume of ethyl acetate was added for extraction, and the upper organic phase was collected by centrifugation and analyzed according to the analytical method shown in Example 1. The results showed that co-culturing ATCC 20918 fungus with engineered yeast in a fermentation plate resulted in a higher OD value for both. 600The highest yield of perillaldehyde (172 mg / L) was obtained at a ratio of 2:1. Under the above culture conditions, changing the culture temperature to 25℃, omitting the addition of glucose solution, or omitting the inducer lysine-2-diol did not yield higher yields of perillaldehyde.

[0267] The sequences used in the examples: SEQ ID NO. 1, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomycescerevisiae ERG20, amino acid sequence List MASEKEIRRERFLNVFPKLVEELNASLLAYGMPKEACDWYAHSLNYNTPGGKLNRGLSVVDTYAILSNKTVEQLGQEEYEKVAILGWCIELLQAYFLVADDMMDKSITRRGQPCWYKVPEVGEIAINDAFMLEAAIYKLLKSHFRNEKYYIDITELFHEVTFQTELGQLMDLITAP EDKVDLSKFSLKKHSFIVTFKTAYYSFYLPVALAMYVAGITDEKDLKQARDVLIPLGEYFQIQDDYLDCFGTPEQIGKIGTDIQDNKCSWVINKALELASAEQRKTLDENYGKKDSVAEAKCKKIFNDLKIDQLYHEYEESVAKDLKAKISQVDESRGFKADVLTAFLNKVYKRSK SEQ ID NO.2, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae) Saccharomyces cerevisiae tHMG1, amino acid sequence List MAADQLVKTEVTKKSFTAPVQKASTPVLTNKTVISGSKVKSLSSAQSSSSGPSSSSEEDDSRDIESLDKKIRPLEELEALLSSGNTKQLKNKEVAALVIHGKLPLYALEKKLGDTTRAVAVRRKALSILAEAPVLASDRLPYKNYDYDRVFGACCENVIGYMPLPVGVIGPLVIDGTSYHIPMATTEGCLVASAMRGCCKAINAGGGATTVLTKDGMTRGPVVRFPTLKRSGACKIWLDSEEGQNAIKKAFNSTSRFARLQHIQTCLAGDLLFMRFRTTTGDAMGMNMISKGVEYSLKQMVEEYGWEDMEVVSVSGNYCTDKKPAAINWIEGRGKSVVAETATIPGDVVRKVLKSDVSALVELNIAKNLVGSAMAGSVGGFNAHAANLVTAVFLALGQDPAQNVESSNCITLMKEVDGDLRISVSMPSIEVTGGGTVLEPQGAMLDLLGVRGPHATAPGTNARQLARIVACAVLAGELSLCAALAAGHLVQSHMTHNRKPAEPTKPNNLDATDINRLKDGSVTCIKS SEQ ID NO.3, derived from brewer's yeast ( Saccharomyces cerevisiae ERG10, amino acid sequence List MSQNVYIVSTARTPIGSFQGSLSSKTAVELGAVALKGALAKVPELDASKDFDEIIFGNVLSANLGQAPARQVALAAGLSNHIVASTVNKVCASAMKAIILGAQSIKCGNADVVVAGGCESMTNAPYYMPAARAGAKFGQTVLVDGVERDGLNDAYDGLAMGVHAEKCARDWDITREQQDNFAIESYQKSQKSQKEGKFDNEIVPVTIKGFRGKPDTQVTKDEEPARLHVEKLRSARTVFQKENGTVTAANASPINDGAAAVILVSEKVLKEKNLKPLAIKGWGEAAHQPADFTWAPSLAVPKALKHAGIEDINSVDYFEFNEAFSVVGLVNTKILKLDPSKVNVYGGAVALGHPLGCSGARVVVTLLSILQQEGGKIGVAAICNGGGGASSIVIEKI SEQ ID NO.4, derived from Saccharomyces cerevisiae (Saccharomyces cerevisia Saccharomyces cerevisiae ERG13, amino acid sequence List MKLSTKLCGIKGRLRPQKQQQLHNTNLQMTELKKQKTAEQKTRPQNVGIKGIQIYIPTQCVNQSELEKFDGVSQGKYTIGLGQTNMSFVNDREDIYSMSLTVLSKLIKSYNIDTNKIGRLEVGTETLIDKSKSVKSVLMQLFGENTDVEGIDTLNACYYGGTNALFNSLNWIESNAWDGRDAIVVCGDIAIYDKGAARPTGGAGTVAMWIGPDAPIVFDSVRASYMEHAYDFYKPDFTSEYPYVDGHFSLTCYVKALDQVYKSYSKKAISKLVSDPAGSDALNVLKYFDYNVFHVPTCKLVTKSYGRLLYNDFRANPQLFPEVDAELATRDYDESLTDKNIEKTFVNVAKPFHKERVAQSLIVPTNTGNMYTASVYAAFASLLNYVGSDDLQGKRVGLFSYGSGLAASLYSCKIVGDVQHIIKELDITNKLAKRITETPKDYEAAIELRENAHLKKNFKPQGSIEHLQSGVYYLTNIDDKFRRSYDVKK SEQ ID NO.5, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ERG12, amino acid sequence List MSLPFLTSAPGKVIIFGEHSAVYNKPAVAAASVSALRTYLLISESSAPDTIELDFPDISFNHKWSINDFNAITEDQVNSQKLAKAQQATDGLSQELVSLLDPLLAQLSESFHYHAAFCFLYMFVCLCPHAKNIKFSLKSTLPIGAGLGSSASISVSLALAMAYLGGLIGSNDLEKLSENDKHIVNQWAFIGEKCIHGTPSGIDNAVATYGNALLFEKDSHNGTINTNNFKFLDDFPAIPMILTYTRIPRSTKDLVARVRVLVTEKFPEVMKPILDAMGECALQGLEIMTKLSKCKGTDDEAVETNNELYEQLLELIRINHGLLVSIGVSHPGLELIKNLSDDLRIGSTKLTGAGGGGCSLTLLRRDITQEQIDSFKKKLQDDFSYETFETDLGGTGCCLLSAKNLNKDLKIKSLVFQLFENKTTTKQQIDDLLLPGNTNLPWTS SEQ ID NO.6, derived from brewer's yeast ( Saccharomyces cerevisiae ERG8, amino acid sequence MSELRAFSAPGKALLAGGYLVLDPKYEAFVVGLSARMHAVAHPYGSLQESDKFEVRVKSKQFKDGEWLYHISPKTGFIPVSIGGSKNPFIEKVIANVFSYFKPNMDDYCNRNLFVIDIFSDDAYHSQEDSVTEHRGNRRLSFHSHRIEEVPKTGLGSSAGLVTTALASFFVSDLENNVDKYREVIHNLSQVAHCQAQGKIGSGFDVAAAAYGSIRYRRFPPALISNLPDIGSATYGSKLAHLVNEEDWNITIKSNHLPSGLTLWMGDIKNGSETVKLVQKVKNWYDSHMPESLKIYTELDHANSRFMDGLSKLDRLHETHDDYSDQIFESLERNDCTCQKYPEITEVRDAVATIRRSFRKITKESGADIEPPVQTSLLDDCQTLKGVLTCLIPGAGGYDAIAVIAKQDVDLRAQTADDKRFSKVQWLDVTQADWGVRKEKDPETYLDK SEQ ID NO.7, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ERG19, amino acid sequence List MTVYTASVTAPVNIATLKYWGKRDTKLNLPTNSSISVTLSQDDLRTLTSAATAPEFERDTLWLNGEPHSIDNERTQNCLRDLRQLRKEMESKDASLPTLSQWKLHIVSENNFPTAAGLASSAAGFAALVSAIAKLYQLPQSTSEISRIARKGSGSACRSLFGGYVAWEMGKAEDGHDSMAVQIADSSDWPQMKACVLVVSDIKKDVSSTQGMQLTVATSELFKERIEHVVPKRFEVMRKAIVEKDFATFAKETMMDSNSFHATCLDSFPPIFYMNDTSKRIISWCHTINQFYGETIVAYTFDAGPNAVLYYLAENESKLFAFIYKLFGSVPGWDKKFTTEQLEAFNHQFESSNFTARELDLELQKDVARVILTQVGSGPQETNESLIDAKTGLPKE SEQ ID NO.8, derived from brewer's yeast ( Saccharomyces cerevisiae IDI1, amino acid sequence MTADNNSMPHGAVSSYAKLVQNQTPEDILEEFPEIIPLQQRPNTRSSETSNDESGETCFSGHDEEQIKLMNENCIVLDWDDNAIGAGTKKVCHLMENIEKGLLHRAFSVFIFNEQGELLLQQRATEKITFPDLWTNTCCSHPLCIDDELGLKGKLDDKIKGAITAAVRKLDHELGIPEDETKTRGKFHFLNRIHYMAPSNEPWGEHEIDYILFYKINAKENLTVNPNVNEVRDFKWVSPNDLKTMFADPSYKFTPWFKIICENYLFNWWEQLDDLSEVENDRQIHRML SEQ ID NO.9, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae BTS1, amino acid sequence MEAKIDELINNDPVWSSQNESLISKPYNHILLKPGKNFRLNLIVQINRVMNLPKDQLAIVSQIVELLHNSSLLIDDIEDNAPLRRGQTTSHLIFGVPSTINTANYMYFRAMQLVSQLTTKEPLYHNLITIFNEELINLHRGQGLDIYWRDFLPEIIPTQEMYLNMVMNKTGGLFRLTLRLMEALSPSSHHGHSLVPFINLLGIIYQIRDDYLNLKDFQMSSEKGFAEDITEGKLSFPIVHALNFTKTKGQTEQHNEILRILLLRTSDKDIKLKLIQILEFDTNSLAYTKNFINQLVNMIKNDNENKYLPDLASHSDTATNLHDELLYIIDHLSEL SEQ ID NO.10, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae tSsLPPS, amino acids sequence MASQASEKDISLVQTPHKVEVNEKIEESIEYVQNLLMTSGDGRISVSPYDTAVIALIKDLKGRDAPQFPSCLEWIAHHQLADGSWGDEFFCIYDRILNTLACVVALKSWNLHSDIIEKGVTYIKENVHKLKGANVEHRTAGFELVVPTFMQMATDLGIQDLPYDHPLIKEIADTKQQRLKEIPKDLVYQMPTNLLYSLEGLGDLEWERLLKLQSGNGSFLTSPSSTAAVLMHTKDEKCLKYIENALKNCDGGAPHTYPVDIFSRLWAIDRLQRLGISRFFQHEIKYFLDHIESVWEETGVFSGRYTKFSDIDDTSMGVRLLKMHGYDVDPNVLKHFKQQDGKFSCYIGQSVESASPMYNLYRAAQLRFPGEEVLEEATKFAFNFLQEMLVKDRLQERWVISDHLFDEIKLGLKMPWYATLPRVEAAYYLDHYAGSGDVWIGKSFYRMPEISNDTYKELAILDFNRCQTQHQLEWIHMQEWYDRCSLSEFGISKRELLRSYFLAAATIFEPERTQERLLWAKTRILSKMITSFVNISGTTLSLDYNFNGLDEIISSANEDQGLAGTLLATFHQLLDGFDIYTLHQLKHVWSQWFMKVQQGEGSGGEDAVLLANTLNICAGLNEDVLSNNEYTALSTLTNKICNRLAQIQDNKILQVVDGSIKDKELEQDMQALVKLVLQENGGAVDRNIRHTFLSVSKTFYYDAYHDDETTELHIFKVLFRPVV SEQ ID NO.11, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ADR1, amino acid sequence List SEQ ID NO.12, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae APP1, amino acid sequence List MNSQGYDESSSSTAATSGPTSGDPRMGKKQRFMNLIRTTKDVYIPNLTSSISQKTMDGIRSTTNSFEGYNDLPMELPHNTTITYFPTYTTTNLVDPDGLSAPRKDFETTVRCAVSYPGNPTSRRNRWLLSLCKQYLRTGTAEADVAPVVPPHLEEDSGDLNDSQSSIESSLSSKSENRYSHMGIQEEDVLNERIQGFLSKKVPNTPVVVDLLPKDKLRGDTASFFGTTDSYGNLLIKAETDFLPSKINITLDTPIEGHADPISETFPANYVSPYGIGLISDIDDTIKHTGVTGDRRSMFRNVFIHDVQSWVIDGVPLWYKTLHDVADVDFFYVSNSPIQTFTLLKQYICANFPPGPIFLKQYSGNFFSTIMTSSANRKIQPIANILKDFPKKKFILVGDSGEHDLEAYTTTALQFPNQILAIYIRCCSNSMSDVPSHDEEVMNEVNNIIELQQRPMQMTKSTVRTRRRPPPPPIPSTQKPSLTEEQTESIRMSRRNKDENNTKRVAPPPLPNRQLPNLDANTYYVPSSQNDYGMYGAFMDKKADEWKRRVMDSIQKLSNQDTTLMFFSDPALSLEDSIRRIREKYSN SEQ ID NO.13, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae ARR2, amino acid sequence List MVSFITSRQLKGLIENQRKDFQVVDLRREDFARDHITNAWHVPVTAQITEKQLNQLIKGLSDTFSSSQFVKVIFHCTGSKNRGPKVAAKFETYLQEEDITSKFESCILVGGFYAWETHCRESNLKLIVSG SEQ ID NO.14, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae BNI4, amino acid sequence ListMSDSVSDSKSSELLNSTFYSTSINTLDHARTFRNSLILKEISGQSLNSSIKPCESVLDRDVESSVLQRSFGDSNARDSEVQTVNMTTSPSLSALADILNERSKYADQKTRKAQNIESSIIEEEEEEAEEQNNSINYEDITGSRLSVREDANENLAMTSPNLIDIDGSNSIQVAPLSLPSFEEPDFLSTPRVKPDSQGPRSKVSTQRTILERDNNFPVKREENTIISSETESTTHSVPFLKEDPKPSPPSSKLYNPKVRLNKAEARKYTDSSAQRTSAGSVLEDTSMHKKKKSIFSFLKKKEPKPVIGNNSVTNEKNKMSSSSTFSMNIQTSLKTPEKLKKKSHSSSIFNSFLKGIETSDSPRKEPIRQKKRTPKSKDKKQDTEQIIDAASVLSTESPLLRKNHDDTPVKIDHVTRLIDQRKPTPLNMDLILGGDKQINTPLQ EHVKEDDDAKNDLQLPTKDNFLSLDYEAPSPAFSKHDTGEVLFPKFLDNHEVDSIVSLERTRSTKSNKRSSMNSQRRSLTDTLSIKAQSEGMFITEASSVVLSTPDLTKSPASSILKNGRFEYSDNFSREHSYEGTTNEDFLDIRDDSGPLKKDDIFLESIEQKFDQLVMASDEEKTEVERDVPKPREEPLKKDSERQSVYADDDNELISDIMEFASFINFGDDDLNLDLDGDTTASYATETPEVGNDEVNSDTFDARNNKEDSYKEKETQSYSAAGEATTYGDERQGQLHTFEQDGSEINDNEFENEDFNKHIEQPIEVTPRNNAYLPEFEPNRPVSMSFKGLKAPRMNTSFIDSMTPDSPVKSDLTSLGEVYVNSNNDQGVRFSSQIILYDTYGEFEYDRHPEISTCNQLTPQLAQMIKLELNELKSAMEVHDDSRCYTHFY SEQ ID NO.15, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae BSP1, amino acid sequence ListMTKYERDPELVNFLSKVEDLNSKRYSNIPSSKPAGEALSPVRSHNSGEYRRADMMTGKNVEGCDNLAYRSAYNYEMTFSPKKTHYSLSELNLERITPRPDLEGSASQKEKKFLISEEDYLLLQKLKASQTYNDSNADKNLPSFEKGPRMPSRGRPRPREKEIITIQYDFELPGRADIPSSSSSSSPPPLPTRRDHIKITDGNEEKPLLPTRPNKAEVTESPSSRSIKPDAVVPERVKPAPPVSRSTKPASFLSSLEDNKLTKAKSYNSEMETPKTTVKSSHIDYLDSI QLKPTTLSPTMKNKPKPTPPPSPPAKRIPRSESFIKSMLNSNLTTTSKPSLPEKPQKLRNANLAAHKTKPSIPPKKVELNIVLPELRPVETSPTKQNFENSIDLPKLRSSNRNIKKEEEDSIPEAIKGIQNLKKTKQQKPAIPQKKSFLTNNSKNTTLKNGDDINKLNDEIEALSLRNNLKRPPTAPQRKISLPEALRRKVELMKKSKTEPVLESSNELSINAKLDAIIASRNLRASNTLPELSGVNTNIATSDKYTTSRDETVKETKPLVHPNKNRTRGPRRKLPTRV SEQ ID NO.16, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae BUD14, amino acid sequence ListMSNKEEHVDETSASGVKEVSSIAARHDNGYAPSLITSTSGMDSFQSHALLNDPTLIEDYSDIINNRPTSGSKLTLGNEDSESMGGSVVVTPTSNKSSPFNSKLNILSNAAEKGHDVLRNRDDDKELEENVEKHMHSNSKRDQRHYKENSSELPDSYDYSDSEFEDNLERRLQEIET DSVDSADKDEVHFSVNNTMNPDVDDFSDGLKYAISEDEEENYSDDDDFDRKFQDSGFQGEKDDLEEENDDYQPLSPPRELDPDKLYALYAFNGHDSSHCQLGQDEPCILLNDQDAYWWLVKRITDGKIGFAPAEILETFPERLARLNCWKNENMSSQSVASSDSKDDSISSGNKN QSDAESIIPTPALNGYGGGNKSVSFNDVVGYADRFIDDAIEDTSLDSNDGGEGNGQSYDDDVDNDKETKVTHRDEYTEAKNLFGFQDDTSDVVSDVSFSTSLNTPLNVKKVRRQDNKNESEPKTSSSKDREDDYNANRYVGQEKSEPVDSDYDTDLKKVFEAPRMPFANGMAKS DSQNSLSTIGEFSPSSSEWTNESPSTPIVEESSSIPSSRAIKDISQYIHAKSKIEETTNVENTEGQIQASLGSSGGMANQTDAEQPKEELEKHHSTPEEEKQSTLSLHSSSEEDFYMDEQRAVSSASINSSLSGSRALSNTNMSDPASKPNSLVQHLYAPVFDRMDVLMKQLDEIRK SEQ ID NO.17, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CAX4, amino acid sequence List MNSTAAAINPNPNVIPFDDTYILYDSHDFLSFLSFLSAYFSLMPILVLAFYLSWFIITRELEACIVAFGQLMNEIFNNVIKNIIKQPRPVSFGASFQNDTIRSGYGMPSAHSQFMGFCFTYNSLKIYTSWKNLNFLEKCIFSGALALLSFCVCFSRVYLHYHNLDQVIVGFSVGALTGSLYFFIVGIIRELGLINWFLKLRIVRLFYMTDSYNLAPLTLKENYEAYWKRINQRSFNDKSKRD SEQ ID NO.18, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CDC14, amino acid sequence List MRRSVYLDNTIEFLRGRVYLGAYDYTPEDTDELVFFTVEDAIFYNSFHLDFGPMNIGHLYRFAVIFHEILNDPENANKAVVFYSSASTRQRANAACMLCCYMILVQAWTPHQVLQPLAQVDPPFMPFRDAGYSNADFEITIQDVVYGVWRAKEKGLIDLHSFNLESYEKYEHVEFGDFNVLTPDFIAFASPQEDHPKGYLATKSSHLNQPFKSVLNFFANNNVQLVVRLNSHLYNKKHFEDIGIQHLDLIFEDGTCPDLSIVKNFVGAAETIIKRGGKIAVHCKAGLGRTGCLIGAHLIYTYGFTANECIGFLRFIRPGMVVGPQQHWLYLHQNDFREWKYTTRISLKPSEAIGGLYPLISLEEYRLQKKKLKDDKRVAQNNIEGELRDLTMTPPSNGHGALSARNSSQPSTANNGSNSFKSSAVPQTSPGQPRKGQNGSNTIEDINNNRNPTSHANRKVVIESNNSDDESMQDTNGTSNHYPKVSRKKNDISSASSSRMEDNEPSATNINNAADDTILRQLLPKNRRVTSGRRTTSAAGGIRKISGSIKK SEQ ID NO.19, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CDC48, amino acid sequence ListMGEEHKPLLDASGVDPREEDKTATAILRRKKKDNMLLVDDAINDDNSVIAINSNTMDKLEFRGDTVLVKGKKRKDTVLIVLIDDELEDGACRINRVVRNNLRILRGDLVTIHPCPDIKYATRISVLPIADTIEGITGNLFDVFLKPYFVEAYRPVRKGDHFVVRGGMRQVEFKVVDVEPEEYAVVAQDTIIHWEGEPINREDEENNM NEVGYDDIGGCRKQMAQIREMVELPLRHPQLFKAIGIKPPRGVLMYGPPGTGKTLMARAVANETGAFFFLINGPEVMSKMAGESESNLRKAFEEAEKNAPAIIFIDEISIAPKRDKTNGEVERRVVSQLLTLMDGMKARSNVVVIAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEVLRIHTKNMKLADDVDLEALAAETHGYV GADIASLCEAAMQQIREKMDLIDLDEDEIDAEVLDSLGVTMDNFRFALGNSNPSALRETVVESVNVTWDDVGGLDEIKEELKETVEYPVLHPDQYTKFGLSPSKGVLFYGPPGTGKTLLAKAVATEVSANFISVKGPELLSMWYGESESNIRDIFDKARAAAPTVVFLDELDSIAKARGGSLGDAGGASDRVVNQLLTEMDGNMNAKKNVFVIGATNRPDQIDPAILRPGRLDQLIYVPLPDENARSILNAQLRKTPLEPGLELTAIAKATQGFSGADLLYIVQRAAKYAIKDSIEAHRQHEAEKEVKVEGEDVEMTDEGAKAEQEPEVDPVPYITKEHFAEAMKTAKRSVSDAELRRYEAYSQQMKASRGQFSNFNFNDAPLGTTATDNANSNNSAPSGAGAAFGSNAEEDDDLYS SEQ ID NO.20, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CDC55, amino acid sequence ListMAQNNFDFKFSQCFGDKADIVVTEADLITAVEFDYTGNYLATGDKGGRVVLFERSNSRHCEYKFLTEFQSHDAEFDYLKSLEIEEKINEIKWLRPTQRSHFLLSTNDKTIKLWKVYEKNIKLVSQNNLTEGVTFAKKGKPDNNHNSRGGSVRAVLSLQSLKLPQLSQHDKIIAATKPRIYSNAHTYHINSISLNSDQETFLSADDLRINLWNLDIPDQSFNIVDIKPTNMEELTEVITASAEFHPQECNLFMYSSSKGTIKLCDMRQNSLCDNKTFEEYLDPINHNFFETEITSSISDIKFSPNGRYIASRDYLTVKIWDVNMDNKPLKTINIHEQLKERLSDTYENDAIFDKFEVNFSGDSSSVMTGSYNNNFMIYPNVVTSGDNDNGIVKTFDEHNAPNSNSNKNIHNSIQNKDSSSNGHKRRSNGRNTGMVGSSNSSRSSIAGGEGANPEDSGTEMNEIVLQADKTAFRNKRYGSLAQRSARNKDWGDDIDFKKNILHFSWHPRENSIAVAATNNLFIFSAL SEQ ID NO.21, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CET1, amino acid sequence ListMSYTDNPPQTKRALSLDDLVNHDENEKVKLQKLSEAANGSRPFAENLESDINQTETGQAAPIDNYKESTGHGSHSQKPKSRKSSNDDEETDTDDEMGASGEINFDSEMDFDYDKQHRNLLSNGSPPMNDGSDANAKLEKPSDDSIHQNSKSDEEQRIPKQGNEGNIASNYITQVPLQKQKQKQTEKKIAGNAVGSVVKKEEANAAVDNIFEEKATLQSKKNNIKRDLEVLNEISASSKPSKYRNVPIWAQKWKPTIKALQSINVKDLKIDPSFLN IIPDDDLTKSVQDWVYATIYSIAPELRSFIELEMKFGVIIDAKGPDRVNPPVSSQCVFTELDAHLTPNIDASLFKELSKYIRGISEVTENTGKFSIIESQTRDSVYRVGLSTQRPRFLRMSTDIKTGRVGQFIEKRH VAQLLLYSPKDSYDVKISLNLELPVPDNDPPEKYKSQSPISERTKDRVSYIHNDSCTRIDITKVENHNQNSKSRQSETTHEVELEINTPALLNAFDNITNDSKEYASLIRTFLNNGTIIRRKLSSLSYEIFEGSKKVM SEQ ID NO.22, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CMD1, amino acid sequence List MSSNLTEEQIAEFKEAFALFDKDNNGSISSSELATVMRSLGLPSEAEVNDLMNEIDVDGNHQIEFSEFLALMSRQLKSNDSEQELLEAFKVFDKNGDGLISAAELKHVLTSIGEKLTDAEVDDMLREVSDGSGEINIQQFAALLSK SEQ ID NO.23, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CMP2, amino acid sequence ListMSSDAIRNTEQINAAIKIIENKTERPQSSTTPIDSKASTVAAANSTATETSRDLTQYTLDDGRVVSTNRRIMNKVPAITSHVPTDEELFQPNGIPRHEFLRDHFKREGKLSAAQAARIVTLATELFSKEPNLISVPAPITVCGDIHGQYFD LLKLFEVGGDPATTSYLFLGDYVDRGSFSFECLIYLYSLKLNFNDHFWLLRGNHECKHLTSYFTFKNEMLHKYNLDIYEKCCESFNNLPLAALMNGQYLCVHGGISPENLSQDINNLNRFREIPSHGLMCDLLWADPIEEYDEVLDKDLT EEDIVNSKTMVPHHGKMAPSRDMFVPNSVRGCSYAFTYRAACHFLQETGLLSIIRAHEAQDAGYRMYKNTKTLGFPSLLTLFSAPNYLDTYNNKAAILKYENNVMNIRQFNMTPHYWLPDFMDVFTWSLPFVGEKVTEMLVAILNICTEDELENDTPVIEELVGTDKKLPQAGKSEAPQPATSASPKHASILDDEHRRKALRNKILAVAKVSRMYSVLREETNKVQFLKDHNSGVLPRGALSNGVKGLDEALSTFERARKHDLINEKLPPSLDELKNENKKYYEKVWQKVHEHDAKNDSK SEQ ID NO.24, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CNA1, amino acid sequence ListMSKDLNSSRIKIIKPNDSYIKVDRKKDLTKYELENGKVISTKDRPIASVPAITGKIPSDEEVFDSKTGLPNHSFLREHFFHEGRLSKEQAIKILNMSTVALSKEPNLLKLKAPITICGDIHGQYYDLLKLFEVGGDPAEIDYLFLGDYVDRGAFSFECLIYLYSLKLNNLGRFWMLRGNHECKHLTSYFTFKNEMLHKYDMEVYDACCRSFNVLPLAALMNGQYFCVHGGISPELKSVEDVNKINRFREIPSRGLMCDLLWADPVENYDDARDGSE FDQSEDEFVPNSLRGCSFAFTFKASCKFLKANGLLSIIRAHEAQDAGYRMYKNNKVTGFPSLITMFSAPNYLDTYHNKAAVLKYEENVMNIRQFHMSPHPYWLPDFMDVFTWSLPFVGEKVTSMLVSILNICSEQELDPESEPKAAEETVKARANATKETGTPSDEKASSAILEDETRRKALRNKILAIAKVSRMFSVLREESEKVEYLKTMNAGVLPRGALARGTEGLNETLSTFEKARKEDLINEKLPPSLSEVEQEKIKYYEKILKGAEKKPQL SEQ ID NO.25, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CNB1, amino acid sequence List MKLDRDSSGSIDKNEFMSIPGVSSNPLAGRIMEVFDADNSGDVDFQEFITGLSIFSGRGSKDEKLRFAFKIYDIDKDGFISNGELFIVLKIMVGSNLDDEQLQQIVDRTIVENDSDDGGRLSFEEFKNAIETTEVAKSLTLQYDV SEQ ID NO.26, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CSS1, amino acid sequence ListMFNRLNKFQAALALALYSQSALGQYYSNSTSISSNSSSTSVVSSSSGSVSISSSIAETSSSATDILSSITQSASSTSGVSSSVGPSSSSVVSSSVSQSSSSVSDVSSSVSQSSSSASDVSSSVSQSASSTSDVSSSVSQSSSSASDVSSSVSQSSSSASDVSSSVSQSASSASDVSSSVSQSASSTSDVSSSVSQSSSSASDVSSSVSQSSSSASDVSSSVSQSASSTSDVSSSVSQSASSTSGVSSSGSQSVSSASGSSSSFPQSTSSASTASGSATSNSLSSITSSASSASATASNSLSSSDGTIYLPTTTISGDLTLTGKVIATEGVVVAAGAKLTLLDGDKYSFSADLKVYGDLLVKKSKETYPGTEFDISGENFDVTGNFNAEESAATSASIYSFTPSSFDNSGDISLSLSKSKKGEVTFSPYSNSGAFSFSNAILNGGSVSGLQRRDDTEGSVNNGEINLDNGSTYVIVEPVSGKGTVNIISGNLYLHYPDTFTGQTVVFKGEGVLAVDPTETNATPIPVVGYTGKNQIAITADITALSYDGTTGVLTATQGNRQFSFAIGTGFSSSDFSVSEGIFAGAYAYYLNYNGVVATSAASSSTASGASASVTGSTSFGASVTGSTASTSFGASVTGSTASTSFGASVTGSTSVYTTTLDYVNATSTVVVSCSETTDSNGNVYTITTTVPCSSTTATITSCDETGCHVSTSTGAVVTETVSSKSYTTATVTHCDDNGCNTKTVTSECSKETSATTASPKSYTTVTVTHCDDNGCNTKTVTSEAPEATTTTTVSSQSYTTATVTHCDDNGCKTKTVTSEAPEATTTTVSPKTYTTATVTQCDDNGCSTKTVTSECPEETSATTTSPKSYTTVTVTHCDDNGCNTKTVTSEAPEATTTTVSPKTYTTATVTQCDDNGCSTKTVTSEAPKETSETSETSAAPKDIHYCHWLLNGDDNGCNVKIITSKIPEATSTVTQLVLLQSHTLLSLLRVLKQPH SEQ ID NO.27, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CTK1, amino acid sequence List MSYNNGNTYSKSYSRNNKRPLFGKRSPNPQSLARPPPPKRIRTDSGYQSNMDNISSHRVNSNDQPGHTKSRGNNNLSRYNDTSFQTSSRYQGSRYNNNNTSYENRPKSIKRDETKAEFLSHLPKGPKSVEKSRYNNSSNTSNDIKNGYHASKYYNHKGQEGRSVIAKKVPVSVLTQQRSTSVYLRIMQVGEGTYGKVYKAKNTNTEKLVALKKLRLQGEREGFPITSIREIKLLQSFDHPNVSTIKEIMVESQKTVYMIFEYADNDLSGLLLNKEVQISHSQCKHLFKQLLLGMEYLHDNKILHRDVKGSNILIDNQGNLKITDFGLARKMNSRADYTNRVITLWYRPPELLLGTTNYGTEVDMWGCGCLLVELFNKTAIFQGSNELEQIESIFKIMGTPTINSWPTLYDMPWFFMIMPQQTTKYVNNFSEKFKSVLPSSKCLQLAINLLCYDQTKRFSATEALQSDYFKEEPKPEPLVLDGLVSCHEYEVKLARKQKRPNILSTNTNNKGNGNSNNNNNNNNDDDDK SEQ ID NO.28, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CTK2, amino acid sequence List MPSTFESQLFFSRPFLSKRQIQRAQKNTISDYRNYNQKKLAVFKFLSDLCVQLKFPRKTLETAVYFYQRYHLFNRFETEVCYTVATSCLTLGCKEVETIKKTNDICTLSLRLRNVVKINTDILENFKKRVFQIELRILESCSFDYRVNNYVHIDEYVIKIGRELSFDYKLCNLAWVIAYDALKLETILVIPQHSIALAILKIAYELLDNKNWSSKRYSLFETDEKSVNEAYFDIVNFYINSFDMCDLQRHLPADLLPIGVERFMELKKNAGPESGLPQIPDHLLNADPYITITRDNNVQERRYVLSLELINGESSINSSTRHA SEQ ID NO.29, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae CTK3, amino acid sequence ColumnMDSLEARLQFIQVLKNLQKTLHKTRDSITSSSTTTPPSSQQKLNNDPIQFYLRNYRHHYEDFHQCLFDTTMKMDPLDRLDVVIYYVRIIRNLYPHSHSNVTKVNLEVLLMDIDLVFELCLPCQDWKSLTNQATCKELFLDLSKLIH YDATSVTHTPSDTTLIDATTWYSVKTERTTKDYKESLQRTESLLKDRDLKKLAFFQQFNSDTTAINPDLQTQPTNANILLHRMEADRELHKRSKETSWYIERPSNDILDESEFKSLWTHFETTDSGFDKDDYKNIKALNDIAKASYMY SEQ ID NO.30, derived from brewer's yeast ( Saccharomyces cerevisiae CTL1, amino acid sequence Column MSDQPETPSNSRNSHENVGAKKADANVASKFRSLHISETTKPLTSTRALYKTTRNNSRGATEFHKHVCKLAWKYLACIDKSSISHIEIEMKFGVITDKRTHRRMTPHNKPFIVQNRNGRLVSNVPEQMFSSFQELLRSKSENPSKCAPRVVKQVQKYTKDSIYNCNNASKVGKLTSWRCEDLRNKELKLTYIKKVRVKDFLIRYPQSSLDAKISILEVPEYETSAAFRNGFILQRTKSRSTYTFNDKMPLHLDLTKVTTTTRNSHQYTSHEVEVEMDPIFKETISANDREKFNEYMCSFLNASDLIRKAAERDNMLTT SEQ ID NO.31, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae CTO1, amino acid sequence ColumnMKTIIISDFDETITRVDTICTIAKLPYLLNPRLKPEWGHFTKTYMDGYHKYKYNGTRSLPLLSSGVPTIISQSNFNKLFADELKYQNHNRVVELNSVNEITKQQIFKSISLDQMKTFARDQNHEDCLLRDGFKTFCSSVVKNFESDFYVLSINWSKEFIHEVIGDRRLKNSHIFCNDLKKVSDKCSQSYNGEFDCRLLTGSDKVKILGEILDKIDSGCNKEGNSCSYWYIGDSETDLLSILHPSTNGVLLINPQENPSKFIKITEKIIGIPKDKISSFEADNGPAWLQFCEKEGGKGAYLVKSWDSLKDLIMQVTKM SEQ ID NO.32, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae DCR2, amino acid sequence Column MIRLPRLYQRYLLYLLVVFVVIALFYFLQAPRVEEHIGFDLALPISHVDNLWFQNKGLEGFSNDDKLVVNIGYDECFHIGRFYEGCFNRHELKSTLTDGHQYLQRKRIHKDLRGSFGRRWFGKSEYLYYDVLYPALVDYFGSNLEKLNVEAVTGISKYPKDKSLPFMDVSITFEPISIELLQKRSYISDINILFGVDCIQPIANWTLQKEFPLVKYRYSEPAYLTYKFVGTRPVDTGAQRLQETDEGKFKIVQLADLHLGVGESECIDEYPKHEACKADPKTETFVQQVL DIEKPQLVVFTGDQIMGDRSIQDSETVLLKAVAPVIARKIPWAMVWGNHDDEGSLTRWQLSEIASVLPYSLFKFSPHDTHDNTFGVGNYIYQIFSNNDTEPVVGTLYFLDSHKYSTVGKIYPGYDWIKESQWKYIEDYHDVNLK FKTGLSMAFFHIPLPEYLNIESKTHPGEKNPLIGMYKEGVTAPKYNSEGITTLDRLSVDVVSCGHDHCNDYCLRDDSTPNKIWLCYGGGGGEGGYAGYGGTERRIRIYEINVNENNIHTWKRLNGSPKEIFDFQSMLDGNSPESV SEQ ID NO.33, derived from brewer's yeast ( Saccharomyces cerevisiae DDP1, amino acid sequence ColumnMGKTADNHGPVRSETAREGRENQVYSPVTGARLVAGCICLTPDKKQVLMITSSAHKKRWIVPKGGVEKDEPNYETTAQRETWEEAGCIGKIVANLGTVEDMRPPKDWNKDIKQFENSRKDSEVAKHPPRTEFHFYELEIENLLDKFPECHKRHRKLYSYTEAKQNLIDAKRPELLEALNRSAIIKDDK SEQ ID NO.34, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae DET1, amino acid sequence Column MCEENVHVSEDVAGSHGSFTNARPRLIVLIRHGESESNKNKEVNGYIPNHLISLTKTGQIQARQAGIDLRVLNVDDHNLVEDLAKKYIKDESSRRTLPLKDYTRLSREKDTNIVFYTSPYRRARETLKGILDVIDEYNELNSGVRICEDMRYDPHGKQKHAFWPRGLNNTGGVYENNEDNICEGKPGKCYLQYRVKDEPRIREQDFGNFQKINSMQDVMKKRSTYGHFFFRFPHGESAADVYDRVASFQETLFRHFHDRQERRPRDVVVLVTHGIYSRVFLMKWFRWTYEEFEFSFTNVPNGSVMVMELDESINRYVLRTVLPKWTDCEGDLTT SEQ ID NO.35, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae DIA3, amino acid sequence ColumnMVKPVIFAICLGVLLSKALSIPLRSFADIELIGSQKSLFPFLGGSAPYFSFPANYGIPTDIPEGCRLTQVQMIGRHGERYPTRSEAKDIFEVWYKISNYTGKYEGSLSFLNNGYEFFIPDESLLEMETTLQNSIDVLNPYTGEMNAKRHAREFLAKYGKLMENCTNFPIFTTNSKRIYDTAQYFAEALGDGFNISLQTLSENSSSGANTLAAKSSCPNWNSNANNDILMSYSRDYLENISDRLNDENKGLNLSRKDAAALFSWCAFELNAKGYSNICDIFSAAAELIHYSYETDLTSFYQNGPGYKLIKSIGANLFNATVKLIRQSAHLDQKVWLSFTHDTDILNYLTTAGLIDDTRNLTTNHVPFRDHSYHRSWYIPQGARVYTEKFQCSNDSYVRYVVNDAVVPIESCSSGPGFSCEEGTFFYEYAKDRLRGVSFYEDCDVSKVSKEKELTFYWDWNTTRYNASLVNQ SEQ ID NO.36, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae DOG1, amino acid sequence Column MAEFSDLCLFDLDGTIVSTTVAAEKAWTKLCYEYGVDPSELFKHSHGARTQEVLRRFFPKLDDTDNKGVLALEKDIAHSYLDTVSLIPGAENLLLSLDVDTETQKKLPERKWAIVTSGSPYLAFSWFETILKNVGKPKVFITGFDVKNGKPDPEGYSRARDLLRQDLQLTGKQDLKYVVFEDAPVGIKAGKAMGAITVGITSSYDKSVLFDAGADYVVCDLTQVSVVKNNENGIVIQVNNPLTRA SEQ ID NO.37, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae DOG2, amino acid sequence ColumnMPQFSVDLCLFDLDGTIVSTTTAAESAWKKLCRQHGVDPVELFKHSHGARSQEMMKKFFPKLDNTDNKGVLALEKDMADNYLDTVSLIPGAENLLLSLDVDTETQKKLPERKWAIVTSGSPYLAFSWFETILKNVGKPKVFITGFDVKNGKPPDPEGYSRARDLRQDLQLTGKQDLKYVVFEDAPVGIKAGKAMGAITVGITSSYDKSVLFDAGADYVVCDLTQVSVVKNNENGIVIQVNNPLTRD SEQ ID NO.38, derived from brewer's yeast ( Saccharomyces cerevisiae DPP1, amino acid sequence Column MNRVSFIKTPFNIGAKWRLEDVFLLIIMILLLNYPVYYQQPFERQFYINDLTISHPYATTERVNNMLFVYSFVVPSLTILIISILADRRHLIFILYTSLLGLSLAWFSTSFFTNFIKNWIGRLRPDFLDRCQPVEGLPLDTLF TAKDVCTTKNHERLLDGFRTTPSGHSSESFAGLGYLYFWLCGQLLTESPLMPLWRKMVAFLPLLGAALIALSRTQDYRHHFVDVILGSMLGYIMAHFPYRRIPPPIDDPLPFKPLMDDSDVTLEEAVTHQRIPDEELHPLSDEGM SEQ ID NO.39, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae DSK2, amino acid sequence Column MSLNIHIKSGQDKWEVNVAPESTVLQFKEAINKANGIPVANQRLIYSGKILKDDQTVESYHIQDGHSVHLVKSQPKPQTGSAAEANNATATGAAAGTGATPNMSSGQSAGFNPLADLTSARYAGYLNMPSADMFGPDGGALNNDSNNQDELLRMMENPIFQSQMNEMLSNPQMLDFMIQSNPQLQA MGPQARQMLQSPMFRQMLTNPDMIRQSMQFARMMDPNAGMGSAGGAASAFPAPGGDAPEEGSNTNTTSSSNTGNAGTNAGTNAGANTAANPFASLLNPALNPFANAGNAASTGMPAFDPALLASMFQPPAQASQAEDTRPPEERYEHQLRQLNDMGFFDFDRNVAALRRSGGSVQGALDSLLNGDV SEQ ID NO.40, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae DUT1, amino acid sequence Column MTATSDKVLKIQLRSASATVPTKGSATAAGYDIYASQDITIPAMGQGMVSTDISTVPVGTYGRIAPRSGLAVKNGIQTGAGVVDRDYTGEVKVVLFNHSQRDFAIKKGDRVAQLILEKIVDDAQIVVVDSLEESARGAGGFGSTGN SEQ ID NO.41, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae FAB1, amino acid sequence Column SEQ ID NO.42, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae FBP1, amino acid sequence Column MPTLVNGPRRDSTEGFDTDIITLPRFIIEHQKQFKNATGDFTLVLNALQFAFKFVSHTIRRAELVNLVGLAGASNFTGDQQKKLDVLGDEIFINAMRASGIIKVLVSEEQEDLIVFPTNTGSYAVCCDPIDGSSNLDAGVSVGTIASIFRLLPDSSGTINDVLRCGKEMVAACYAMYGSSTHLVLTLGDGVDGFTLDTNLGEFILTHPNLRIPPQKAIYSINEGNTLYWNETIRTFIEKVKQPQADNNNKPFSARYVGSMVADVHRTFLYGGLFAYPCDKKSPNGKLRLLYEAFPMAFLMEQAGGKAVNDRGERILDLVPSHIHDKSSIWLGSSGEIDKFLDHIGKSQ SEQ ID NO.43, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae FBP26, amino acid sequence Column MGYSTISNDNDIKVCVIMVGLPARGKSFISQKIIRYLSWLSIKAKCFNVGNYRRDVSGNVPMDAEFFNFENTDNFKLRELAAQNAIKDIVNFFTKEDGSVAVFDATNSTRKRKWLKDICEKNNIQPMFLESWSNDHELIINNAKDIGSTPDYENFEPHVAEADFLERIRQYERFYEPLDPQKDKDMTFIKLVNIIEEVVINKIRTYLESRIVFYVMNIRPKPKYIWLSRHGESIYNVEKKIGGDSLSERGFQYAKKLEQLVKESAGEINLTVWTSTLKRTQQTANYLPYKKLQWKALDELDAGVCDGMTYEEIEKEYPEDFKARDNDKYEYRYRGGESYRDVVIRLEPVIMELERQENVLIITHQAVLRCIYAYFMNVPQEESPWMSIPLHTLIKLEPRAYGTKVTKIKANIPAVSTYKEKGTSQVGELSQSSTKLHQLLNDSPLEDKF SEQ ID NO.44, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae FCP1, amino acid sequence ColumnMTTQIRSPQGLPYPIQIDKLIPSVGSYLHEGDRLLVYKFWYLVERASDTGDDDNEHDVSPGGSAGSNGVSPPTKQLRESIEFFESPYEGDLISWNVDVGDEVATANQVICEIKRPCNHDIVYGGLCTQCGKEVSADAFDGVPLDVVGDVDLQISETEAIRTGKALKEHLRRDKKLILVVDLDQTIIHCGVDPTIAEWKNDPNNPNFETLRDVKSFTLDEELVLPLMYMNDDGSMLRPPPVRKCWYYVKVRPGLKEFFAKVAPLFEMHIYTMATRAYALQIAKIVDPTGELFGDRILSRDENGSLTTKSLAKLFPTDQSMVVVIDDRGDVWNWCPNLIKVVPYNFFVGVGDINSNFLPKQSTGMLQL GRKTRQKSQESQELLTDIMDNEKKLQEKIDKEVKRQEEKLNHQLATAEEPPANESKEELTKKLEYSASLEVQQQNRPLAKLQKHLHDQKLLVDDDELYYLMGTLSNIHKTYYDMLSQQNEPEPNLMEIIPSLKQKVFQNCYFVFSGLIPLGTDIQRSDIVIWTSTFGATSTPDIDYLTTHLI TKNPSTYKARLAKFNPQIKIVHPDWIFECLVNWKKVDEKPYTLIVDSPISDEELQNFQTQLQKRQEYLEETQEQQHMLTSQENLNLFAAGTSWLNNDDDEPIDTASDDDDDDHDDESDDENNSEGIDRKRSIEDNHDDTSQKKTKAEPSQDGPVQHKGEGDDNEDSDSQLEEELMDMLDD SEQ ID NO.45, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae FIG4, amino acid sequence ColumnMNNDAMEHTLGGGILTTSGSKQRKTSKFVMGKYTLYETKDRMYIVGSNKRETMFRILEIDLTVPRGELTVLEDNVFFTRNEIMNVLASLEEATEDGLHKKITGYGLLGFIKFTCWYYLIMVTKYSQVAVIGGHGIYHIDGIDIIPITPITNNYKKPEKSSDEARLLNIFKDLDTKTFYFSYTITNTLQTNILREKLKAVDRCDITIPCGITDYNEMFVWNNNNLLSPIFACIDTVFDWFQCIIHGFIDQVNVSVLGKSIYITLIARRSHHFAGARFLKRGVNNKGHVANEVETEQIVTDMILTPFHQPGNGFFDSDRYTSFVQHRGSIPLYWTQDASNLTTKPPIRINVVDPFFSPAALHFDNLFQRYGGGTIQILNLIKTKEKTPRETKLLWEFEQCIDYLNEFLPTLKKLDYTSWDMSRASKQDGQGVIEFLEKYAVN TVTTTGIFHNGPDFASTKIQEGICRSNCIDCLDRTNAAQFVIGKRALGCQLKSLGIIDNSYLEYDSDIVNILTELFHDLGDTIALQYGGSHLVNTMETYRKINQWSSHSRDMIESIKRFYSNSFVDAQRQDAINLFLGHYSWREGFPSLWEMNTDFYLHNAYSLNMPKRSYIHWWNDYNIKSVKELINEELIATGNDVTREKIIKNVRGYPGAFDNYWNEYYLPRSVTWIRDLFAYNMNSTRRYHNALSKQDKAMSPFTSRKQSWLNNKLKMITSSKSLEKAEGRVVETTDLDRDTSPKQELELYEHYLHIISDRSQKLEEKMNSFSYSKYPIFISHESSEIPPMRKVIGEPLVDIAEDFTDVYDDDDDDGDDENDEMTTEALLIAPDHVSVDEKFYEKVLNVDDYKPALDDYSAVIHIKPDNLQLYRDLCFSKDIQLDFQ SEQ ID NO.46, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae FIN1, amino acid sequence ColumnMSNKSNRRSLRDIGNTIGRNNIPSDKDNVFVRLSMSPSRTTSQKEFLKPPMRMSPNKTDGMKHSIQVTPRRIMSPECLKGYVSKETQSLDRPQFKNSNKNVKIQNSDHITNIIFPTSPTKLTFSNENKIGDGSLTRIRARFKNGLMSPERIQQQQQQHILPSDAKSNADLCSNTELKDAPFENDLPRAKLKGKNLLVELKKEEEEDVGNGIESLTKSNTKLNSMLANEGKIHKASFQKSVKFKLPDNIVTEETVELKEIKDLLLQMLRRQREIESRLSNIELQLTEISKHK SEQ ID NO.47, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae FPY1, amino acid sequence Column MVKVTAACIIIGDEVLNGKVVDTNSTFFAKYCFDHGIQLKEIATIGDDETQIVDTVRRLVKNYDFIISTGGIGPTHDDITYECMAKSFNLPCELDEECKERMRHKSDPEARLDADALKAHYQMATMPKGTNVKNYYV CDDLWVPICSISHKMYILPGIPQLFARMLKAFTPTLKKIYNLDKDPREYVRYFVRTHLTESQISKELKLIQDESTKVSEAIKIGSYPHFGMGFNTVSILGEKKDDSYLKSIVNRVVNNLEGEVISSELENKFSNQES SEQ ID NO.48, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GAC1, amino acid sequence ColumnMVIQTATTLSPAKARSFPHNDLIKSMSDSLISRPTHPPIRKLKSSLKISHPEPISRSKSEIFCTSPEKNVRFAIELTTVKRFDKNAEPSSISNENSPTLSPVDSNTAADDVQLFNNEDCWFNDSSLVTNLLKNEKKFRYMNSLNNMFKLDLYDSEDEDDIDEHINSQAEYGYTYNSLSTRGKTSENKSATSSLATQATNICDWKLHCTDLVPFKIAPPLFTKTLSASDLQGQLTKYLNGQNVKLHSLTQLGDDSSSKITGLVYVKNLSFEKYLEIKFTFNSWRDIHYVTANFNRTINSNVDEFKFTIDLNSLKYILLIKRIITMEKNTSSCPLNIELCCRYDVNNETYYDNNNGKNYHLFMTTFKKGGETKEKIPVVVEPASQTDAAMSPKEMKAR FVSSNPTLSRFLPQSRKFSEDTDYYNTSPLKHLYHNDTTSWVKPKRLNVVLDKLENATPPPSSALANDTARTGKITKDKNNVLAPPTASNSIDLPILGSQHQSLYGSSSSYSSSSSSISSSLSFASSNNSSTNSSSASCSFPLTELDNFDYANLYEPNDTFTTANLFNHSLNSLMPEISTPSFFGGFRNENTINNND SKNLVTSLEDSYEDKQSVITDTTMDENNKTSTINNSTDTLIKPSKENGTVKENKSSANSTSAPSSSQNRASTILNDHSNGKSDLKYVNYQSLLDSHCFYNHPSPSPNLQSTSFSSAAPFSGISQASDIFDYEDENSDSNQIAGEIDNNSFPPHFYLDEDDKSACLSDDALIDHHRNTNPFINTFSSSPPILSQEVDRWRL SEQ ID NO.49, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GDA1, amino acid sequence ColumnMAPIFRNYRFAIGAFAVIMLILLIKTSSIGPPSIARTVTPNASIPKTPEDISILPVNDEPGYLQDSKTEQNYPELADAVKSQTSQTCSEEHKYVIMIDAGSTGSRVHIYKFDVCTSPPTLLDEKFDMLEPGLSSFDTDSVGAANSLDPLKVAMNYVPIKARSCTPVAVKATAGLRLGDAKSSKILSAVRDHLEKDYPFPVVEGDGVSIMGDEEGVFAWITTNYLLGNIGANGPKLPTAAVFDLGGGSTQIVFEPTFPINEKMVDGEHKFDLKFGDENYTLYQFSHLGYGLKEGRNKVNSVLVENALKDGKILKGDNTKTHQLSSPCLPPKVNATNEKVTLESKETYTIDFIGPDEPSGAQCRFLTDEILNKDAQCQSPPCSFNGVHQPSLVRTFKESNDIYIFSYFYDRTRPLGMPLSFTLNELNDLARIVCKGEETWNSVFSGIAGSLDELESDSHFCLDSFQVSLLHTGYDIPLQRELRTGKKIANKEIGWCLGASLPLLKADNWKCKIQSA SEQ ID NO.50, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GEP4, amino acid sequence Column MNISGTLNTLRLLYNPSLCKPSLVVPTFNDLPIPIHDSIKAVVLDKDNCIAFPHDDKIWPDYLQHWETLRSKYSNKALLIVSNTAGSNSDKDYSQAKLLEDKTGIPVLRHSTKKPGCHNEILDYFYRNKTITNPKEVAVVGDRLFTDILMANLMGSYGVWIRDGVKVSANPLSKFEKKLYNFLGF SEQ ID NO.51, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GIP1, amino acid sequence ColumnMETILQPKARPFESLKRKRFREWLRPSTAHGSLLHSDTLDLRDFAKPNPADTFSNLDSGHCPLVTTPIKYECPDGKSSFFRGDTKFETLFSNRKFYEFKDNLKRGLKKIRHGRNGHQSEKRCPVVEETKKSVSDNLDKPDNNTPCFDRFHTNSKEFETQFDHSNRSQNSEKAYLDNESCWNLSEKFIPFNNLKYEDLKHFEENLQSLAPATFTPIESNESLDRSDSTRGTKRSIRNDSSDTTSEKRLCLKQYSDEPESDHSMESTPSIYITKEVQERIEALSSTDSFLIEKVDFPSNKIGSSASDYESDNEYRNMDEDSINDVTTEKEGNVVIPDSNTSTVDAMEKPIEVSSALKDDTLDKDIDDASSSYSDDVETTFEPVESEELSDLSDTSSSGSSKIYTIPTFRGLTNRTNISQILSKVGKADLSQDNLTHLIKSHQKKKRCVNFRNKRFYDAFNPYVDNEEDAELSDSENISEMDTDLCIKDRSTSSVRFDENSRLLIYKKSKKLNKDETQSGYSTTEMRSILKTKMNSQHDEESQRASKCDTVGVAQFLHYFQYTEYKRQRNEAENYRLRGEQLSKYYSEEYPLDFAAVECEDSVNDKSDIILSMRATERNIGRQLKGISSQGAQIISLDEDVF SEQ ID NO.52, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GIP2, amino acid sequence ColumnMYIKAEQKPQQFERKNEKLDRKKNQQLPDLETDFKGYRVNSDLYNKERDGSTEETLNSLKFLHKPQRVTQMRANRFPEEVQRNTDLNKRIFSAGNDENVDNESCWSKIAAAKNHTSVESLNGSTRPPFKIELPPLSPKSTVPKSFQAEYPEAKSPGNDMNFEYDEEILIPFAPPVYKKSGELLKSSLKRRSKSLPTTPGIRSGNGVQARDGSPMLIRSKSVHFDQAAPVKYFAEDESPINVNKTEQHDNCLSFKHKPVNLMVDPEEETKMLSS GLETTSIDDDLTTVAPKGFAHPAKISSPNNGKGTNNTKLRKSKRFQNLLKNRTDMPPSKSNKKFVNGGGAHEISDRNSKNYHVVGLYSKNFPILSNKNPKSLKLNIFINLSQNKKVFLQELSLYIHRDNNYFSNSSSFNNIPNSHNGNDCNGVAKGYNAGCTRLIAGRILVKNIFYDKRVVVRYTWDSWRTTHEVECVYISDGDGILPGTNMDIFHFIIDDASKVDPRGKLEFCIHYSTRNDYEREEYWDNNNGNNYKVDVVMDGFNDPFAAAA SEQ ID NO. 53, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae GIP3, amino acid sequence Column SEQ ID NO. 54, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae GIP4, amino acid sequence [[ID=5 MVDVQKRKKLLAKAAASASIPAIKGSVPLDSYDIKIIQYKNALYKLNELNRLLNVLVPHLKKKRDNDESYKIIPLVNFILSLCEGPIFNVSPVLAKRYHLLCRFQLIKLSEVQQRLSTNFIDVEGWMFPEEVPLDHYKSCIYNNSLQWKILNSLSCIAQNAIKIYNAKLRQILLERDAYKARSLPFDTSI IEDLLNPVEMTLILDLAVLINDPVRDKSTHSFYKLQWQVMEKNLSCVHSKIFPILRTYYNQLQKFSETRPTSLSNLQKDLPHWEWTHLRIYTFHLRVFSVLCVIISFSRQIFLPNKQHFLDIKTRLSSENVYHYDLIICELMALLSPECDDVTALFELQENLKFWTQTARTDNNSSRTPIFHLQPGLVVE LFNNHICKIIPKLRSIMGLLSNWMDCWKYIEKNYKTFDETNDLRENLKEKLERDKALYLEVKNAKSKLKKKPSITKLPASSSPSPTSSASPSRQASLESIRTRARAHLASNSSRSPSVSPVRTTFNNKNAETKKSVVSPEKRKLINGRRPRSSSLQSYTNKQQTSYLNSTRHPSIAPPSKLNNQRSNSLQSSTMTLNQKIVQDTVRHLMNKSASTPNPSASSSLAPSPKVSSINNTSSGKSSSTLIANSDTLAIETLLDPESNSSELSIKRVRFAGVPPMTEAENPKPTKVGWYKKPAVLHYPPIPASAMIKPLQHKSKYNTLRQEEGFTFRKSLRDGLEWENGESGSETTMMPFGIEIKESTGHRIASKIRSKLR SEQ ID NO. 55, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ GLC7, amino acid sequence ​MDSQPVDVDNIIDRLLEVRGSKPGQQVDLEENEIRYLCSKARSIFIKQPILLELEAPIKVCCTSQSRDRMPRASGVLCVLLLLRQDSYSSYFRLTERIYQASDVSLLICRDSRDNVCVLFIFLSYAFFYQDFLMFCIIIRPAPPIFRKIPLAHTKKRRHFFVMKKTNLFFFAREKGRTKTKKCGFAEGIGRNEAFLTAVIVLLKSTEARFSDKFIYLIFFYFFFFICGDIHGQYYDLLRLFEYGGFPPESNYLFLGDYVDRGKQSLETICLLLAYKIKYPENFFILRGNHECASINRIYGFYDECKRRYNIKLWKTFTDCFNCLPIAAIIDEKIFCMHGGLSPDLNSMEQIRRVMRPTDIPDVGLLCDLLWSDPDKDIVGWSENDRGVSFTFGPDVVNRFLQKQDMELICRAHQVVEDGYEFFSKRQLVTLFSAPNYCGEFDNAGAMMSVDESLLCSFQILKPAQKSLPRQAGGRKKK SEQ ID NO. 56, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ GLC8, amino acid sequence ​ MGGILKNPLALSPEQLAQQDPETLEEFRRQVYENTQKNAKLTSHKRNIPGLDNTKEEGEIIGTSSTFLPKDTLSLKHEQDMLAKMTPEERVQWNQRNLAENEITKKQFQDIHIDEPKTPYQGAVDPHGEYYRVDDDEDEDNSDKKPCQVANDDIDDLSLGEPEFEIKENKQPDFETNDENDEDSPEARHKKFEEMRKKHYDVRAIFNKKSREALKDEDEDEDDSTTKEP SEQ ID NO. 57, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ GLN3, amino acid sequence ​MQDDPENSKLYDLLNSHLDVHGRSNEEPRQTGDSRSQSSGNTGENEDIAFASGLNGGTFDSMLEALPDLYFTDFVSPFTAAATTSVTTKTVKDTTPATNHMDDDIAMFDSLATTQPIDIAASNQQNGEIAQLWDFNVDQFNMTPSNSSGSATISAPNSFTSDIPQYNHGSLGNSVSKSSL FSYNSSTSNSNINQPSINNNSNTNAQSHHSFNIYKLQNNNSSSAMNITNNNNSNNSNIQHPFLKKSDSIGLSSSNTTNSVRKNSLIKPMSSTSLANFKRAASVSSSISNMEPSGQNKKPLIQCFNCKTFKTPLWRRSPEGNTLCNACGLFQKLHGTMRPLSLKSDVIKRISKKRAKQDTSN IAQNTPNASATSSTSVTTTNAKPIRSRKKSLQQNSLSRVIPEEIISDNIGNTNNILNVNRGGYNFNSVPSPVLMNSQSYNSNANFNGASNANLNSNNLMRHNSNTVTPNFRRSSRRSSTSSNTSSSKSSSRSVVPILPKPSPNSANSQQFNMNLMNTTNNISAGNSVASSPRIISSANFNSNSPLQQNLLSNSFQRQGMNIPRRKMSRNASYSSSFMAASLQQLHEQQVDVNSNTNTNSNRQNWNSSNSVSTNSRSSNFVSQKPNFDIFNTPIDSPSVSRPSSRKSHTSLLSQQLQNSESNSFISNHKFNNRLSSDSTSPIKYEADVSTGGKISEDNSTKGSSSKESSAIADELDWLKFGI SEQ ID NO. 58, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ GPP1, amino acid sequence ​MPLTTKPLSLKINAALFDVDGTIIISQPAIAAFWRDFGKDKPYFDAEHVIHISHGWRTYDAIAKFAPDFADEEYVNKLEGEIPEKYGEHSIEVPGAVKLCNALPKEKWAVATSGTRDMAKKWFDILKIKRPEYFITANDVKQGKPPHEPYLKGRNGLGFPINEQDPSKSKVVVFEDAPAGIAAGKAAGCKIVGIATTFDLDKEKGCDIIVKNHESRVGEYNAETDEVELIFDDYLYAKDDLLKW SEQ ID NO. 59, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ GPP2, amino acid sequence ​ MGLTTKPLSLKVNAALFDVDGTIIISQPAIAAFWRDFGKDKPYFDAEHVIQVSHGWRTFDAIAKFAPDFANEEYVNKLEAEIPVKYGEKSIEVPGAVKLCNALNALPKEKWAVATSGTRDMAQKWFEHLGIRRPKYFITANDVKQGKPHPEPYLKGRNGLGYPINEQDPSKSKVVVFEDAPAGIAAGKAAGCKIIGIATTFDDLFLKEKGCDIIVKNHESIRVGGYNAETEDEVEFIFDDYLYAKDDLLKW SEQ ID NO. 60, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ GSY2, amino acid sequence ​MSRDLQNHLLFETATEVANRVGGIYSVLKSKAPITVAQYKDHYHLIGPLNKATYQNEVDILDWKKPEAFSDEMRPVQHALQTMESRGVHFVYGRWLIEGAPKVILFDLDSVRGYSNEWKGDLWSLVGIPSPENDFETNDAILLGYTVAWFLGEVAHLDSQHAIVAHFHEWLAGVALPLCRKRRIDVVTIFTTHATLLGRYLCASGSFDFYNCLESVDVDHEAGRFGIYHRYCIERAAAHSADVFTTVSQITAFEAEHLLKRKPDGILPNGLNVIKFQAFHEFQNLHALKKEKINDFVRGHFHGCFDFDLDNTLYFFIAGRYEYKNKGADMFIEALARLNYRLKVSGSKKTVVAFIVMPAKNNSFTVEALKGQAEVRALENTVHEVTTSIGKRIFDHAIRYPHNGLTTELPTDLGELLKSSDKVMLKRRILALRRPEGQLPPIVTHNMVDDANDLILNKIRQVQLFNSPSDRVKMIFHPEFLNANNPILGLDYDEFVRGCHLGVFPSYYEPWGYTPAECTVMGVPSITTNVSGFGSYMEDLIETNQAKDYGIYIVDRRFKAPDESVEQLVDYMEEFVKKTRRQRINQRNRTERLSDLLDWKRMGLEYVKARQLALRRGYPDQFRELVGEELNDSNMDALAGGKKLKVARPLSVPGSPRDLRSNSTVYMTPGDLGTLQEVNNADDYFSLGVNPAADDDDDGPYADDS SEQ ID NO. 61, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ HIS2, amino acid sequence ​MHSHHSHSGDYSAHGTDPLDSVVDQVVNLNFHTYCLTEHIPRIEAKFIYPEEQSLGKNPEEVISKLETSFKNFMSHAQEIKTRYADRPDVRTKFIIGMEIESCDMAHIEYAKRLMKENNDTLKFCVGSVHHVNGIPIDFDQQQWYNSLHSFNDNLKDFLLSYFQSQYEMLINIKPLVVGHFDLYKLFLPNDMLVNQKSGNCNEETGVPVASLDVISEWPEIYDAVVRNLQFIDSYGGAIEINTSALRKGLEEPYPSKTLCNLVKKHCGSRFVLSDDAHGVAQVGVCYDKVKKYIVDVLQLEYICYLEESQSPENVLTVKRLPISQFVNDPFWANI SEQ ID NO. 62, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ HNT2, amino acid sequence ​ MPESQDYFKTLQLIHRFIKWQYKADSINVAIQDGPEAGQSVPHLHTHIIPRYKINNVGDLIYDKLDHWDGNGTLTDWQGRRDEYLGVGGRQARKNNSTSATVDGDELSQGPNVLKPDSQRKVRALTEMKKEAEDLQARLEEFVSSDPGLTQWL SEQ ID NO. 63, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ HRR25, amino acid sequence ​MDLRVGRKFRIGRKIGSGSFGDIYHGTNLISGEEVAIKLESIRSRHPQLDYESRVYRYLSGGVGIPFIRWFGREGEYNAMVIDLLGPSLEDLFNYCHRRFSFKTVIMLALQMFCRIQYIHGRSFIHRDIKPDNFLMGVGRRGSTVHVIDFGLSKKYRDFNTHRHIPYRENKSLTGTARYASVNTHLGIEQSRRDDLESLGYVLIYFCKGSLPWQGLKATTKKQKYDRIMEKKLNVSVETLCSGLPLEFQEYMAYCKNLKFDEKPDYLFLARLFKDLSIKLEYHNDHLFDWTMLRYTKAMVEKQRDLLIEKGDLNANSNAASASNSTDNKSETFNKIKLLAMKKFPTHFHYYKNEDKHNPSPEEIKQQTILNNNAASSLPEELLNALDKGMENLRQQQPQQQVQSSQPQPQPQQLQQQPNGQRPNYYPEPLLQQQQRDSQEQQQQVPMATTRATQYPPQINSNNFNTNQASVPPQMRSNPQQPPQDKPAGQSIWL SEQ ID NO. 64, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ ICE2, amino acid sequence ​MTSLSKSFMQSGRICAACFYLLFTLLSIPISFKVGGLECGLSFTVTLFTLYFITTTLNVLARRHGGRLYIFFTNCLYYSQHFIIASLLYLFLSGFSNDELGNVLKNKYNESESFLEALKNSLNSNQINYVLYYYYYRFVVQPWQFVLTKSTPFFTLSEGFFTILAIQAVGETNRWLSNDLNSNTWIISSLLTSGGVITASLYYLYRIYVTPIWPLSIQTASLLGLVLSMVCGLGLYGIVSQKGSVIESSLFFAYIVRCIYEISPKLATTATDEILNLFKDVWQKHQRNLPTADNLLCYFHNVILKNAEVLWGSFIPRGRKKTGDFHDKLISILSFEKVSLISKPFWKFFKNFTFSVPLSINEFCQVTIKMASESVSPAIVINLCFRVLMFYSATRIIPALQRKNDKQLRKSRRIMKGLYWYSPCILIAMYTHLILQYSGELKKDLCIWGCSEKWFGVDQPEIIVDSWGFWNWCNIFCTILVYATELIGSGS SEQ ID NO. 65, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ IGO1, amino acid sequence ​ MSNENLSPNSSNPDLTKLNNGESGTIDTSKFSPNEMKLYKMYGKLPSKKDIFKHTMQKRKYFDSGDYALQKAGIQNNDPINYGKNNLPLTNPSKLREDIIKRRISTCPSTASTAGVVDNATLIQKEGSISSGPPSSNNGTIGGGSTSSTPVGNHSSSSSSLYTESPIR SEQ ID NO. 66, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). ​ IGO2, amino acid sequence ​ MSEDLSPTSSRVDLSNPHGFTKEGVDLSKLSPQELKLYKMYGKLPSKKDLLRHKMQDRQYFDSGDYALKKAGVIKSDDVIVNNSSNNLPVTNPSGLRESIIRRRMSSSSGGDSISRQGSISSGPPPRSPNK SEQ ID NO. 67, derived from Saccharomyces cerevisiae (Saccharomyces cer ​ INM1, amino acid sequence ​MTIDLASIEKFLCELATEKVGPIIKSKSGTQKDYDLKTGSRSVDIVTAIDKQVEKLIWESVKTQYPTFKFIGEESYVKGETVITDDPTFIIDPIDGTTNFVHDFPSFSCTSGLTVNKEPVVGVIYNPHINLLVSASKGNGMRVNNKDYDYKSKLESMGSLILNKSVVALQPGSAREGKNFQTKMATYEKLLSCDYGFVHGFRNLGSSAMTMAYIAMGYLDSYWDGGCYSWDVCAGWCILKEVGGRVVGANPGEWSIDVDNRTYLAVRGTINNESDEQTKYITDFWNCVDGHLKYD SEQ ID NO. 68, derived from brewer's yeast ( ​ INM2, amino acid sequence ​ MVLTRQVLEEVENTFIELLRSKIGPLVKSHAGTNFCSYDDKANGVDLVTALDKQIESIIKENLTAKYPSFKFIGEETYVKGVTKITNGPTFIVDPIDGTTNFIHGYPYSCTLGLAEMKGKPVVGVVFNPHNLNQLFHASKGNGAFLN DQEIKVSKRPLILQKSLIALEGGSERTEGSQGNFDKKMNTYKNLLSESGAFVHGFRSAGSAAMNICYVASGMLDAYWEGGCWAWDVCAGWCILEEAGGIMVGGNCGEWNIPLDRRCYLAIRGGCESMEQKRFAESFWPHVAGELEY SEQ ID NO. 69, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae INP51, amino acid sequence ColumnMRLFIGRRRSRSIVISSNNYCLSFQRLRSIPGASSQQRQLSKTPSVTIKSYPDTDLSSDSNYLEVKSCIFNGLLGLVCLNGDIYVAVISGVQNVGFPRWKLIDHQVRPSESIYKVLDVDFYSLENDVFDYLLCERSEQNYDKLIHEHPCGPLKKLFSDGTFYYSRDFDISNIVKNHGLSHNLEYTVDNQDLSFIWNANLASEVINWRSKISNEEKQLFANAGFLTFVIRGYCKTALI EDGPNTASITIISRISTESKQDTLELEGISEDGRVSLFVETEIVVTTEKFIFSYTQVNGSIPLFWESVESQLLYGKKIKVTKDSIEAQGAFDRHFDNLTSKYGVVSIVNIIKPKSESQEKLALTYKDCAESKGIKITNIEYSSSVLTKSPHKLLYLLKQDIYEFGAFAYDISRGIYFAKQTGVLRISAFDSIEKPNTVERLVSKEVLELTTNEIDVFELTSPFLDAHDKLWSENYW LDRTYTKHTKNSGKYTKVYSKLFGSRVRLYDPLHIYISQYLKQLRSKYTFEKDISIFAGTFNISGKIPKDDIKDWIFPKSMSKEDEMADLYVIGLEEVVELTPGHMLATDPYVRQFWEKKILTLNGPGRKKKYIRLWSTQLGGILLLLFMNETEYSKVKHIEGDVKKTGFGGMASNKGAVAVSFKYSATRFCVLVSHLAAGLENVEQRHNDYKTIAKSIRFSKGLRIKDHDAIIWMGDFNYRILMSNEDVRRKIVSKEYASLFEKDQLNQQMIAGESFPYFHEMAIDFPPTYKFDPGTKNYDTSEKMRIPAWTDRILSRGEVLEQLEYKCCEDILFSDHRPVYAIFRARVTVVDEQKKTTLGTQIYEKIMERLEGLDDDEKIAVLSDDAFVIESFEGSDSIAGPTHSPTIPEPKRGRKPPPPSSDLKKWWIGSGKQVKVVLDVDPAVYMINPKRDPNPFVENEDEPLFIER SEQ ID NO.70, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae INP52, amino acid sequence Column SEQ ID NO.71, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae INP53, amino acid sequence Column SEQ ID NO.72, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae INP54, amino acid sequence Column MNKTNWKVSVTTFNCGKEFPVENSKAIVKQLLFPYDDGISQLELQDLYVLGFQEVVPIWQGSFPAVNRDLIDRITTTAVNCLNEKVSATQGDEQYSCLGVNSLGAITIIVLYNNNALKVKDDILKRNGKCGWFGTHLKGGTLISFQMTRNGEENWERFSYICAHLNANEGVNNRNQRIDDYKRIMSEVCDSEVAKSDHFFLGDLNFRVTSTYDPTTNYSSTTTLRRLLENHEELNLLRKGEDEPLCKGFQELKITFPPTYKFKLFEKETYNTKRIPSWCDRILYKSYAVPTFAQEGTYHSVPRSNALLFSDHQPVNLTVRLPRSTGTPVPLSLHIEKYPLSWSSGLIGQIGDAVIGYCGWLVTKNVHYWILGSLLLYLLLKIL SEQ ID NO.73, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae IPP1, amino acid sequence Column MTYTTRQIGAKNTLEYKVYIEKDGKPVSAFHDIPLYADKENNIFNMVVEIPRWTNAKLEITKEETLNPIIQDTKKKGKLRFVRNCFPHHGYIHNYGAFPQTWEDPNVSHPETKAVGDNDPIDVLEIGETIAYTGQVKQVKALGI MALLDEGETDWKVIAIDINDPLAPKLNDIEDVEKYFPGLRATNEWFRIYKIPDGKPENQFAFSGEAKNKKYALDIIKETHDSWKQLIAGKSSDSKGIDLTNVTLPDTPTYSKAASDAIPPASPKADAPIDKSIDKWFFISGSV SEQ ID NO.74, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae ISN1, amino acid sequence ColumnMSSRYRVEYHLKSHRKDEFIDWVKGLLASPFVLHAVSHEGDYNDDLATTQRVRSQYADIFKDIEGLIKDKIEFDSRNMSQDEIEDGASSQSLNILGQSRLNLLVPSIGTFFTELPLEQAFLWEDSQRAISARRMVAPSFNDIRHILNTAQIFHFKKQENLHNGKVLRLVTFDGDVTLYEDGGSLVYTNPVIPYILKLLRCGINVGIVTAAGYDEAGTYENRLKGLIVALHDSTDIPVSQKQNLTIMGGESSYLFRYYEDPEEDNFGFRQIDKEEWLLPRMKAWSLEDVEKTLDFAERTLNRLRKRLNLPSEISIIRKVRAVGIVPGERYDEASKRQVPVKLDREQLEEIVLTLQNTLESFAPSRRIQFSCFDGGSDVWCDIGGKDLGVRSLQQFYNPESPIQPSETLHVGDQFAPVGSANDFKARLAGCTLWIASPQETVNYLHRLLETD SEQ ID NO.75, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae KAE1, amino acid sequence Column MVNLNTIPPKNGRDYYIALGLEGSANKLGVGIVKHPLLPKHANSDLSYDCEAEMLSNIRDTYVTPPGEGFLPRDTARHHRNWCIRLIKQALAEADIKSPTLDIDVICFTKGPGMGAPLHSVVIAARTCSLLWDVPLVGVNHCIGHIEMGREITKAQNPVVLYVSGGNTQVIAYSEKRYRIFGETLDIAIGNCLDRFARTLKIPNEPSPGYNIEQLAKKAPHKENLVELPYTVKGMDLSMSGILASIDLLAKDLFKGNKKNKILFDKTTGEQKVTVEDLCYSLQENLFAMLVEITERAMAHVNSNQVLIVGGVGCNVRLQEMMAQMCKDRANGQVHATDNRFCIDNGVMIAQAGLLEYRMGGIVKDFSETVVTQKFRTDEVYAAWRD SEQ ID NO.76, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae LCB3, amino acid sequence ColumnMVDGLNTSNIRKRARTLSNPNDFQEPNYLLDPGNHPSDHFRTRMSKFRFNIREKLLVFTNNQSFTLSRWQKKYRSAFNDLYFTYTSLMGSHTFYVLCLPMPVWFGYFETTKDMVYILGYSIYLSGFFKDYWCLPRPRAPPLHRITLSEYTTKEYGAPSSHTANATGVSLLFLYNIWRMQESSVMVQLLLSCVVLFYYMTLVFGRIYCGMHGILDLVSGGLIGIVCFVVRMYFKYRFPGLRIEEHWWFPLFSVGWGLLLLFKHVKPVDECPCFQDSVAFMGVVSGIECCDWLGKVFGITLVYNLKPNCGWRLTLARLLVGLPCVVIWKYVISKPMIYTLLIKVFHLKDDRNVAARKRLEATHKEGASKYECPLYIGEPKIDILGRFIIYAGVPFTVVMCSPVLFSLLNIA SEQ ID NO.77, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae LPP1, amino acid sequence Column MISVMADEKHKEYFKLYYFQYMIIGLCTILFLYSEISLVPRGQNIEFSLDDPSISKRYVPNELVGPLECLILSVGLSNMVVFWTCMFDKDLLKKNRVKRLRERPDGISNDFHFMHTSILCLMLIISINAALTGALKLIIGNLRPDFVDRCIPDLQKMSDSDSLVFGLDICKQTNKWILYEGLKSTPSGHSSFIVSTMGFTYLWQRVFTTRNTRSCIWCPLLALVVMVSRVIDHRHHWYDVVSGAVLAFLVIYCCWKWTFTNLAKRDILPSPVSV SEQ ID NO.78, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae LSB6, amino acid sequence ColumnMSNEAYQHDHTVNPHQKIVVNSYDWLQFRDEQDHCKSKNPITHASPGVGSNAQNSDIAEAPQVFHPSYQSLVNVPSESPRPDQTSGSNPAVGLLHNAEDKASGQEEEGSQYEIQYSVFRPLHAYPTKGLAYEQLRRKEEQEQRENFNHLVSDCIEAVETFGRELERIQTGSSGSYFVYGTRADESVPVGVFKPKDEEPYGPFSPKWTKWAHRTFFPCLFGRSCLIPNLGYICESAASLLDRRLETHLVPYTDTASIESFNFYDNRKKWVLGYNLQKKKQKKLGSFQLFLKEYINADEFFHKYPLPGMYSDVKHSFHRKSSGEDINHKPETTRNLTDETEPSKQINSSPISTESEENSKFEWTESSLSQFRLELEKLIILDYIMRNTDRGLDNWMVKLIKLSNNKWRLKLAAIDNGLSFPWKHPDEWRLYPYGWLYLPLQLLAKPFSEQMRSHFLPILTSTNWWEESYQEFLALFSRDQDFNVRMWKKQWAVLKGQAFNVVETLKDPRQGPLELVRRTRCQVIDEKMQVPCCPPPVSIFKNAIDEPIGSYSTSPMVLPSTPSTIPFHAHNQSNSNPVYYDSTLHPFANKTVIAERLQIVNSTPVFTWC SEQ ID NO.79, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae LTE1, amino acid sequence Column SEQ ID NO.80, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae LTP1, amino acid sequence Column MTIEKPKISVAFICLGNFCRSPMAEIFKHEVEKANLENRFNKIDSFGTSNYHVGESPDHRTVSICKQHGVKINHKGKQIKTKHFDEYDYIIGMDESNINNNLKKIQPEGSKAKVCLFGDWNTNDGTVQTIIEDPWYGDIQDFEYNFKQITYFSKQFLKKEL SEQ ID NO.81, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae MET22, amino acid sequence Column MALERELLVATQAVRKASLLTKRIQSEVISHKDSTTITKSDNSPVTTGDYAAQTIIINAIKSNFPDDKVVGEESSSGLSDAFVSGILNEIKANDEVYNKNYKKDDFLFTNDQFPLKSLEDVRQIIDFGNYEGGRKGRFWCLDPIDGTKGFLRGEQFAVCLALIVDGVVQLGCIGCPNLVLSSYGAQDLKGHESFGYIFRAVRGLGAFYSPSSDAESWTKIHVRHLKDTKDMITLEGVEKGHSSHDEQTAIKNKLNISKSLHLDSQAKYCLLALGLADVYLRLPIKLSYQEKIWDHAAGNVIVHEAGGIHTDAMEDVPLDFGNGRTLATKGVIASSGPRELHDLVVSTSCDVIQSRNA SEQ ID NO.82, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae MIG1, amino acid sequence ColumnMQSPYPMTQVSNVDDGSLLKESKSKSKVAAKSEAPRPHACPICHRAFHRLEHQTRHMRIHTGEKPHACDFPGCVKRFSRSDELTRHRRIHTNSHPRGKRGRKKKVVGSPINSASSSATSIPDLNTANFSPPLPQQHLSPLIPIAIAPKENSSRSSTRKGRKTKFEIGESGGNDPYMVSSPKTMAKIPVSVKPPPSLALNNMNYQTSSASTALSSLSNSHSGSRLKLNALSSLQMMTPIASSAPRTVFIDGPEQKQLQQQQNSLSPRYSNTVILPRPRSLTDFQGLNNANPNNNGSLRAQTQSSVQLKRPSSVLSLNDLLVGQRNTNESDSDFTTGGEDEEDGLKDPSNSSIDNLEQDYLQEQSRKKSKTSTPTTMLSRSTSGTNLHTLGYVMNQNHLHFSSSSPDFQKELNNRLLNVQQQQQEQHTLLQSQNTSNQSQNQNQNQMMASSSSLSTTPLLLSPRVNMINTAISTQQTPISQSDSQVQELETLPPIRSLPLPFPHMD SEQ ID NO.83, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae MIH1, amino acid sequence ColumnMNNIFHGTEDECANEDVLSFQKISLKSPFGKKNIFRNVQTFFFKSKHSNVDDDLINKENLAFDKSPLLTNHRSKEIDGPSPNIKQLGHRDELDENENENDDIVLSMHFASQTLQSPTRNSSRRSLTNNRDNDLLSR IKYPGSPQRSSSFSRSRSLSRKPSMNSSSNSSRRVQRQDGKIPRSSRKSSQKFSNITQNTLNFTSASSSPLAPNSVGVKCFESCLAKTQIPYYYYDRNSNDFFPRISPETLKNILQNNMCESFYNSCRIIDCRFEYEYT GGHIINSVNIHSRDELEYEFIHKVLHSDTSNNNTLPTLLIIHCEFSSHRGPSLASHLRNCDRIINQDHYPKLFYPDILILDGGYKAVDFNFPELCYPRQYVGMNSQENLLNCEQEMDKFRRESKRFATKNNSFRKLASPNFYRDSHQSSTTMASSALSFRFEPPKLSLNHRVSSGSSLNSSESTGDENFFPILSKSSMSSNSNLSTSHMLLMDGLDTPSYFSFEDERGNHQQVSGDEEQDGDFTFVGSDREDLPRPARRSLFPSLETEDKK SEQ ID NO.84, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae MSG5, amino acid sequence ColumnMQFHSDKQHLDSKTDIDFKPNSPRSLQNRNTKNLSLDIAALHPLMEFSSPSQDVPGSVKFPSPTPLNLFMKPKPIVLEKCPPKVSPRPTPPSLMRRSEASIYTLPTSLKNRTVSPSVYTKSSTVSSISKLSSSSPLSSFSEKPHLNRVHSLSVKTKDLKLKGIRGRSQTISGLETSTPISSTREGTLDSTDVNRFSNQKNMQTTLIFPEEDSDLNIDMVHAEIYQRTVYLDGPLLVLPPNLYLYSEPKLEDILSFDLVINVAKEIPNLEFLIPPEMAHKIKYYHIEWTHTSKIVKDLSRLTRIIHTAHSQGKKILVHCQCGVSRSASLIVAYIMRYYGLSLNDAYNKLKGVAKDISPNGLIFQLMEWGTMLSKNSPGEEGETVHMPEEDDIGNNEVSSTTKSYSSASFRSFPMVTNLSSSPNDSSVNSSEVTPRTPATLTGARTALATERGEDDEHCKSLSQPADSLEASVDNESISTAPEQMMFLP SEQ ID NO.85, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae MSS4, amino acid sequence ColumnMSVLRSQPPSVVPLHLTTSTRKTEKEPSSLHLHSAIIERHQDRSVPNSNSNPDSNHRIKKDRNNHTSYHSSSNSNMESPRLSDGESSTPTSIEELNPTINNSRLVKRNYSISIDPLHDNSNNNTDDDHPNTITSPRPNSTSKEMQKYSFPEGKESKKITTPSLNSNNCLDLDNSSLVHTDSYIQDLNDDHILLNKRVSRRSSRISAVTATSTTIKQRRNTQDSNLPNIPFHASKHSQILPMDDSDVIKLANGDTSMKPSATKISHSMTSLPLHPLPQPSQKSKQYHMISKSTTSLPPENDHYYQHSRGTNHNHAANAAAAVNNNTTTTTAATGLKRSESAATAEIKKMRQSLLHKREMKRKRKTFLVDDDRVLIGNKVSEGHVNFIIA YNMLTGIRVAVSRCSGIMKPLTPADFRFTKKLAFDYHGNELTPSSQYAFKFKDYCPEVFRELRALFGLDPADYLVSLTSKYILSELNSPGKSFFYYSRDYKYIIKTIHHSEHIHLRKHIQEYYNHVRDNPNTLICQFYGLHRVKMPISFQNKIHRKIYFLVMNNLFPPHLDIHITYDLKGSTWGRFTNLDKERLAKDRSYRPVMKDLNWLEEGQKIKFGPLKKKTFLTQLKKDVELLAKLNTMDYSLLIGIHDINKAKEDDLQLADTASIEEQPQTQGPIRTGTGTVVRHFFREFEGGIRASDQFNNDVDLIYYVGIIDFLTNYSVMKKLETFWRSLRHDTKLVSAIPPRDYANRFYEFIEDSVDPLPQKKTQSSYRDDPNQKNYKD SEQ ID NO.86, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae NEM1, amino acid sequence ColumnMNALKYFSNHLITTKKQKKINVEVTKNQDLLGPSKEVSNKYTSHSENDCVGEVDQQYDHSSSHLKESDQNQERKNSVPKKPKALRSILKEKIASILWALLLFLPYYLIIKPLMSLWFVFTFPLSVIERRVKHTDKRNRGSNASENELPVSSSNINDSSEKTNKPKNCNLNTIPEAVDDLNASDEIILQRDNVKGSLLRAQSVKSRPRSYSKSELSLSNHSSSSNTVFGTKRMGRFLFPKKLIPKSVLNTQKKKKLVIDLDETLIHSASRSTTHSNSSQGHLVEVKFGLSGIRTLYFIHKRPYCDLFLTKVSKWYDLIIFTASMKEYADPVIDWLESSFPSFSSKRYYRSDCVLRDGVGYIKDLSIVKDSEENGKGNSSSLDDVIIIDNSPVSYAMNVDNAIQVEGWISDPTDTDLLNLLPFLEAMRYSTDVRNILALKHGEKAFNIN SEQ ID NO.87, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae NET1, amino acid sequence Column SEQ ID NO.88, derived from brewer's yeast ( Saccharomyces cerevisiae NPP1, amino acid sequence Column MELQNDLESLDNELNDFSEDPFRDDFITDEDAVRSGWRSAWTRMKYWFYKNRLKWTNNPIVIGDAKDSRDGSNFRRGIPLYELDANGQPIDTELVDENELSFGTGFHSKVPFKIIFRTLFGSLVFAIFLILMINIAKPHHSTRVLSHFGSPEFDPYVKYFNGTHEFFPLTIVISLDGFHPSLISKRNTPFLHDLYELKYDGGMNITSTPFMVPSFPTETFPNHWTLVTGQYPIHHGIVSNVFWDPDLNEEFHPGVLDPRIWNNNDTEPIWQTVQSAFDGDIPFKAATHMWPGSDVNYTKYNEEKLQPEHKNPIARERTPFYFDEFNAKEPLSQKLSKIIEYVDMSTLNERPQLILGYVPNVDAFGHKHGYPSESEYYYEDFTETLGEVDTFLKQLVESLQERNLTSFTNLVIVSDHGMSDIVVPSNVIIWEDLLDEKLRKDYVSHAYLEGPMMAISLKDSGNINEVYHNLKTSIDEDKYTVYVNGNFPKEWNFNDGKNHHMASIWIVPEPGYAVMKKEQLKKVAKGDHKDKNEDNVFTIGSHGYDNNAIDMRSVFIGMGPYFPQGYIEPFQNTEIYNLLCDICGVAEKDRNSNDGTGMLMNQLREPQSSEEVEIEDDFDYLVSKFGEFSTYNIIWGGYPEETEQDNVDNDNDDNDDGNTDEIAAMPSSSLTIKLEMTTSIPSATETLLGETSPSSRSSSSSSIQASATASTVGDWLQDIINDAKDLIDDIIDSIDDLVDSDT SEQ ID NO.89, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae NPP2, amino acid sequence ColumnMLLFEQPVDLEKNNEDDTNIKPFAISRHFLLKLLLCGIILIELLLYSKCPKPIDNGPRTIANRSNTYFNGTHDFKTLTILISIDGFHPRLIDAKYTPFLYNLHNLRSPYDMNITTAPYMIPSFPTQTFPNHWSMVTGKYPIEHGIVSNIFWDNFTSSEFRPNNLDARIWSNTADPIWQLLQTESQGEYKVATHMWPGSEVVYEDHGDVPRERMPFYFGKFNQWEKLQDKLAQIFRYIDMPQLKDRP ELVISYIPNVDSYGHSFGYDLRDKRLQKLIGEVDGFFLDLIEGLQKRNLLKISNVMIVSDHGMSNVNANDGEHVVVWERVFPADAMSAFISHLYNEGPMMMVCLKNPRDKQWICDLIEAQLEKAYGDEISRKFHILVKEDDFDPSWKYFQYDNRKHRYDDRVGDIWILADEYYAIVKEMGDVPIGIMGTHGYNFNNCSDMASIFIGMGPMFNNEVVPPFENIEVYNMLIKASALLGEEKTKKEKSLLQ SEQ ID NO.90, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae NPY1, amino acid sequence Column MSTAVTFFGQHVLNRVSFLRCSKEFIKKSLNHDSTVFIPFIEALISPENGDLVQLSNSVKSYKNILSAIVPLYTTLLNTTRSRSDESGINVTFLGLLEGTDSAFNFEWSNISYKGTPPYFGLDIRVTESTLFKKVDFEPIFSYKPVTRDHIFKQTNEDASLYSQGKMYLDWLAKYKFCPGCGSPLFVEAG TKLQCSNENRNVYCNVRDARINNVCFPRTDPTVIIALTNSDYSKCCLARSKKRYGDFVLYSTIAGFMEPSETIEEEACIREIWEETGISCKNIDIVRSQPWPYPCSLMIGCLGIVQFNSKNEVINLNHDDELLDAQWFDTTEIIQALDKYAGGYRVPFKNDINLPGSTTIAFQLINHVCENYKNLRKTSSSHL SEQ ID NO.91, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae OCA1, amino acid sequence ColumnMTSKVGEYEDVPEDESRLTEENVSVPEEEVEDEDEEEDDDDDHIYINEETESGREKVLVSHAPQERIVPPLNFCPVERYLYRSGQPSPVNFPFLLNLKLKTIIWLSNEEPQDTLLEFCDTHRINLQFAAINPDAGEDDNPWDGLTEHSIINVLQTIVTQENYPLLVCCGMGRHRTGTVIGCLRRIMGWNLASVSEEYRRFTGSRGGRILVELLIEAFDTNLVKIDKNKAPSWLLTALE SEQ ID NO.92, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae OCA2, amino acid sequence Column MKYIPPLNFSPVVSTDVSLYRSGYPMPLNYSFIKHQLHLKTIIYIGDKDRPLEEYQSFLESEKIKYYHIFMDSSRDEGIQERMNQVLHLVLDVRNYPILVHSNKGKHRVGVVVGIIRKLLQGWSTAGICQEYGLFSGGMKDGVDLEFITMFETNLKIPRNVIPGFAKHCLYLNELEAAEGSDDESGSESILTAKQPI SEQ ID NO.93, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae OCA4, amino acid sequence Column MLVPPANFGIAEEGIYRCSKVETLNLSFLETLNLKTAIFIGGQEPSKFFKDFFTRSSIKWIVLRMSDFSAAAVPVKSSSVSNANLYSNNNSTLSLQEEKKKSTANGSQNSTTGDPVIQEELAYHLTDNDDLMLIKSTCLKRTFKTLLNVDNYNVLLVDKTALVIGILRKIQKWNIASIINEYRLFSGKNRNYFAETFLEIINIEIEQEKDNKTIVDNKAKKLPLENNRTHSIEYKANSGKLIRVNEDDLCREPEVPQRLLTLINQIETKVKNNKVLQVSGVLGDDLKKTSSDLGIFGHRYRLAFNKKENGDYGYYKARGKDNVKIRIPCDSELPDWFKFQRDLWEKENVPEEHHFYREHIFT SEQ ID NO.94, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae OCA6, amino acid sequence ColumnMTLVTPLQFSTVQPNLYRGSYPREINLPFLRTLRLKYILSLTPEPLSTDPLMVKFCEENNIKTIHIKCQSERKADKTKPKIKRKKKTVPIEYDVVVRCVKFLIDKGHYPCYMHCTNGELIISLVVACMRKFSYWSTVSILNEFLVYNSSINIHERNFIENFNSEIEVDDLDIKDKVPWITVRYIARTATESKDELRVDDANASEKVARVSSVSNSLPKLKFHSM SEQ ID NO.95, derived from Saccharomyces cerevisiae (Saccharomyces cerevi Saccharomyces cerevisiae PAH1, amino acid sequence ColumnMQYVGRALGSVSKTWSSINPATLSGAIDVIVVEHPDGRLSCSPFHVRFGKFQILKPSQKKVQVFINEKLSNMPMKLSDSGEAYFVFEMGDQVTDVPDELLVSPVMSATSSPQSPETSILEGGTEGEGEGENENKKKEKKVLEEPDFLDINDTGDSGSKNSETTGSLSPTESSTTTPLDSVEERKLEQRTKNFQQKLNKKLKTEIHIPSKLDNNGDLLDTEGYKPNKNMMHDDDIQLKQLLKDEFGNDSDISSFIKEDKNGNIKIVNPYEHLTDLSPPGTTPMATSGSVLGLDAMESGSTLNSLSSSPSGSDTEDETSFSKEQSSKSEKTSKKGTAGSGETEKRYIRTIRLTNDQLKCLNLTYGENDLKFSVDHGKAIVTSKLFVWRWDVPIVISDIDGTITKSDALGHVLAMIGKDWTHLGVAKLFSE ISRNGYNYLYLTARSAGQADSTRSYLRSIEQNGSKLPNGPVILSPDRTMAALRREVILKKPEVFKIACLNDIRSLYFEDSDNEMDTEEKSTPFFAGFGNRITDALSYRTVGIPSSRIFTINTEGEVHMELLELAGYRSSYIHINELVDHFFPPVSLDSVDLRTNTSMVPGSPPNRTLDNFDSEITSGRKTLFRGNQEEKFTDVNFWRDPLVDIDNLSDISNDDSDNIDEDTDVSQQSNVSRNRANSVKTAKVTKAPQRNVSGSTNNNEVLAASSDVENASDLVGSHSSSGSTPNKSTMSKGDIGKQIYLEGSPLASPKLRYLDDMDDEDSNYNRTKSRRASSAAATSIDKEFKKLSVSKAGAPTRIVSKIDVSNDVHSLGNSDTESRREQSVNETGRNQLPHNSMDDKDLDSRVSDEFDDDEFDEDEFED SEQ ID NO.96, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PCD1, amino acid sequence ColumnMILSQRRMLSSKQLIENLIRYKFHKTPYTRSSIWPFKRNSAVIILLFIGMKGELRVLLTKRSRTLRSFSGDVSFPGGKADYFQETFESVARREAEEEIGLPHDPEVLHKEFGMKLDNLVMDMPCYLSRTFLSVKPMVCFLYKDKLEKHEDKYKVPLDIRKFFGKLNPGETSSLFSVPLNDLVIHLLPEADEDVKSYQAEYFERKEYKLNWGGIKWLIMHYHFHVANNNEMPWLQTIEDLSSSDEDGVDGGIFRFRDLWGLTCKILFDVSCIANGLMDEKLKGELGHEDLIVGLHDYGNQMQPNGRSEWEIGMINGDRNLKYSDVIPEYYMKHLLECRSLW SEQ ID NO.97, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PDL32, amino acid sequence Column MTGSLNRHSLLNGVKKMRIILCDTNEVVTNLWQKSIPHAYIQNDKYLCIHHGHLQSLMDSMRKGDAIHHGHSYAIVSPGNSYGYLGGGFDKALYNYFGGKPFETWFRNQLGGRYHTVGSATVVDLQRCLEEKTIECRDGIRYIIHVPTVVAPSAPIFNPQNPLKTGFEPVFNAMWNALMHSPKDIDGLIIPGLCTGYAGVPPIISCKSMAFALRLYMAGDHISKELKNVLIMYYLQYPFEPFFPESCKTECQKLGIDIEMLKSFNVEKDAIELLIPRRILTLNL SEQ ID NO.98, derived from brewer's yeast ( Saccharomyces cerevisiae PEP4, amino acid sequence ColumnMFSLKALLPLALLLVSANQVAAKVHKAKIYKHELSDEMKEVTFEQHLAHLGQKYLTQFEKANPEVVFSREHPFFTEGGHDVPLTNYLNAQYYTDITLGTPPQNFKVILDTGSSNLWVPSNECGSLACFLHSKYDHEASSSYKANGTEFAIQYGTGSLEGYISQDTLSIGDLTIPKQDFAEATSEPGLTFAFGKFDGILGLGYDTISVDKVVPPFYNAIQQDLLDEKRFAFYLGDTSKDTENGGEATFGGIDESKFKGDITWLPVRRKAYWEVKFEGIGLGDEYAELESHGAAIDTGTSLITLPSGLAEMINAEIGAKKGWTGQYTLDCNTRDNLPDLIFNFNGYNFTIGPYDYTLEVSGSCISAITPMDFPEPVGPLAIVGDAFLRKYYSIYDLGNNAVGLAKAI SEQ ID NO.99, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHM8, amino acid sequence Column MTIAKDYRTIYRNQIKKQIRLNQEHLQSLTHLGSQINFEVDPPKLPDPDPARKVFFFDIDNTLYRKSTKVQLLMQQSLSNFFKYELGFDDDEAERLIESYYQEYGLSVKGLIKNKQIDDVLQYNTFIDDSLPLQDYLKPDWKLRELLINLKKKKLGKFDKLWLFTNSYKNHAIRCVKILGIADLFDGITYCHYDRPIEEEFICKPDPKFFETAKLQSGLSSFANAWFIDDNESNVRSALSMGMGHVIHLIEDYQYESENIVTKDHKNKQQFSILKDILEIPLIMDVEVYRPSSIAIKEMEELEEEGEAVNWSNQQINVQSS SEQ ID NO.100, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PHO11, amino acids SequenceMLKSAVYSILAASLVNAGTIPLGKLSDIDKIGTQTEIFFPLGGSGPYYSFPGDYGISDRLPESCEMKQVQMVGRHGERYPTVSKASIMTTWYKLSNYTGQFSGALSFLNDDYEFFIRDTKNLEMETTLANSVNVLNPYTGEMNAKRHARDFLAQYGYMVENQTSFAVFTSNSRCHDTAQYFIDGLDKFNISLQTISEAESAGANTLSAHHSCPAWDDDVNDDILKKYDTKYLSGIAKRLNKENKGLNLTSSDANTFFAWCAYEINARGYSDICNIFTKDELVRFSYGQDLETYYQTGPGYDVVRSVGANLFNASVKLLKESEVQDQKVWLSFTHDTDILNYLTTIGIIDDKNNLTAEHVPFMENTFHRSWYVPQGARVYTEKFQCSNDTYVRYVINDAVVPIETCSTGPGFSCEINDFYDYAEKRVAGTDFLKVCNVSSVSNSTELTFFWDWNTKHYNDTLLKQ SEQ ID NO.101, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PHO12, amino acids Sequence MLKSAVYSILAASLVNAGTIPLGKLSDIDKIGTQTEIFFPLGGSGPYYSFPGDYGISDRLPESCEMKQVQMVGRHGERYPTVSKASIMTTWYKLSNYTGQFSGALSFLNDDYEFFIRDTKNLEMETTLANSVNVLNPYTGEMNAKRHARDFLAQYGYMVENQTSFAVFTSNSRCHDTAQYFIDGLDKFNISLQTISEAESAGANTLSAHHSCPAWDDDVNDDILKKYDTKYLSGIAKRLNKENKGLNLTSSDANTFFAWCAYEINARGYSDICNIFTKDELVRFSYGQDLETYYQTGPGYDVVRSVGANLFNASVKLLKESEVQDQKVWLSFTHDTDILNYLTTIGIIDDQNNLTAEHVPFMENTFHRSWYVPQGARVYTEKFQCSNDTYVRYVINDAVVPIETCSTGPGFSEINDFYGYAEKRVAGTDFLKVCNVSSVSNSTELTFFWDWNTKHYNDTLLKQ SEQ ID NO.102, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHO13, amino acidSequence MTAQQGVPIKITNKEIAQEFLDKYDTFLFDCDGVLWLGSQALPYTLEILNLLKQLGKQLIFVTNNSTKSRLAYTKKFASFGIDVKEEQIFTSGYASAVYIRDFLKLQPGKDKVWVFGESGIGEELKLMGYESLGGADSRLDTPFDAAKSPFLVNGLDKDVSCVIAGLDTKVNYHRLAVTLQYLQKDSVHFVGTNVDSTFPQKGYTFPGAGSMIESLAFSSNRRPSYCGKPNQNMLNSIISAFNLDRSKCCMVGDRLNTDMKFGVEGGLGGTLLVLSGIETEERALKISHDYPRPKFYIDKLGDIYTLTNNEL SEQ ID NO.103, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHO2, amino acid sequence Column MMEEFSYDHDFNTHFATDLDYLQHDQQQQQQQHDQQHNQQQQPQPQPIQTQNLEHDHDQHTNDMSASSNASDSGPQRPKRTRAKGEALDVLKRKFEINPTPSLVERKKISDLIGMPEKNVRIWFQNRRAKLRKKQHGSNKDTIPSSQSRDIANDYERGSTDNNLVTTTSTSSIFHDEDLTFFDRIPLNSNNNYYFFDICSITVGSWNRMKSGALQRRNFQSIKELRNLSPIKINNIMSNATDLMVLISKKNSEINYFFSAMANNTKILFRIFFPLSSVTNCSLTLETDDDIINSNNTSDKNNSNTNNDDDNDDNSNEDNDNSSEDKRNAKDNFGELKLTVTRSPTFAVYFLNNAPDEDPNLNNQWSICDDFSEGRQVNDAFVGGSNIPHTLKGLQKSLRFMNSLILDYKSSNEILPTINTAIPTAAVPQQNIAPPFLNTNSSATDSNPNTNLEDSLFFDHDLLSSSITNTNNGQGSNNGRQASKDDTLNLLDTTVNSNNNHNANNEENHLAQEHLSSDADIVANPNDHLLSLPTDSELPNTPDFLKNTNELTDEHRWI SEQ ID NO.104, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHO3, amino acid sequence ColumnMFKSVVYSVLAAALVNAGTIPLGELADVAKIGTQEDIFPFLGGAGPYFSFPGDYGISRDLPEGCEMKQLQMLARHGERYPTYSKGATIMKTWYKLSNYTRQFNGSLSFLNDDYEFFIRDDDLEMETTFANSDNVLNPYTGEMDAKRHAREFLAQYGYMFENQTSFPIFAASSERVHDTAQYFIDGLDQFNISLQTVSEAMSAGANTLSAGNACPGWDEDANDDILDKYDTTYLDDIAKRLNKENKGLNLTSKDANTLFAWCAYELNARGYSDVCDIFTEDELVRYSYGQDLVSFYQDGPGYDMIRSVGANLFNATLKLLKQSETQDLKVWLSFTHDTDILNYLTTAGIIDDKNNLTAEYVPFMGNTFHKSWYVPQGARVYTEKFQCSNDTYVRYVINDAVVPIETCSTGPGFSEINDFYDYAEKRVAGTDFLKVCNVSSVSNVTELTFYWDWNTTHYNDTLLKQ SEQ ID NO.105, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHO5, amino acid sequence Column MFKSVVYSILAASLANAGTIPLGKLADVDKIGTQKDIFPFLGGAGPYYSFPGDYGISRDLPEGCEMKQLQMVGRHGERYPTVSLAKTIKSTWYKLSNYTRQFNGSLSFLNDDYEFFIRDDDLEMETTFANSDDVLNPYTGEMNAKRHARDFLAQYGYMVENQTSFAVFTSNSKRCHDTAQYFIDGLDQFNITLQTVSEAESAGANTLSACNSCPAWDYDANDDIVNEYDTTYLDDIAKRLNKENKGLNLTSTDASTLFSWCAFEVNAKGYSDVCDIFTKDELVHYSYYQDLHTYYHEGPGYDIIKSVGSNLFNASVKLLKQSEIQDQKVWLSFTHDTDILNFLTTAGIIDDKNNLTAEYVPFMGNTFHRSWYVPQGARVYTEKFQCSNDTYVRYVINDAVVPIETCSTGPGFSEINDFYDYAEKRVAGTDFLKVCNVSSVSNSTELTFYWDWNTTHYNASLLRQ SEQ ID NO.106, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PHO8, amino acid sequenceColumn MMTHTLPSEQTRLVPGSDSSSRKPKRRISKRSKIIVSTVVCIGLLLVLVQLAFPSSFALRSASHKKKNVIFFVTDGMGPASLSMARSFNQHVNDLPIDDILTLDEHFIGSSRTRSSDSLVTDSAAGATAFACALKSYNGAI GVDPHHRPCGTVLEAAKLAGYLTGLVVTTRITDATPASFSSHVDYRWQEDLIATHQLGEYPLGRVVDLLMGGGRSHYPQGEKASPYGHHGARKDGRDLIDEAQSNGWQYVGDRKNFDSLLKSHGENVTLPFLGLFADNDIP FEIDRDEKEYPSLKEQVKVALGALEKASNEDKDSNGFFLMVEGSRIDHAGHQNDPASQVREVLAFDEAFQYVLEFAENSDTETVLVSTSDHETGGLVTSRQVTASYPQYVWYPQVLANATHSGEFLKRKLVDFVHEHKGASSKIENFIKHEILEKDLGIYDYTDSDLETLIHLDDNANAIQDKLNDMVSFRAQIGWTTHGHSAVDVNIYAYANKKATWSYVLNNLQGNHENTEVGQFLENFLELNLNEVTDLIRDTKHTSDFDATEIASEVQHYDEYYHELTN SEQ ID NO.107, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PHO8 ^62aa, ammonia Amino acid sequenceMSHKKKNVIFFVTDGMGPASLSMARSFNQHVNDLPIDDILTLDEHFIGSSRTRSSDSLVTDSAAGATAFACALKSYNGAIGVDPHRPCGTVLEAAKLAGYLTGLVVTTRITDATPASFSSHVDYRWQEDLIATHQLGEYPLGRVVDLLMGGGRSHFYPQGEKASPYGHHGARKDGRDLIDEAQSNGWQYVGDRKNFDSLLKSHGENVTLPFLGLFADNDIPFEIDRDEKEYPSLKEQVKVALGALEKASNEDKDSNGFFLMVEGSRIDHAGHQNDPASQVREVLAFDEAFQYVLEFAENSDTETVLVSTSDHETGGLVTSRQVTASYPQYVWYPQVLANATHSGEFLKRKLVDFVHEHKGASSKIENFIKHEILEKDLGIYDYTDSDLETLIHLDNANAIQDKLNDMVSFRAQIGWTTHGHSAVDVNIYAYANKKATWSYVLNNLQGNHENTEVGQFLENFLELNLNEVTDLIRDTKHTSDFDATEIASEVQHYDEYYHELTN SEQ ID NO.108, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PIG1, amino acid sequence ColumnMPYSHGKKLKPSLKLAKTISTSSFVSSTTSNSFSPLEDSTSASLSTSSSSSGKSVRFAAHLYTVKKFNTKLAPISISEKAASNLTRNLHNNAIPLTFPFIGGEDHRYSLDILDYSDLEYDNKDVEYDNESDVEDNAMLMHDRSMFIEKEILCFGEEETFDMADWKLVSNNLNPFKSDCKVDVTELEDKIFKYLNGQNIKVHSLELSDPVSYEDICSNNFGNCQIWGLIFVNNLNFEKKIEIKFTLNNWADIHYINAHYNKSVTPHVDEFKFIIDISALKLNLISKNLIYANFFERKTTCLLNLQFCCRYDVNGFDYRSFYDNNDYKNYEITISLSAINLNRAVSNSSIFNSNLGPSKMRASNAEVTMSKNNKNSKKPLRKFIKDTDYYNDSPLKHKFYQSFETKAACKTEPVPQTFKAETIDCEIEPFNYFFEPPDSQTNEDMSDSSYDLSLQDFNYWEFSNHGLGKALADSDILQFKNYPKPEPFSRPPIIDDTFTLNTDDRTLGLKTEKLEDNLAKEWKSAKTRTTLNETPLHDDEHRTSFTYTTWNNSTDTLMKKQEERPVESASCSQLSIATIKAEEDLLYQDYINSGRESSSPEISPLNNTTSLPFFPGDNMSDSSGEYKERISLSPNKIHIFRDYFYKSPSP SEQ ID NO.109, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PIG2, amino acid sequence ColumnMATTTQPQNILMDEPLNLPNNSAHNNNYGNINANIRTSAGMSMHMHPARLNSLEFLHKPRRLSNVKLHRLPQDELQRNTDMNKGMYFNGKQVHAHHPFINSGANFNAHHQDVSKLGEEEDEISPLSHDNFQYESEENGNPSPPIYKKSGELVKSSLKRRSKSLPITPKSIFNKTGSKSKHVNLDHVDTRLLQRSKSVHFDRVLPIKLFNENEKPIDVGKQMVQQDVLNFKHKPLTRLSALNGGSDSVPIEDLLSENNQNEYGDTWLQNPKGVFLFGTNSNNRRNKKKKFKLSDDDSDIENDNDSDDAINRLVRQQDKDQAHLAHGLKNLLINDDDDYLETRTNSAKSGANLFIGNSKRIVGLYNKNFPILSDRDRKSLKLNIFLNLSRGRPVFLQEITLLTGFHNMVIIGKVFVKNIYFDKKIIVRYTWDAWRTFHESECVYFSNANGILPGSNMDIFKFSIDDIHNPNDKDSNISQLEFCIQYLTWGVDRSRKEYWDNNDSANYKIDVVTNETRTGPTTDVNDNYEMKHSLFRNPFH SEQ ID NO. 110, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PIK1, amino acid sequence Column SEQ ID NO.111, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PMU1, amino acid sequence Column MSLRAVPGYFAAYPSEGFQGLDSTKYDHLELINHKNWKELYHAIPRNTKNRHYKLLILARHGQGYHNAAILRYGMEKWDAYWSLLSGDEHGEWLDSKLTPLGKDQVRRTGSNVLLPMAKQLGMLPHVFFSSPMRRCLETFIESWTPVLAETQELPAGTKISTRIIEGLRETLGSHTCDKRVAHSMAVDEYQDFSTESGHTVHWQYVPDYPEDDELWLPDHRETCAEMDKRTLNGLFELFNQLSSEEKFISLTCHSGVIQSVLRNLQHPPIYNLDTGKVVAVVVEVPVNTADRGRL SEQ ID NO.112, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae POA1, amino acid sequence Column MSNITYVKGNILKPKSYARILIHSCNCNGSWGGGIAYQLALRYPKAEKDYVEVCEKYGSNLLGKCMLLPSYENSDLLICCLFTSSFGGSSHGEKQSILNYTKLALDKLKTFREAKDKTRTSEDIIGDYLNGHIKYPIGEYKLEMPQINSGIFGVPWKETERVLEEFSGDMSFTVYQL SEQ ID NO.113, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPA2, amino acid sequence Column MNLLRMNALTSKTRSIERLKQTLNILSIRNHRQFSTIQQGSKYTLGFKKYLTLLNGEVGSFFHDVPLDLNEHEKTVNMIVEVPRWTTGKFEISKELRFNPIVQDTKNGKLRFVNNIFPYHGYIHNYGAIPQTWEDPTIEHKLGKCDVALKGDNDPLDCCEIGSDVLEMGSIKKVKVLGSLALIDNGELDWKVIVIDVNDPLSSKIDDLEKIEEYFPGILDATREWFRKYKVPAGKPLNSFAFHEQYQNSNKTIQTIKECHNSWKKLISGSLQEKYDNLPNTERAGNGVTLEDSVKPPSQIPPEVQKWYYV SEQ ID NO.114, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPE1, amino acid sequence ColumnMSDDLRRKIALSQFERAKNVLDATFQEAYEDDENDGDALGSLPSFNGQSNRNRKYTGKTGSTTDRISSKEKSSLPTWSDFFDNKELVSLPDRDLDVNTYYTLPTSLLSNTTSIPIFIFHHGAGSSGLSANLAKELNTKLEGRCGCFAFDARGHAETKFKKADAPICFDRDSFIKDFVSLLNYWFKSKISQEPLQKVSVILIGHSLGGSICTFAYPKLSTELQKKILGITMLDIVEEAAIMALNKVEHFLQNTPNVFESINDAVDWHVQHALSRLRSSAEIAIPALFAPLKSGKVVRITNLKTFSPFWDTWFTDLSHSFVGLPVSKLLILAGNENLDKELIVGQMQGKYQLVVFQDSGHFIQEDSPIKTAITLIDFWKRNDSRNVVIKTNGQHKTVQNT SEQ ID NO.115, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPG1, amino acid sequence Column MELDECLERLYKAQLLPEVTVRALCFKLKEMLVKESNVIHIQTPVTVVGDMHGQFHDMLEIFQIGGPVPDTNYLFLGDYVDRGLYSVETIMLLIVLKLYPSRIHLRGNHESRQITQSYGFYTECLNKYGGNSRVWQYLTDIFDYLLVLCCIIDDEIFCVHGGLSPNVQTIDQIKIIDRFREIP HDGAMADLVWSDPEENNNPTLDHPDNGSQHFQVSPRGAGYTFGRSVVEKFLRMNDMNRIIRAHQLCNEGYQIYFDGLVTTVWSAPNYCYRCGNKASILELYSKDQFYFNVFEEAPENKLLKENSMNDNALEDSISNPVANRKLIADYFEDDSASADGSTDPEMYIFSDVYQARSASNRHVDYFL SEQ ID NO.116, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPH21, amino acids SequenceMDTDLDVPMQDAVTEQLTPTVSEDMDLNNNSSDNNAEEFSVDDLKPGSSGIADHKSSKPLELNNTNINQLDQWIEHLSKCEPLSEDDVARLCKMAVDVLQFEENVKPINVPVTICGDVHGQFHDLLELFKIGGPCPDTNYLFMGDYVDRGYYSVETVSYLVAMKVRYPHRITILRGNHESRQITQVYGFYDECLRKYGSANVWKMFTDLFDYFPITALVDNKIFCLHGGLSPMIETIDQVRELNRIQEVPHEGPMCDLLWSDPDDRGGWGISPRGAGFTFGQDVSEQFNHTNDLSLIARAHQLVMEGYAWSHQQNVVTIFSAPNYCYRCGNQAAIMEVDENHNRQFLQYDPSVRPGEPSVSRKTPDYFL SEQ ID NO.117, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPH22, amino acids Sequence MAVDVLQFEENVKPINVPVTICGDVHGQFHDLLELFKIGGPCPDTNYLFMGDYVDRGYYSVETVSYLVAMKVRYPHRITILRGNHESRQITQVYGFYDECLRKYGSANVWKMFTDLFDYFPITALVDNKIFCLHGGLSPMIETIDQVRELNRIQEVPHEGPMCDLLWSDPDDRGGWGISPRGAGFTFGQDVSEQFNHTNDLSLIARAHQLVMEGYSWSHQQNVVTIFSAPNYCYRCGNQAAIMEVDENHNRQFLQYDPSVRPGEPTVTRKTPDYFL SEQ ID NO.118, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPH3, amino acid sequence ColumnMMDLDKIIASLRDGKHIPEETVFRCLNSQELLMNEGNVTQVDTPVTICGDIHGQLHDLLTLFEKSGGVEKTRYIFLGDFVDRGFYSLESFLLLLCYKLRYPDRITLIRGNHETRQITKVYGFYDEVVRKYGNSNVWRYCCEVFDYLSLGAIINNSIFCVHGGLSPDMTTVDEIRTIDRKQEVPHEGAMCDLLWSDPEDDVDTWSLSPRGAGFLFGKREVDQFLEKNNVELIARAHQLVMEGYKEMFDGGLVTWSAPNYCYRCGNVAAVLKIDDDLNREYTIFEAVQAQNEVGNAIIPTKKSQMDYFL SEQ ID NO.119, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPM1, amino acid sequence Column MERIIQQTDYDALSCKLAAISVGYLPSSGLQRLSVDLSKKYTEWHRSYLITLKKFSRRAFGKVDKAMRSSFPVMNYGTYLRTVGIDAAILEFLVANEKVQVVNLGCGSDLRMLPLQMFPHLAYVDIDYNESVELKNSILRESEILRISLGLSKEDTAKSPFLIDQGRYKLAACDLNDITETTRLLDVCTKREIPTIVISECLLCYMHNNESQLLINTIMSKFSHGLWISYDPIGGSQPNDRFGAIMQSNLKESRNLEMPTLMTYNSKEKYASRWSAAPNVIVNDMWEIFNAQIPESERKRLRSLQFLDELEELKVMQTHYILMKAQW SEQ ID NO.120, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPM2, amino acid sequence ColumnMKNLTTIKQTNKNVKQERRKKYADLAIQGTNNSSIASKRSVELLYLPKLSSANNFQMDKNNKLLEYFKFFVPKKIKRSPCINRGYWLRLFAIRSRLNSIIEQTPQDKKIVVVNLGCGYDPLPFQLLDTNNIQSQQYHDRVSFIDIDYSDLLKIKIELIKTIPELSKIIGLSEDKDYVDDSNVDFLTTPKYLARPCDLNDSKMFSTLLNECQLYDPNVVKVFVAEVSLAYMKPERSDSIIEATSKMENSHFIILEQLIPKGPFEPFSKQMLAHFKRNDSPLQSVLKYNTIESQVQRFNKLGFAYVNVGDMFQLWESADEATKKELLKVEPFDELEEFHLFCHHYVLCHATNYKEFAFTQGFLFDRSISEINLTVDEDYQLLECECPINRKFGDVDVAGNDVFYMGGSNPYRVNEILQLSIHYDKIDMKNIEVSSSEVPVARMCHTFTTISRNNQLLLIGGRKAPHQGLSDNWIFDMKTREWSMIKSLSHTRFRHSACSLPDGNVLILGGVTEGPAMLLYNVTEEIFKDVTPKDEFFQNSLVSAGLEFDPVSKQGIILGGGFMDQTTVSDKAIIFKYDAENATEPITVIKKLQHPLFQRYGSQIKYITPRKLLIVGGTSPSGLFDRTNSIISLDPLSETLTSIPISRRIWEDHSLMLAGFSLVSTSMGTIHIIGGGATCYGFGSVTNVGLKLIAIAK SEQ ID NO.121, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPN2, amino acid sequence ColumnMEDKRKRRAATLSTALILFVACVYTLYIFKFDNPRLSPPVSLLPTISTLKIEHVTDLNKEYVFVGDVHGNYDEFIELIDDKIGGGLGENITMILLGDFIHKGPDSDKVVSYILNHKDQVKCVLGNHEILVMMAYLNPDFSKWVRRPKLMTPLTFSTETNFIPQDISKISNAHGRLARELGFSKLSQLAEHCSMAIELDLDITGDILFGAHAGMVPDGDFMKPNQIPGVSSLSNMKYVDKKNWSKTSREKENKNYVRWYTLWDKYGDHFSNAKVFYGHDASMGLNLRRQTKGLDTACIKNNLLSSMKVKYDIKKGQYDYELIQVQCS SEQ ID NO.122, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPQ1, amino acid sequence Column MRRSPSRSNNNFAVPNCSTNSNSSQQQLTTPSDDLNSNEPNDPDDSRSLPTIKKFNNKHSINNYNTLASAGKNNNNKRASNDNLLIPGENAHKQKIYTKDENLKSLYLDIDVSVAKALSSSATAPKLINTARTSSTTTATTSNNILTSPSSYRESNYSSPSSYSFSSYYSSATSASSSTSSFLKSSGLSSRVKSPSSSVKAGSFGAPSSPTSGIPNPKSSKKPIFLRRYSHDTSSNEGLDIDVAIEKLLQVGESREITKTSKKKNFPFHSWEIQL ICYHAREIFLNQPTLLRLQAPIKVVGDVHGQFNDLRILKLSGVPSDTNYLLFLGDYVDRGKNSLETILLLLCYKIKYKDNFFMLRGNHESANVTKMYGFYDECKRRLSSKVWKMFVDVFNTLPLAAIIQDKIFCVHGGISPDLHDMKQIEKVARPTDIPESGLVTDLLWSDPDPQVTDWSENDRGVSYTFSKRNVLDFCAKFKFDLILRGHMVVEDGYEFFARKKFVTIFSAPNYCGEFHNWGAVMSVTTGMMCSFELLKPRALKNKKKLYKTKV SEQ ID NO.123, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPS1, amino acid sequence ColumnMVLEVPSITPGELHDLMRLHQDAEWPECKKMFPWAHDISFGQPPDFPHSLAIVKSQSDANNSALLRNSLEVNDIFQSWKVRTSFHREGDTCETGNDSNGFQYPNNTKELLNLLKFQIRQLELQVDDVALENAATYCHNHSILPFLKVDPRGLSLELKRYSRNKVGSNTTLKRSGQDVWGRRGLFRRFDLQCAKMIEMVDNIVIYCSRTGGSTDMQTESAPACSHEGNCPNCTTLALLLQICLMFVQKGYVGSGGSLYKTNLFICTYQNFNTDIPQTLIGTPLLDNEFFKNNTPLNLCSSPSEIVCFNNVDKNMVLCEKLELNKLTSATRLEETGLICGNTTDWHNYQIIKKNNISLTHRFEENTSIVNLKSLNYDTDNPTTSISQLYNIPNTKEVWKLIIKCTSNSQMPSLTKIRTYLDLLLDDDASKSQEHLHLTFPASGSIGLGNLNIQSVEILLNVCYLIFQVSQVQELLTFMYCEDGYTETSLLLTAYIIFHFNIPLQDALLRIHPRPFFLFPSDLQILGHLQPVLREFSPQNGSNLKLYANALKFRDKSFQLHISSELFSSIFFMKIPLESNFVNLKGPLPSRILRHLYLGSLDHAQNPALLKSLGITHIVSVGEVVSWTLNKDKIAHPVRPHRAITMTNTNEVAGNTTCNKSRNRADTVVSDKQENGSNVVISENSGFQICQIENLDDNGKDPLFHQIDKVLDFISNSEATGGKVLVHCMVGVSRSATVCIAECMRYLQCDLASAYLFVRVRRLNVIIQPNLFFVYELFKWWKKHYNREKDKTMDWHIICRGIAEVNMKYT SEQ ID NO.124, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPT1, amino acid sequence ColumnMSTPTAADRAKALERKNEGNVFVKEKHFLKAIEKYTEAIDLDSTQSIYFSNRAFAHFKVDNFQSALNDCDEAIKLDPKNIKAYHRRALSCMALLEFKKARKDLNVLLKAKPNDPAATKALLTCDRFIREERFRKAIGGAENEAKISLCQTLNLSSFDANADLANYEGPKLEFEQLYDDKNAFKGAKIKNMSQEFISKMVNDLFLKGKYLPKKYVAAIISHADTLFRQEPSMVELENNSTPDVKISVCGDTHGQFYDVLNLFRKFGKVGPKHTYLFNGDFVDRGSWSCEVALLFYCLKILHPNNFFLNRGNHESDNMNKIYGFEDECKYKYSQRIFNMAQSFESLPLATLINNDYLVMHGGLPSDSPASTLSDFKNIDRFAQPPRDGAFMELLWADPQEANGMGPSQRGLGHAFGPDITDRFLRNNKLRKIRSKIRFSHELRMGGVQFEQKGKLMTVFSAPNYCDSQGNLGGVIHVVPGHGILQAGRNDDQNLIIETFEAVEHPDIKPMAYSNGGFGL SEQ ID NO.125, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PPZ1, amino acid sequence ColumnMGNSSSKSSKKDSHSNSSSRNPRPQVSRTETSHSVKSAKSNKSSRSRRSLPSSSTTNTNSNVPDPSTPSKPNLEVNHQRHSSHTNRYHFPSSSHSHSNSQNELLTTPSSSSTKRPSTSRRSSYNTKAAADLPPSMIQMEPKSPILKTNNSSTHVSKHKSSYSSTYYENALTDDDNDDKDNDISHTKRFSRSSNSRPSSIRSGSVSRRKSDVTHEEPNNGSYSSNNQENYLVQALTRSNSHASSLHSRKSSFGSDGNTAYSTPLNSPGLSKLTDHSGEYFTSNSTSSLNHHSSRDIYPSKHISNDDDIENSSQLSNIHASMENVNDKNNNITDSKKDPNEEFNDIMQSSGNKNAPKKFKKPIDIDETIQKLLDAGYAAKRTKNVCLKNNEILQICIKAREIFLSQPSLLELSPPVKIVGDVHGQYGDLLRLFTKCGFPPSSNYLFLGDYVDRGKQSLETILLLFCYKIKYPENFFLLRGNHECANVTRVYGFYDECKRRCNIKIWKTFIDTFNTLPLAAIVAGKIFCVHGGLSPVLNSMDEIRHVVRPTDVPDFGLINDLLWSDPTDSPNEWEDNERGVSYCYNKVAINKFLNKFGFDLVCRAHMVVEDGYEFFNDRSLVTVFSAPNYCGEFDNWGAVMSVSEGLLCSFELLDPLDSAALKQVMKKGRQERKLANQQQQMMETSITNDNESQQ SEQ ID NO.126, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PPZ2, amino acid sequence ColumnMGNSGSKQHTKHNSKKDDHDGDRKKTLDLPPLTKSDTTHSLKSSRSLRSLRSKRSEASLASNVQAQTQPLSRRSSTLGNGNRNHRRSNNAPITPPNNHYLTSHPSSSRRLSSSSRRSSMGNNNNSELPPSMIQMEPKSPILKNSTSMHSTSSFNSYENALTDDDDDRGDDGGESPSMAKVTRINTSSSADRGSKRTPLRRHNSLQPEKGVTGFSSTSSKLRRRSDNTLPASYPLNAEAGGNGSDYFSNRSNSHASSRKSSFGSTGNTAYSTPLHSPALRKMSSRDNDDSGDNVNGRGTSPIPNLNIDKPSPSASSASKREYLSAYPTLAHRDSSSSLSPRGKGQRSSSSSSSSQRIYVSPPSPTGDFVHGSCADGDNGSRTNTMVEMKRKKPVRPVDIDEIIQRLLDAGYAAKRTKNVCLKNSEIIQICHKARELFLAQPALLELSPSVKIVGDVHGQYADLLRLFTKCGFPPMANYLFLGDYVDRGKQSLETILLLLCYKIKYPENFFLLRGNHECANVTRVYGFYDECKRRCNIKIWKTFVDTFNTLPLAAIVTGKIFCVHGGLSPVLNSMDEIRHVSRPTDVPDFGLINDLLWSDPTDSSNEWEDNERGVSFCYNKVAINKFLNKFGFDLVCRAHMVVEDGYEFFNDRSLVTVFSAPNYCGEFDNWGAVMTVSEGLLCSFELLDPLDSTALKQVMKKGRQERKLANR SEQ ID NO.127, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PSR1, amino acid sequence ColumnMGFISSILCCSSETTQSNSNSAYRQQQSSSLNKNRSVKHSNTKSRTRGVHQTNSPPSKTNSAATFSSTERSTGKSGISTNDNEKKKQSSPTAAVTATTTNNMTKVEKRISKDDLYEEKYEVDEDEEIDDEDNRRSRGIVQEKGDAVKDTSRQKKQQQQQQQQSQSQPQPQPQSQSQSQSQSQSQSQSQQRGPTVQVSSDHLIQDMNLSRVSSSSQASETSNDVDDEDDEDEEYIDLTLLQQGQYHAPGYNTLLPPQDESTKGKKCLILDLDETLVHSSFKYLRSADFVLPVEIDDQVHNVYVIKRPGVEEFLERVGKLFEVVVFTASVSRYGDPLLDILDTDKVIHHRLFREACYNYEGNYIKNLSQIGRPLSDIIILDNSPASYIFHPQHAIPISSWFSDTHDNELLDIIPLLEDLSVKTSLDVGKILDVTI SEQ ID NO.128, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PSR2, amino acid sequence Column MGFIANILCCSSDTSKTHRQRQPPETNHNRNRNRKHSSNKAQTQGRKQKATPNGDKMQYSTPKILLSSSDSGSNAGSKTMQENGNSGNGKLAPLSRDHSNNSYDEEKEYEDYNEGDVEMTEVNNAGEEEEEDDEAKEKQDHVVHEYNVDADRNSSINDEAPPQQGLYQVDQEDMNPQYVASSPDNDLNLIPTTEEDFSDLTHLQPDQYHAPGYDTLLPPKLQEFKQKKCLILDLDETLVHSSFKYMHSADFVLPVEIDDQVHNVYVIKRPGVDEFLNRVSQLYEVVVFTASVSRYANPLLDTLDPNGTIHHRLFREACYNYEGNYIKNLSQIGRPLSETIILDNSPASYIFHPQHAVPISSWFSDTHDNELLDIIPLLEDLSSGNVLDVGSVLDVTI SEQ ID NO.129, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PSY2, amino acid sequence ColumnMSLPGTPTTSPTPMDEDTEQAVSVNTEPKRVKVYILENNEWKDTGTGFCIGEVDEGKFAYLVVSDEDSPTETLLKSKLEGNIEYQRQEETLIVWKDLGGKDIALSFEESMGCDTLCEFIVHVQRNIESNISLVTVKSSDNGLGSVHDIITGPVTLPSNDQQQNSQTLLEALKILNENTSFDFLKNETIEFILQSNYIDTLISHFHKAEEEKIPKDLFLLSNIIKTLILYNKRDILESMVEDDRIMGIVGILEYDTEYPTSKANHRKYLGSKGPNFKEVIPLENEDLKIIMKKCFRLQFLKDVVLVRFLDDHNFNLISEIVMDLETCIIDFLQVGTFLDRLIELYDTKTLPESSSEKFVQKRKDGIRLLQQCVQMSINLDAVDRSKFYKTLVRKGLFKVLDYAFHMETDSNVRILATDTIIIHEHDI LLIHNVQNEDSFKRQHKSAPDDKSSHRKYPQDYSSSTDSKLLILSTILLSDRSPGLREQVVQALNTLLHPEGCVGNGEGSYDLMGRSNYEAKNTSEDFPSFSYGLNSDSINLNNYHYSSDEMNNLEPESESEFQVMEYFANFYNKIAPILFGPLIKKDITTEMAEIDGQIEKVTKDDLLLIHLVKLVSFVCTEHDRVLSRRFILENGILDSVSKLIGGNHMMQLRLTAVRCIKNLMCLDDKYYHRYMISKNLYAPVFKLFQENIDKNNLANSCIQDFFRIIITECRAYQSDGHNRKEKTNGSYDGNGNDVKTNVNNNRTNFTILNKYLVQTYGDVLRKATDIPFIQDMLETGEENQPDHSSFENSIEGGNDISVNMSTDGFASNHLEDIDIKNVKRLHSEIEHFENDPHYSGDQLAFKKSVDQMNAST SEQ ID NO.130, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PSY4, amino acid sequence ColumnMGIKSISLYELLSDVVKQGDKTRLVTAGPEQVLPDLIRHITETIPFDLFINLKNEMNDARNLVTRLNWLGKFLNDFLQNHTFPFTILRICELCYDPFKYYKINELEKFVNALEKCCMVTSSWQVFDKTHGEKQEDDKEKDINFIKNQEDVSLMKIPWMTENNTRELAPFIREIDSIMSVNLGYDDEEDEDNGDGEEEGFFDGDEDREMGNSKRN VLLKDENFMVEEYYEDDCGINDDNTDNKGQNCQSDVTKNNSDDEDDDDNDDDYREDGADEDDDHMGSTDDDEDDEDRQAGESTKVQNFDKKNETPRKRKPTDLDNFEYDESPSFTNMDLTTPKKYKHTATGRFSIIESPSSSLLNAMDGSNEISSSQEEKEDARENHEGGSEGLLPGDELVSPTMSSSQEDKMVAIAGITYRENISSPLGKKSR SEQ ID NO.131, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PTC1, amino acid sequence Column MSNHSEILERPETPYDITYRVGVAENKNSKFRRTMEDVHTYVKNFASRLDWGYFAVFDGHAGIQASKWCGKHLHTIIEQNILADETRDVRDVLNDSFLAIDEEINTKLVGNSGCTAAVCVLRWELPDSVSDDSMDLAQHQRKLYTANVGDSRIVLFRNGSIRLTYDHKASDTLEMQRVEQAGGLIMKSRVNGMLAVTRSLGDKFFDSLVVGSPFTTSVEITSEDKFLILACDGLWDVIDDQDACELIKDITEPNEAKVLVRYALENGTTDNVTVMVVFL SEQ ID NO.132, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PTC2, amino acid sequence ColumnMGQILSNPVIDKESHGADSLTAFGLCAMQGWRMSMEDSHILEPNVLTKSDKDHIAFYGIFDGHGGAKVAEYCGNKIVEILQEQKSFHEGNLPRALIDTFINTDVKLLQDPVMKEDHSGCTATSILVSKSQNLLVCGNAGDSRTVLATDGNAKALSYDHKPTLASEKSRIVAADGFVEMDRVNGNNLALSRAIGDFEFKSNPKLGPEEQIVTCVPDILEHSLDYDRDEFVILACDGIWDCLTSQDCVDLVHLGLREGKTLNEISSRIIDVCCAPTTEGTGIGCDNMSIVVALLKEGEDVAQWSDRMKSKAHRTSVRSFADKRRRVFSYYDFSKCNDEQVFAITTKKPQDKFTRDHEAAVASVTAADNDDPMDIDDTDADTDAENLDPSSQSKSKTSGPIDLASLEALLGATGGVKTDSNGNKVTYTLPQSALAQLLQTMGHDPASSHPENDSNTDHKAGRSHLQ SEQ ID NO.133, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PTC3, amino acid sequence Column MGQILSNPIIDKEHHSGTDCLTAFGLCAMQGWRMSMEDAHIVEPNLLAESDEEHLAFYGIFDGHGGSSVAEFCGSKMISILKKQESFKSGMLEQCLIDTFLATDVELLKDEKLKDDHSGCTATVILVSQLKKLLICANSGDSRTVLSTGGNSKAMSFDHKPTLLSEKSRIVAADGFVEMDRVNGNLALSRAIGDFEFKSNTKLGPHEQVVTCVPDIICHNLNYDEDEFVILACD GIWDCLTSQECVDLVHYGISQGNMTLSDISSRIVDVCCSPTTEGSGIGCDNMSISIVALLKENESESQWFERMRSKNYNIQTSFVQRRKSIFDFHDFSDDDNEVFAITTKKLQDRLNRSKDNDDMEIDDLDTELGSSATPSKLSGEDRTGPIDLFSLEALLEAGIQIRQRPSSSDGNTSYFHGASLSDMLASLSNAAAGETPNDADDNDDNDGEENGKNENAKKGSKIEEIE SEQ ID NO.134, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PTC4, amino acid sequenceColumn MGQLLSHPLTEKTIEYNEYKNNQASTGIVPRFYNCVGSMQGYRLTQEDAHLIRNENSVVYVRFFNPFIDKYETLSLNVFAVFDGHGGDDCSKFLSGGRHHRDGNGSSNGEPNAGLIKWIAYSFENHYTSTTNNDSSKFKRSFNTLEGLVSQIFKDAFILQDEELYRHFANSSCGSTAVVACIINEESLYVANCGDSRCILSKSNGIKTMSFDHKPQHIGELIRINDNGGTVSLGRVGGVLALSRAFSDFQFKRGVTYPHRRTKLTNITQNLTYGTPPQEAQVTVEPDVLMHKIDYSKDEFLVLACDGIWDIYNNKQLIHFIKYHLVSGTKLDTIITKLLDHGIAQANSNTGVGFDNMTAIIVVLNRKGETLQDWFNKMKTRLERERGLV SEQ ID NO.135, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PTC5, amino acid sequence ColumnMSPLTRTVAIKKTVVLSKCQSGREYTQKFLQRAYSTSHANSTYYSRTKLFISSHSKALNIALLSGSLLLTYSYYSPKKILSLDTINGIKDYSTNTSGNINMPSPNPKGTETQKSQRSQNDQSVLILNDSKIEAKLHDREESHFVNRGTGIFRYDVAQLPSNHPIEDDHVEQIITIPIESEDGKSIEKDLYFFGIFDGHGGPFTSEKLSKDLVRYVAYQLGQVYDQNKTVFHSNPNQLIDSAISKGFLKLDNDLVIESFRKLFQDPNNTNIANTLPAISGSCALLS LYNSTNSILKVAVTGDSRALICGLDNEGNWTVKSLSTDQTGDNLDEVRRIRKEHPGEPNVIRNGRILGSLQPSRAFGDYRYKIKEVDGKPLSDLPEVAKLYFRREPRDFKTPPYVTAEPITSAKIGENTKFMVMGSDGLFELLTNEEIASLVIRWMDKNMNLAPVKAEPGKLPKVIDVSEDKEAQRPAFRYKDNNSSSPSGSNPEYLIEDKNVATHLIRNALSAGGRKEYVSALVSIPSPMSRRYRDDLTVTVAFFGDSGTPSIVSNATSIVMNPEATTKPKPRL SEQ ID NO.136, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PTC6, amino acid sequence ColumnMRLGNAYAYCKPSQNVGLKLDLRGLPGYVGHATSRINRLENQDNYSIKMMRSWPNAYGSALNCSVFDGHGEKGAQLSQLLADKLCSSLDFPEPSWDKQDLKKLVQEYARRFPEGNYWKHKLSTFEKFYNKFIKNCNSKQELLLMKEGDSAILGQNGGRMIFDKMGNIIDKIALTELDLLRLFYGFARFDLDQCCGLGTAAGSTASSIFLYPYDDPNAPI DEGKDDDSWIISHGSGLLKLIVTQVGDSKIILCDQDGIAHALTTTTHHINSSRERHRLSIDPSRLDPAFGETRFLNNFANTRSFGDVAGKPYGISSEPDIFSFLVGNTLHLPRSERSKLPFNGDECFLALVTDGITNKLADQEVVDLITSTVNSWGLKKATPQFVAEETIKFIQAIATKHSDNATCVVVRLSNWGNWPNVDRTGPQRETKLMNAQSNETKLN SEQ ID NO.137, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PTC7, amino acid sequence Column MFANVGFRTLRVSRGPLYGMFIVLFIGVLIAKFAGQMLIDSETNFSHIIGSCSQIISFSKRTFYSSAKSGYQSNNSHGDAYSSGSQSGPFTYKTAVAFQPKDRDDLIYQKLKDSIRSPTGEDNYFVTSNNVHDIFAGVADGVGGWAEHGYDSSAISRELCKKMDEISTALAENSSKETLLTPKKIIGAAYAKIRDEKVVKVGGTTAIVAHFPSNGKLEVANLGDSWCGVFRDSKLVFQTKFQTVGFNAPYQLSIIPEEMLKEAERRGSKYILNTPRDADEYSFQLKKKKDIIILATDGVTDNIATDDIELFLKDNAARTNDELQLLSQKFVDNVVSLSKDPNYPSVFAQEISKLTGKNYSGGKEDDITVVVVRVD SEQ ID NO.138, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PTP1, amino acid sequence ColumnMAAAPWYIRQRDTDLLGKFKFIQNQEDGRLREATNGTVNSRWSLGVSIEPRNDARNRYVNIMPYERNRVHLKTLSGNDYINASYVKVNVPGQSIEPGYYIATQGPTRKTWDQFWQMCYHNCPLDNIVIVMVTPLVEYNREKCYQYWPRGGVDDTVRIASKWESPGGANDMTQFPSDLKIEFVNVHKVKDYYTVTDIKLTPTDPLVGPVKTVHHFYFDLWKDMNKPEEVVPIMELCAHSHSLNSRGNPIIVHCSAGVGRTGTFIALDHLMHDTLDFKNITERSRHSDRATEEYTRDLIEQIVLQLRSQRMKMVQTKDQFLFIYHAAKYLNSLSVNQ SEQ ID NO.139, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PTP2, amino acid sequence ColumnMDRIAQQYGSGKRDNNGNRMASSAISEKGHIQVNQTRTPGQMPVYRGETINLSNLPQNQIKPCKDLDDVNIRRNNSNRHSKILLLDLCAGPNTNSFLGNTNAKDITVLSLPLPSTLVKRSNYPFENLLKNYLGSDEKYIEFTKIIKDYDIFIFSDSFSRISSCLKTTFCLIEKFKKFICHFFPSPYLKFFLLEGSLNDSKAPSLGKNKKNCILPKLDLNLNVNLTSRSTLNLRINIPPPNDSNKIFLQSLKKDLIHYSPNSLQKFFQFNMPADLAPNDTILPNWLKFCSVKENEKVILKKLFNNFETLENFEMQRLEKCLKFKKKPLHQKQLSQKQRGPQSTDDSKLYSLTSLQRQYKSSLKSNIQKNQKLKLIIPKNNTSSSPSPLSSDDTIMSPINDYELTEGIQSFTKNRYSNILPYEHSRVKLPHSPKPPAVSEASTTETKTDKSYPMCPVDAKNHSCKPNDYINANYLKLTQINPDFKYIATQAPLPSTMDDFWKVITLNKVKVIISLNSDDELNLRKWDIYWNNLSYSNHTIKLQNTWENICNINGCVLRVFQVKKTAPQNDNISQDCDLPHNGDLTSITMAVSEPFIVYQLQYKNWLDSCGVDMNDIIKLHKVKNSLLFNPQSFITSLEKDVCKPDLIDDNNSELHLDTANSSPLLVHCSAGCGRTGVFVTLDFLLSILSPTTNHSNKIDVWNMTQDLIFIIVNELRKQRISMVQNLTQYIACYEALLNYFALQKQIKNALPC SEQ ID NO.140, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae PTP3, amino acid sequence ColumnMKDSVDCPSILPTDRTSVLSETSTLVGSSSHVYSKHAPMNSYHNSMNSNIYHSPKASSPLVSYKTSSPVLLKRATAPVLPSFKPKEQRYNKPQGCSLITAVELGKIIETLPDEKVLLLDVRPFTEHAKSIITNSIHVCLPSTLLRRKNFTFSKLLDNLTPSEQSVLKSKLAIDNLRIIIYDSTANQTESSVSLPCYGIASKLIEFDTNVKKTVSILMCGFPQFKILFPDHINTNTFNSDCISSAEPKSPKTNLMNSLHNTAPHMTATTPLSSPQMNLKLKVPDDSRSDHSNFSSSPSPRNVLSDSPMSSSSPISALFKFQLPAPQTNINQMFKFSQNEEIMDLETYLSAVNIKEEHERWYNNDSAKKSLQNFQFPKNQNSLEKDTNKDKLGFQIRYENLSKNYEKEVIDSVIPEWFQHLMSIPKIELVSQFQKLDFLEKRRLNHSVSFRKKENSFILEKPSSYPEQLTSTSSSTIMPPKFPDVNKVQKRSHSQPIFTQYSKYKSMLSLESDSDSESDNVIISSGVELGAKNRYKDIFPYEHSRVILKKGLQSSKGIKHSHSTSDGGILDNYINANYLSLPRFSVEQNSSFQTTSTTTRRVRYIATQAPMPSTVHDFYTCILNNGVPLVLSLTNDFENGIEKCYRYWQEGNYNGIHVKLLEKKILKMPSTTSMRKNTTGTQNSSLYSAGVHGNSSNYSTDNDNDNDNNNNNNNNSNIAVTAAACDDDDDDDDDDAILIRKILLTYHDQEKPYELLQIQVKNWPDLGTLLNPTSILQAINVKNHIIDTLFARNYYQNDQLPTILVHCSAGCGRTGTLCTIDSILSNFEMFEMLQKEFVKLKYPAKLFDPISWTINIFRKQRISMVQNINQFIFIYDCLLFYFRLRLDDITERTDGDGSNKDNISLSALIEQIEKLEILQTFVDDKLKELPQ SEQ ID NO.141, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae PYP1, amino acid sequence ColumnMVKAVIFTDFDGTVTLEDSNDYLTDTLGFGKEKRLKVFEGVLDDTKSFRQGFMEMLESIHTPFPECIKILEKKIRLDPGFKDTFEWAQENDVPVIVVSSGMKPIIKVLLTRLVGQESIHKIDIVSNEVEIDAHDQWKIIYKDESPFGHDKSRSIDAYKKKFESTLKAGEQRPVYFCGDGVSDLSAAKECDLLFAKRGKDLVTYCKKQNVPHEFDTFKDILASMKQVLAGEKTVAELMEN SEQ ID NO.142, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae QRI7, amino acid sequence Column MISIKGTGRFLLDNYRIWQRRASNRPIQLRKGYKVLAIETSCDDTCVSVLDRFSKSAAPNVLANLKDTLDSIDEGGIIPTKAHIHHQARIGPLTERALISNAREGIDLICVTRGPGMPGSLSGGLDFAKGLAVAWNKPLIGVHMLGHLLIPRMGTNGKVPQFPFVSLLVSGGHTTFVLSRAIDDHEILCDTIDIAVGDSLD KCGRELGFKGTMIAREMEKFINQDINDQDFALKLEMPSPLKNSASKRNMLSFSFSAFITALRTNLTKLGKTEIQELPEREIRSIAYQVQESVFDHIINKLKHVLKSQPEKFKNVREFVCSGGVSSNQRLRTKLETELGTLNSTSFFNFYYPPMDLCSDNSIMIGWAGIIEWESLRLVSDLDICPIRQWPLNDLLSVDGWRTDQL SEQ ID NO.143, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RAD53, amino acids SequenceMENITQPTQQSTQATQRFLIEKFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSIKKVWTFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSNQLLSQGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDRIRSNLKNTSKIASPGLTSSTASSMVANKTGIFKDFSIIDEVVGQGAFATVKKAIERTTGKTFAVKIISKRKVIGNMDGVTRELEVLQKLNHPRIVRLKGFYEDTESYYMVMEFVSGGDLMDFVAAHGAVGEDAGREISRQILTAIKYIHSMGISHRDLKPDNILIEQDDPVLVKITDFGLAKVQGNGSFMKTFCGTLAYVAPEVIRGKDTSVSPDEYEERNEYSSLVDMWSMGCLVYVILTGHLPFSGSTQDQLYKQIGRGSYHEGPLKDFRISEEARDFIDSLLQVDPNNRSTAAKALNHPWIKMSPLGSQSYGDFSQISLSQSLSQQKLLENMDDAQYEFVKAQRKLQMEQQLQEQDQEDQDGKIQGFKIPAHAPIRYTQPKSIEAETREQKLLHSNNTENVKSSKKKGNGRFLTLKPLPDSIIQESLEIQQGVNPFFIGRSEDCNCKIEDNRLSRVHCFIFKKRHAVGKSMYESPAQGLDDIWYCHTGTNVSYLNNNRMIQGTKFLLQDGDEIKIIWDKNNKFVIGFKVEINDTTGLFNEGLGMLQEQRVVLKQTAEEKDLVKKLTQMMAAQRANQPSASSSSMSAKKPPVSDTNNNGNNSVLNDLVESPINANTGNILKRIHSVSLSQSQIDPSKKVKRAKLDQTSKGPENLQFS SEQ ID NO.144, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RCN1, amino acid sequence ColumnMGNIITDTIIITSDKCDIVDNDNVERIQVWLSKNILRKFQINENEPLQLIILKRFKRILLICPSHDISQHVMDASRALEMENFNFSYSLQDGQRNLTKQYLKVPESEKMFLISPPASPPPEFDFSKCEDAPQRHIQSHIQQDQQQRLEASQLLPNNPDKNNNGTFTLLKSKVGAITIDRCPTNDGNGQMQLADHVKTAFPPKSIFDTDDDD SEQ ID NO.145, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae REG1, amino acid sequence Column SEQ ID NO.146, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae REG2, amino acid sequence Column MTLSNCDSLDNLFQDPPEEEESSKFVEAVRTLMNRNDMGYPPAAANGTYCLKKIKSLNAKQWKINKKRMCMLPAVKKKNFDFHEQRSLILNLNLWKFIKFINCSSKNNYNKNNKHVRSSNNTVKNENVLPLQKHKKVDNDQRLENLFWRSWFKARKRRDIMGKPRERHIKFNDNVEQCIITDEHFIQRLPSTRLNSTDEQRPCSKSELDPCIGNAASKRSFYDYNSVYVASDAIITTAAATAIISSNSGDYQRGHDVRDVPRNVLLQAGETDFSSVLRVDSDLKLSNISHHSPVKPSSTSSHSTFIFESETDTDTDTDAETENDIDAYIDTSIPNLLL SEQ ID NO.147, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RPT1, amino acid sequence Column MPPKEDWEKYKAPLEDDDKKPDDDKIVPLTEGDIQVLKSYGAAPYAAKLKQTENDLKDIEARIKEKAGVKESDTGLAPSHLWDIMGDRQRLGEEHPLQVARCTKIIKGNGESDETTTDNNNSGNSNSNSNQQSTDADEDDEDAKYVINLKQIAKFVVGLGERVSPTDIEEGMRVGVDRSKYNIELPLPPRIDPSVTMMTVEEKPDVTYSDVGGCKDQIEKLREVVELPLLSPERFATLGIDPPKGILLYGPPGTGKTLCARAVANRTDATFIRVIGSELVQKYVGEGARMVRELFEMARTKKACIIFFDEIDAVGGARFDDGAGGDNEVQRTMLELITQLDGFDPRGNIKVMFATNRPNTLDPALLRPGRIDRKVEFSLPDLEGRANIFRIHSKSMSVERGIRWELISRLCPNSTGAELRSVCTEAGMFAIRARRKVATEKDFLKAVDKVISGYKKFSSTSRYMQYN SEQ ID NO.148, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RPT2, amino acid sequence ColumnMGQGVSSGQDKKKKKGSNQKPKYEPPVQSKFGRKKRKGGPATAEKLPNIYPSTRCKLKLLRMERIKDHLLLEEEFVSNSEILKPFEKQEEEKKQLEEIRGNPLSIGTLEEIIDDDHAIVTSPTMPDYYVSILSFVDKELLEPGCSVLLHHKTMSIVGVLQDDADPMVSVMKMDKSPTESYSDIGGLESQIQEIKESVELPLTHPELYEEMGIKPPKG VILYGAPGTGKTLLAKAVANQTSATFLRIVGSELIQKYLGDGPRLCRQIFKVAGENAPSIVFIDEIDAIGTKRYDSNSGGEREIQRTMLELLNQLDGFDDRGDVKVIMATNKIETLDPALIRPGRIDRKILFENPDLSTKKILGIHTSKMNLSEDVNLETLVTTKDDLSGADIQAMCTEAGLLALRERMRQVTAEDFKQAKERVMKNKVEENLEGLYL SEQ ID NO.149, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RPT3, amino acid sequence Column MEELGIVTPVEKAVEEKPAVKSYASLLAQLNGTVNNNSALSNVNSDIYFKLKLEKEYELLTLQEDYIKDEQRHLKRELKRAQEEVKRIQSVPLVIGQFLEPIDQNTGIVSSTTGMSYVVRILSTLDRELLKPSMSVALHRHSNALVDILPPDDSSISVMGENEKPDVTYADVGGLDMQKQEIREAVELPLVQADLYEQIGIDPPRGVLLYGPPGTGKTMLVKAVANSTKAAFIRVNGSEFVHKYLGEGPRMVRDVFRLARENAPSIIFIDEVDSIATKRFDAQTGSDREVQRILIELLTQMDGFDQSTNVKVIMATNRADTLDPALLRPGRLDRKIEFPSLRDRRERRLIFGTIASKMSLAPEADLDSLIIRNDSLSGAVIAAIMQEAGLRAVRKNRYVILQSDLEEAYATQVKTDNTVDKFDFYK SEQ ID NO.150, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RPT4, amino acid sequence ColumnMSEEQDPLLAGLGETSGDNHTQQSHEQQPEQPQETEEHHEEPSRVDPEQEAHNKALNQFKRKLLEHRRYDDQLKQRRQNIRDLEKLYDKTENDIKALQSIGQLIGEVMKELSEEKYIVKASSGPRYIVGVRNSVDRSKLKKGVRVTLDITTLTIMRILPRETDPLVYNMTSFEQGEITFDGIGGLTEQIRELREVIELPLKNPEIFQRVGIKPPKGV LLYGPPGTGKTLLAKAVAATIGANFIFSPASGIVDKYIGESARIIREMFAYAKEHEPCIIFMDEVDAIGGRRFSEGTSADREIQRTLMELLTQMDGFDNLGQTKIIMATNRPDTLDPALLRPGRLDRKVEIPLPNEAGRLEIFKIHTAKVKKTGEFDFEAAVKMSDGFGADIRNCATEAGFFAIRDDRDHINPDDLMKAVRKVAEVKKLEGTIEYQKL SEQ ID NO.151, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RPT5, amino acid sequence Column MATLEELDAQTLPGDDELDQEILNLSTQELQTRAKLLDNEIRIFRSELQRLSHENNVMLEKIKDNKEKIKNNRQLPYLVANVVEVMDMNEIEDKENSESTTQGGNVNLDNTAVGKAAVVKTSSRQTVFLPMVGLVDPDKLKPNDLVGVNKDSYLILDTLPSEFDSRVKAMEVDEKPTETYSDVGGLDKQIEELVEAIVLPMKRADKFKDMGIRAPKGALMYGPPGTGKTLLARACAAQTNATFLKLAAPQLVQMYIGEGAKLVRDAFALAKEKAPTIIFIDELDAIGTKRFDSEKSGDREVQRTMLELLNQLDGFSSDDRVKLAATNRVDVLDPALLRSGRLDRKIEFPLPSEDSRAQILQIHSRKMTDDINWQELARSTDEFNGAQLKAVTVEAGMIALRNGQSSVKHEDFVEGISEVQARKSKSVSFYA SEQ ID NO.152, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae RPT6, amino acid sequence ColumnMTAAVTSSNIVLETHESGIKPYFEQKIQETELKIRSKTENVRRLEAQRNALNDKVRFIKDELRLLQEPGSYVGEVIKIVSDKKVLVKVQPEGKYIVDVAKDINVKDLKASQRVCLRSDSYMLHKVLENKADPLVSLMMVEKVPDSTYDMVGGLTKQIKEIKEVIELPVKHPELFESLGIQPKGVILYGPPGTGKTLLARAVAHHTDCKFIRVSGAELVQKYIGEGSRMVRELFVMAREHAPSIIFMDEIDSIGSTRVEGSGGGDSEVQRTMLELLNQLDGFETSKNIKIIMATNRLDILDPALLRPGRIDRKIEFPPSVAARAEILRIHSRKMNLTRGINLRKVAEKMNGCSGADVKGVCTEAGMYALRERRRIHVTQEDFELAVGKVMNKNQETAISVAKLFK SEQ ID NO.153, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae RRD1, amino acid sequence Column MSLDRWDWPHATFSTPVKRIFDTQTTLDFQSSLAIHRIKYHLHKYTTLISHCSDPDPHATASSIAMVNGLMGVLDKLAHLIDETPLPGPRRYGNLACREWHHKLDERLPQWLQEMLPSEYHEVVPELQYYLGNSFGSSTRLDYGTGHELSFMATITALDLLGMFPHMRGADVFLLFNKYYTIMRRLILTYTLEPA GSHGVWGLDDHHFHLVYILGSSQWQLLDAQAPLQPREILDKSLVREYKDTNFYCQGINFINEVKMGPFEEHSPILYDIAVTVPRWSKVCKGLLKMYSVEVLKKFPVVQHFWFGTGFFPWVNIQNGTDLPVFEEKEEESIEQANAGSPGREQTSTRFPTSTSMPPPGVPPSGNSINYLLSHQNQSHRNQTSFSRDRLRR SEQ ID NO.154, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RRD2, amino acid sequence ColumnMLPEKRLLTPDDMKLWEESPTRAHFTKFIIDLAESVKGHENSQYKEPISESINSMMNLLSQIKDITQKHPVIKDADSSRFGKVEFRDFYDEVSRNSRKILRSEFPSLTDEQLEQLSIYLDESWGNKRRIDYGSGHELNFMCLLYGLYSYGIFNLSNDSTNLVLKVFIEYLKIMRILETKYWLEPAGSHGVWGLDDYHFLPFLFGAFQLTTHKHLKPISIHNNELVEMFAHRYLYFGCIAFINKVKSSASLRWHSPMLDDISGVKTWSKVAEGMIKMYKAEVLSKLPIMQHFYFSEFLPCPDGVSPPRGHIHDGTDKDDECNFEGHVHTTWGDCCGIKLPSAIAATEMNKKHHKPIPFD SEQ ID NO.155, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RTR1, amino acid sequence Column MATIEDIKETALIPFQKHRQLSMHEAEVITLEIIGLLCDSECKDEKTLKYLGRFLTPDMYQDLVDERNLNKRCGYPLCGKSPERIRDPFSMNDTTKKFLLENNPYAYLSHYCSKFHFRCSQFYQVQLSDEALFARTGVHLFEDPEQDKHDIDFKVTLFEELLREKASEEDIKSLISGLKKLGLNPDSGTTEKDDTELEDDLSKWLAQIKIVENDNPSILGDFTRED SEQ ID NO.156, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae RTR2, amino acid sequence Column MQIITTTFIQNVILGSHQLHEQLSIVEARMIESAIVSMLTESFCENEQTLKYLARLLSPMSYMDVINARRGKKICGYPLCYKSAAENSSDGFFIHSMYCNNYHSKCSLYLMRQLSQTPLHERRGVHLTSYINLEFDDMYSVSLLEELVGSEVPIDTVKSLITSFKDLEFDDTYKNEPLPLDVYFGQLTTDEETCIE SEQ ID NO.157, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae RTS1, amino acid sequence ColumnMMRGFKQRLIKKTTGSSSSSSSKKKDKEKEKEKSSTTSSTSKKPASASSSSHGTTHSSASSTGSKSTTEKGKQSGSVPSQGKHHSSSTSKTKTATTPSSSSSSSRSSSVSRSGSSSTKKTSSRKGQEQSKQSQQPSQSQKQGSSSSSAAIMNPTPVLTVTKDDKSTSGEDHAHPTLLGAVSAVPSSPISNASGTAVSSDVENGNSNNNNMNINTSNTQDANHASSQSIDIRRSSHSFERLPTPTKLNPDTDLELIKTPQRHSSSRFEPSRYTPLTKLPNFNEVSPEERIPLFIAKVDQCNTMFDFNDPSFDIQGKEIKRSTLDELIEFLVTNRFTYTNEMYAHVVNMFKINLFRPIPPPVNPVGDIYDPDEDEPVNELAWPHMQAVYEFFLRFVESPDFNHQIAKQYIDQDFILKLLELFDSEDIRERDCLKTTLHRIYGKFLSLRSFIRRSMNNIFLQFIYETEKFNGVAELLEILGSIINGFALPLKEEHKVFLVRILIPLHKVRCLSLYHPQLAYCIVQFLEKDPLLTEEVVMGLLRYWPKINSTKEIMFLNEIEDIFEVIEPLEFIKVEVPLFVQLAKCISSPHFQVAEKVLSYWNNEYFLNLCIENAEVILPIIFPALYELTSQLELDSANGEDSISDPYMLVEQAINSGSWNRAIHAMAFKALKIFLETNPVLYENCNALYLSSVKETQQRKVQREENWSKLEEYVKNLRINNDKDQYTIKNPELRNSFNTASENNTLNEENENDCDSEIQ SEQ ID NO.158, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SAC1, amino acid sequence ColumnMTGPIVYVQNADGIFFKLAEGKGTNDAVIHLANQDQGVRVLGAEEFPVQGEVVKIASLMGFIKLKLNRYAIIANTVEETGRFNGHVFYRVLQHSIVSTKFNSRIDSEEAEYIKLLELHLKNSTFYFSYTYDLTNSLQRNEKVGPAASWKTADERFFWNHYLTEDLRNFAHQDPRIDSFIQPVIYGYAKTVDAVLNATPIVLGLITRRSIFRAGTRYFRRGVDKDGNVGNFNETEQILLAENPESEKIHVFSFLQTRGSVPIYWAEINNLKYKPNLVLGENSLDATKKHFDQQKELYGDNYLVNLVNQKGHELPVKEGYESVVHALNDPKIHYVYFDFHHECRKMQWHRVKLLIDHLEKLGLSNEDFFHKVIDSNGNTVEIVNEQHSVVRTNCMDCLDRTNVVQSVLAQWVLQKEFESADVVATGSTWEDNAPLLTSYQNLWADNADAVSVAYSGTGALKTDFTRTGKRTRLGAFNDFLNSASRYYQNNWTDGPRQDSYDLFLGGFRPHTASIKSPFPDRRPVYIQLIPMIICAALTVLGATIFFPKDRFTSSKNLLYFAGASIVLALSTKFMFKNGIQFVNWPKLADVGFLVVHQTHDKEQQFKGLKYAQSPKFSKPDPLKRD SEQ ID NO.159, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SAP155, amino acids Sequence SEQ ID NO.160, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SAP185, amino acids Sequence SEQ ID NO.161, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SAP190, amino acids Sequence SEQ ID NO.162, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SAP4, amino acid sequence Column MSLWPFGETLSHSGIDSILEEYYLIFRSLEGNETSSTDDKKNEPSMESESEFGTESRDRSDLNQSFIDRILLETALLDELNGGANDRLVDFICLGYFYDDRSQQVRHMDYLVGMLMAYLKDIDRTGYRTPFLLENSFHQTGEYEDQDDEDPMLYVNIISSIFCSKSAPIVEALVQNTPFLSLFEVFQFENIEAENCPILAVFLKINETLLFEQTSSYLEFFKSQPNIVDKFLYHIEVSPLVEFLIKIMLTDQVESPTNIIDFLYHQDLIPKCLNLLDNSKYSPGIQNSSGELLKALISISTNFKLDTLWIGPNRLTRQLASPQYVDQLINIILFQRGHAMGVAVSIIIELIRKNNSDYDEVDLLSTTIVDNPPSQRDPVYLGHLLYELTMHMEDFYALLIKLENDDD DHDTASKALPSVKHHLLENQLHESFRPLGFERVKITELISEMLHCSNMGLMNSKRGEKIARTRDKCRDTLDQNSLEKAMKNLNINDNTITSNTLEDKCNNNDSNDSNQKQKKNIKKKFHDNELYSTFDTSDDNIDDDMSFEIPYVSETQNLKIRKNPTIGDLFKIKLHDLGFFPKFLQLFLRYPWNNFWHNIVFDIIQQIFNGRMDFSYNSFLVYSLFDFKKSTRFIPKPLYGSNQKLPVKDFHIISDFILQGHKDSFEFYEKEKTNLGYMGQLVLIAEEIAKYSKIYKTDLIAPDIYAFLQDEVWMSYSSDILNETRTMCSIILGGGQFCAESDENTNQDFLEKADMSKPAHPSTMDENEIVHEEDVKLHDKVAELIDELGQLTELDIHDKIKDVIVDHHSDLN SEQ ID NO.163, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SCD5, amino acid sequence ColumnMSFDWLNVPGLDLSSGDQAEKRPSNGLGPPSVSFDFGINTAAPHDSSFWDQGSRSHSDTTLSYRNNHSNTAADNATNVSSPQKDNPPNGEVRTLSGGDVYAESPEDMQVPLSLSQNQLTHEEIRTYLRWYHYICLRTHGKLVRLNDVFRFLTNFNLSQKVKDRIVEIFRSCKNALNIGQFFAVLRLVSRAIIYGILPLRRMILEKAPVPKPRPILSSENHEEVYEEVEDDDSSAKTGDQKVDFDSFASLLLTGKTTRKRVRRRIKNLNFKSKKVRFSEHITFQDPPNLNQESSNNSEARKQDPDAEDEDQDSNNDSPLDFTLPMDQLLKRLYKRRKNSGLVSSLPSEQQETEEEKKVLEDMKDSLSHFKQIQTVDSASLPISSVFLQNGNTLPTSNVNNTTVPQQLPLEPLKPTATGSANHLVREEYNQGLHPSNGAIQTGLQPLKPTATGSANYLMRSHMEQPQSIKPSSTPETVTNSGGLQPLKPTATGSANYLMKQHISPSVNNPVSSMFQAQFTNQSSSPQSTGPAFLNSPNITLPQSNQQQPYQEVNPTQAKIEPSNISPQHTYSNNVRINNGNIVSMPKVEITGAFPPQNTLPQHQQSHLLSPQNTIPQHQRSQLISPQNTFTQNQPILSPQHTYSNNQATMISPQNTYTNNQQQPQHLPPPPPPRAQQQQQGAIVPPQHMYSNVQKQNNLVPTQPSYTNSPSIQSPNFLSPQNAANSYFQSLLSSSPSPNPTPSNASTVNGNNASNGISSFQNTSAAMNNTQSHQTYIQQQQQQQTQQRIYGGQLSQMQQHPGQLHLNNSDIHSQPNKPNYGMLGQQVHQQQQQQQQQFPFTADVNRSNSSDILGNLQSLQQQVDALQIQYNRRP SEQ ID NO.164, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SDP1, amino acid sequence ColumnMNIYTSPTRTPNIPKSGQRPSLPMLATDERSTDKESPNEDREFVPCSSLDVRRIYPKGPLLVLPEKIYLYSEPTVKELLPFDVVINVAEEANDLRMQVPAVEYHHYRWHEDSQIALDPLSTLsIIHAATTKREKILIHCQCGLSRSATLIIAYIMKYHNLSLRHSYDLLKSRADKINPSIGLIFQLMEWEVALNAKTNVQANSYRKVP SEQ ID NO.165, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SDS22, amino acids Sequence MDKNSVNKDSEEKDERHKIEVVDDTNPDFITADSELTQDLPDDDVEVIDLVHLKIKSLEDLNLYRFKNLKQLCLRQNLIESISEVEVLPHDKIVDLDFYDNKIKHISSNVNKLTKLTSLDLSFNKIKHIKNLENLTDLENLYFVQNSISKIENLSTLKSLKNLELGGNKVHSIEPDSFEGLSNLEEIWLGKNSIPRLINLHPLKNLKILSIQSNKLKIENLEELTNLEELSHNFITKIEGLEKNLKLTTLDVTSNKITSLENLNHLSNLTDIWASFNKIDQSFESLGENLSGLSRLETIYLEGNPIQLENKTSYRRKLTMNLPPSLQKIDATYIRG SEQ ID NO.166, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SDS23, amino acids SequenceMPQNTRHTSIVEMLSTPPQLPNSTDLNSLSEQTDKNTEANKSDTESLHKSISKSSSSSSLSTLDNTEYSNNNGNSLSTLNSQNLLSVHRQEWQHTPLSNLVEQNKLIFIRGSISVEEAFNTLVKHQLTSLPVENFPGDMNCLTFDYNDLNAYLLLVLNRIKVSNDKITSDCQNGKSVPVGEIVKLTPKNPFYKLPETENLSTVIGILGSGVHRVAITNVEMTQIKGILSQRRLIKYLWENARSFPNLKPLLDSSLEELNIGVLNAARDKPTFKQSRVISIQGDEHLIMALHKMYVERISSIAVVDPQGNLIGNISVTDVKHVTRTSQYPLLHNTCRHFVSVILNLRGLETGKDSFPIFHVYPTSSLARTFAKLVATKSHRLWIVQPNDNQPTASSEKSSSPSPSTPPVTTLPSLASSYHSNTQSSRMANSPVLKSSDTSNNKINVNINLSGPSPSQPQSPSATMPPPQSPSNCPASPTPAHFEKEYRTGKLIGVVSLTDILSVLARKQTHHKEIDPQMARKQRGHIG SEQ ID NO.167, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SDS24, amino acids SequenceMASTSNTFPPSQSNSSNNLPTSRHASIVEMLSTPPLLPHVQVNDTDDKEQPEESTPPTATAAAPGPGSAATPAPLRDEKPQFKLSAVPMTQTPSQCLSCVHAQKWQHIPLSQLIEQNKLIFVPGSISVEEAFNTLIKYHLNSIPVESFPGDMNCFTFDYNDLNSYLLLVLNKITVSNKQLTADCQNGKPVPVGEMVKLTPKNSFYKLPENESLSTVMGILGSGVHRVAITNEEMTKVKGILSQRRLIKYLWDNARSFTSLEPLLNSSLQDLHIGVLNIQSKPTSRQSRVISIQGEEPLIMGLYKMHVERISSIAVIDKQGNLLGNISVTDVKHVTRTSQYPLLHKTCRHFISVILNSRGLETGKDSFPIFHVYPSSSLARTLAKLVATKSHRLWIVQPPESSTSASSTNLTAANTAANAVSATAQSSANGATPMSKSSSSTSLNSHSPLMTAMEDPPSPRSSAIAIPPPSPASSTNTPNLFEKEYRTGKLIGVVSLTDIINLLARKQTGNKEVDPQSARRQRGSIAM SEQ ID NO.168, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SDT1, amino acid sequence Column MTVEYTASDLATYQNEVNEQIAKNKAHLESLTHPGSKVTFPIDQDISATPQNPNLKVFFFDIDNCLYKSSTRIHDLMQQSILRFFQTHLKLSPEDAHVLNNSYYKEYGLAIRGLVMFHKVNALEYNRLVDDSLPLQDILKPDIPLRNMLLRLRQSGKIDKLWLFTNAYKNHAIRCLRLLGIADLFDGLTYCDYSRTDTLVCKPHVKSFEKAMKESGLARYENAYFIDDSGKNIETGIKLGMKTCIHLVENEVNEILGQTPEGAIVISDILELPHVVPDLF SEQ ID NO.169, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SER2, amino acid sequence ColumnMSKFVITCIAHGENLPKETIDQIAKEITESSAKDVSINGTKKLSARATDIFIEVAGSIVQKDLKNKLTNVIDSHNDVDVIVSVDNEYRQAKKLFVFDMDSTLIYQEVIELIAAYAGVEEQVHEITERAMNNELDFKESLREVKLLQGLQVDTLYDEIKQKLEVTKGVPELCKFLHKKNCKLAVLSGGFIQFAGFIKDQLGLDFCKANLEVDTDGKLTGKTLGPIVDGQCKSETLLQLCNDYNVPVEASCMVGDGGNDLPAMATAGFGIAWNAKPKVQKAAPCKLNTKSMTDILYILGYTDDEIYNRQ SEQ ID NO.170, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SHB17, amino acids Sequence MPSLTPRCIIVRHGQTEWSKSGQYTGLTDLPLTPYGEGQMLRTGESVFRNNQFLNPDNITYIFTSPRLRARQTVDLVLKPLSDEQRAKIRVVVDDDLREWEYGDYEGMLTREIIELRKSRGLDKERPWNIWRDGC ENGETTQQIGLRLSRAIARIQNLHRKHQSEGRASDIMVFAHGHALRYFAAIWFGLGVQKKCETIEEIQNVKSYDDDTPYVKLESYRHLVDNPCFLLDAGGIGVLSYAHNIDEPALELAGPFVSPPEEESQHGDV SEQ ID NO.171, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SHP1, amino acid sequence ColumnMAEIPDETIQQFMALTNVSHNIAVQYLSEFGDLNEALNSYYASQTDDQKDRREEAHWNRQQEKALKQEAFSSTNSNKAINTEHVGGLCPKPGSSQGSNEYLKRKGSTSPEPTKGSSRSGSNNSRFMSFSDMVRGQADDDDEDQPRNTFAGGETSGLEVTDPSDPNSLLKDLLEKARRGGQMGAENGSRDDEDQEMGANRFTGRGFRLGSTIDAADEVVEDNTSQSQRRPEKVTREITFWKEGFQVADGPLYRYDDPANSFYLSELNQGRAPLKLLDVQFGQEVEVNVYKKLDESYKAPKRKLGGFSGQQQRLGSPIPGESSPAEVPKNETPAAQEQPMPNNEPKQGDTSIQIRYANGKREVLRCNSTDTVKFLYEHVTSNANTDPSRNFTLNYAFPIKPISNDETTLKDADLLNSVVQRWA SEQ ID NO.172, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SIA1, amino acid sequence ColumnMRLHYRRRFNFLRRILFILCITSLYLSRDSLKLHAKNVLMDHNVAEYHGGMIDDIQILRCYHWYRQCSSLYAPKLHPSNTAKKIKDKNSILWTRVSKNITVETLYSLQSGPFYNSYLYVHLKDFQSNPKNTIKELAIARDSALIPLQVLRDINKLVKSSDSSVFHNHVYLREKPTSSWWKLLFGISVDTDNIAVFGEEWVYKGSGIWCKYILNDDDNDAPITNLEIYLGSSFIESRPSWKEVIHEFHRNNIPSLPISITRKLETKNHHHKFSNGLLGSLRTPSKDINIQVDADYKITSPHIQFSRGQRSFKILQITDFHFKCTDNSMTVINEIKTVNFIDRVLASENPDLVVITGDLLDSHNTIDYQTCIMKVVQPMISNKIPYAISLGVSDESNLATSAQIRDFIRNLPYTFNNVASEEGHMAIEVSFKKKLTKNTLLERDIDTEDETNPSEALFFVFDSFAPVNNFLQDYNDLIGKIDFGLAFQYFPLSEYRPHGLFPIIGQYNERSTLTVDTPRSRGQVSMTINGKHYKSFLDILSLWNIKGVSCGHEHNNDCCLQSKNEMWLCYGGSAGIGLPRIQGIYPTVRLFNLDDILDEITSWKRNSNLVDEVYDYQYIYKGKQ SEQ ID NO.173, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SIS2, amino acid sequence ColumnMTAVASTSGKQDADHNQSIECPRFSRGQKEILLDHEDAKGKDSIINSPVSGRQSISPTLSNATTTTTKSIMNATGTSGAVVSNTPEPGLKRVPAVTFSDLKQQQKQDSLTQLKNDSERTKSPNSNPAPVSNSIPGNHAVIPNHTNTSRTTQLSGSPLVNEMKDYDPKKKDSALKIVDTMKPDKIMATSTPISRENNKVTAKAPTSITLRKEDAQDQANNVSGQINVRSTPEETPVKQSVIPSIIPKRENSKNLDPRLPQDDGKLHVLFGATGSLSVFKIKPMIKKLEEIYGRDRISIQVILTQSATQFFEQRYTKKIIKSSEKLNKMSQYESTPATPVTPTPGQCNMAQVVELPPHIQLWTDQDEWDAWKQRTDPVLHIELRRWADILVVAPLTANTLSKIALGLCDNLLTSVIRAWNPSYPILLAPSMVSSTFNSMMTKKQLQTIKEEMSWVTVFKPSEKVMDINGDIGLGGMMDWNEIVNKIVMKLGGYPKNNEEEDDDEDEEEDDDEEEDTEDKNENNNDDDDDDDDDDDDDDDDDDDDEDEDEAETPGIIDKHQ SEQ ID NO.174, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SIT4, amino acid sequence Column MVSRGPDEWLETIKKCQALTENEMKQLCEMVKELLMEESNIQPVQTPVTVCGDIHGQFHDLLELFRTAGGFPDDINYIFLGDYVDRGYYSLETFTLLMCLKVKYPAKITLVRGNHESRQITQVYGFYEECLNKYGSTTVWKYCCQVFDFLTLAAIIDGKILCVHGGLSPEIRMLDQIRVLSRAQEVPHEGGFSDLLWSDPDNVEAWQVSPRGAGWLFGSKVAREFNHVNGLNLIARAHQLVMEGFKYHFPEKDVVTVWSAPNYCYRCGNVASVMKVDEDLEPTFKIFSAVPDDYIRESTANHNNQRAGYFL SEQ ID NO.175, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae SIW14, amino acids SequenceMGLYQAKNDEGSDPKSSSKIDDLIENEAEIIRLIKEDGKLLIDNGDGRDIHNIIQEDKLLSVEFNEVLKRFHGEEKSDIPRKEFDEDEDDGYDSNEHHQKTIEVMNTLNHVINKEVIPPENFSHVVGEIYRSSFPRQENFSFLHERLKLKSILVLIPEEYPQENLNFLKLTGIKLYQVGMSGNKEPFVNIPSHLLTKALEIVLNPANQPILIHCNRGKHRTGCLIGCIRKLQNWSLTMIFDEYRRFAFPKARALDQQFIEMYDDDEIKRIASKNNWLPLQW SEQ ID NO.176, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SMN1, amino acid sequence Column MILKLVHCLVALTGLIFAKPYQQQQAVLAPSQDVPLRDIHIGDINFIHTTDTHGWLGSHLSQNDYDADWGDFVAFVDILREKILRQSRDVIVIDTGDKRDGNGLSDATWPPGLRSSEIFNMMDYDLLTLGNHELYTAESAILEYRGTSQSSKFKDKYVCSNVEFIEDDGTRVPFGNKYITFETPIMKQRVLALSFLFSFQRANNRAIVTPPLEEITQKSWFQNMVETNREEEIDLIIVFGHLPATDPTEREMHKIHALIRKYYPNTVIQYFGGHTHIRDFVQLDSKSTCLQSGRFAETVGFLSINMTDPVDAESPIFSRRYIDFNKEAFKYHLSKLGHDSNVPVSTKKGKTISRLVNDLRHELNLNEKLGYIPQTYYVSTRPLNSEENLYHLITHKILPNLIPPKNYEPSMSRFILINTGSVRYDLYKGPFTKDTEYIVMPFNNDWRFITVPLVVASRVETYLNKGPVIASLGIPSSSHHKQHFGGFQKCPFINNPNLSEGYTTEDDFGCHGDDTPHNSQREYDIPNVVQCKEVKKVQEEEADPSKMVHVIFYSFMELDILNAVNSIINDLGLRMENLTTNDCSHYGGDSTKKLLRDYFSQF SEQ ID NO.177, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SpO7, amino acid sequence ColumnMEPESIGDVGNHAQDDSASIVSGPRRRSTSKTSSAKNIRNSSNISPASMIFRNLLLILEDDLRRQAHEQKILKWQFTLFLASMAGVGAFTFYELYFTSDYVKGFHRVILQFTLSFISITVVLFHISGQYRRTIVIPRRFFTSTNKIRQFNVKLVKVQSTWDEKYTDSVRFVSRTIAYCNIYCLKKFLWLKDDNAIVKFWKSVTIQSQPRIGAVDVKLVLNPRAFSAEIREGWEIYRDEFWAREGARRRKQAHELRPKSE SEQ ID NO.178, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae SSU72, amino acids Sequence MPSHRNSNLKFCTVCASNNNRSMESHKVLQEAGYNVSSYGTGSAVRLPGLSIDKPNVYSFGTPYNDIYNDLLSQSADRYKSNGLLQMLDRNRRLKKAPEKWQESTKVFDFVFTCEERCFDAVCEDLMNRGGKLNKIVHVINVDIKDDDENAKIGSKAILELADMLNDKIEQCEKDIPFEDCIMDILTEWQSSHSQLPSLYAPSYY SEQ ID NO.179, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae STT4, amino acid sequence Column SEQ ID NO.180, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TAP42, amino acids Sequence MASVTEQFNDIISLYSTKLEHTSLRQDSPEYQGLLSTIKKLLNLKTAIFDRLALFSTNETIDDVSTASIKFLAVDYYLGLLISRRQSNDSDVAQRQSMKLIYLKKSVESFINFLTLLQDYKLLDPLVGEKLGNFKDRYNPQLSELYAQPKNNKDLSGAQLKRKKEIELFQRNKEISTKLHCLELELKNNDEDHDHDELLRELYLMRLHHFSLDTINNIEQNLFECEMLSNFLKNSVHEVKSSGTQIRKESNDDDSTGFTDKLENINKPLIDKKQVLRNFTLVDKRQQLQQKVRGYGQYGPTMSVEEFLDKEFEEGRVLQGGEEPEQAPDEENMDWQDRETYKAREWDEFKESHAKGSGNTMNRG SEQ ID NO.181, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TEP1, amino acid sequence Column MREEGSELEMEKGFLKWKPVNLMKKILSLPMKKTKKNDIGLRLDISYILVNLIVCSYPVNTYPKLLYRNSLDDLILFLTVYHGKGNFRIFNFRGEKEDSDYKDNDLIGIAAKFESKDFEIQELRSTLINDGKIPISPIDLETRTLVEEEETNNVICERIGWLDHFPPPFELLEEIVDGIENYLSVSKNRVAVLHCRMGKGRSGMITVAYLMKYLQCPLGEARLIFMQARFKYGMTNGVTIPSQLRYLRYHEFFITHEKAAQEGISNEAVKFKFKFRLAKMTFLRPSSLITSESAIVTTKIQHYNDDRNALLTRKVVYSDIMAHECGGNMTFIFGRDYLTLENDCRIEFTLGTSKSKAASSIISWTSCASCWLNIYLETLMHIIKDDSSPDYFQVERLKRDEMLGTTISWQELDGFGELSTHGLKLFQALKLEWEII SEQ ID NO.182, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae TFC7, amino acid sequence ColumnMVVNTIYIARHGYRSNWLPEGPYPDPLTGIDSDVPLAEHGVQQAKELAHYLLSLDNQPEAAFASPFYRCLETVQPIAKLLEIPVYLERGIGEWYRPDRKPVIPVPAGYEILSKFFPGVISQEWDSTLTPNEKGETEQEMYMRFKKFWPLFIERVEKEYPNVECILLVTHAASKIALGMSLLGYDNPRMSLNENGDKIRSGSCSLDKYEILKKSYDTIDETDDQTSFTYIPFSDRKWVLTMNGNTEFLSSGEEMNWNFDCVAEAGSDADIKKRQMTKKTSSPIPEADDQTEVETVYISVDIPSGNYKERTEIAKSAILQYSGLETDAPLFRIGNRLYEGSWERLVGTELAFPNAAHVHKKTAGLLSPTEENETTNAGQSKGSSTANDPNIQIQEEDVGLPDSTNTSRDHTGDKEEVQSEKIYRIKERIVLSNVRPM SEQ ID NO.183, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TFG1, amino acid sequence ColumnMSRRNPPGSRNGGGPTNASPFIKRDRMRRNFLRMRMGQNGSNSSSPGVPNGDNSRGSLVKKDDPEYAEEREKMLLQIGVEADAGRSNVKVKDEDPNEYNEFPLRAIPKEDLENMRTHLLKFQSKKKINPVTDFHLPVRLHRKDTRNLQFQLTRAEIVQRQKEISEYKKKAEQERSTPNSGGMNKSGTVSLNNTVKDGSQTPTVDSVTKDNTANGVNSSIPTVTGSSVPPASPTTVSAVESNGLSNGSTSAANGLDGNASTANLANGRPLVTKLEDAGPAEDPTKVGMVKYDGKEVTNEPEFEEGTMDPLADVAPDGGGRAKRGNLRRKTRQLKVLDENAKKLRFEYPWVMEDFDGYNTWVGSYEA GNSDSYVLLSVEDDGSFTMIPADKVYKFTARNKYATLTIDEAEKRMDKKSGEVPRWLMKHLDNIGTTTTRYDRTRRKLKAVADQQAMDEDDRDDNSEVELDYDEEFADDEEAPIIDGNEQENKESEQRIKKEMLQANAMGLRDEEAPSENEEDELFGEKKIDGERIKKALQKTELAALYSSD ENEIPYLSDIENKENESPVKKEEDSDTLSKSKRSSPKKQQKKATNAHVHKEPTLRVKSIKNCVIILKGDKKILKSFPEGEWNPQTTKAVDSSNNASNSTVPSPIKQEEGLNSTVAEREETPAPTITEKDIIEAIGDGKVNIKEFGKFIRRKYPGAENKKLMFAIVKKLCRKVGNDHMELKKE SEQ ID NO.184, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae TOF2, amino acid sequence ColumnMIKMWRLQIVLVPPSAQDIITFLEARLNTPQSVSPMVQYNEDIITHNNSINNCSDSPSTPSPSSQNSIQSNRSSDFINYLPNCCKKFLHFTDGDNTLLQLSNEILTKFDRLYPNFKESIEIVSLQDRHGCDLDSEFIIKDVFENDGVVLVILKDELDWSRNQHISLLQLARQRRRQDNKPSTKSIVTEKRKKISKEDLSSISNKDTMHLIAKSSLKNNFINKSRVSTPLMNEILPLASKYDALNKEKCPMPLTSTVVASNVHKDVKDHARAKEGVVTQGSDNNKENIPSSTQQQKNDGAKRAESKDLLLRNSSEDADYEPADENSPQISFDSIDTDFQLSTTSHTNSDMHIQYSNPSSGAHSPRKSSLEIKVQNKKGDDLPLNDKD IGENCRRIEAFSDEEDFNETDNDRADSFINNSKKASMGFRDINSDLDSVSFNSDIENAVQSTQSTKNVVSPPFFPEKELNNRLHQSQGKEALFRLVEKEFPDKSLGAASSTSHAKDVKIQETIRKLNRFKPTGETKVQKRNSITEPYYGKFGIMKKDKPKSITSKGVSLETKHFDDPNTIISGEKFAKFGKIKVKRKTDDVGSKVIEFKRKRNMGNRSLKDIFANAGKPPNAASTIKVVKLMRDPVDNSKDKVEATSNSTAQEQEQVSPKLPVMNSTPGKRKNGQAIPSSLERTPQLKKVKVTRSHSSPSSSSSMSLESSLDSSSSDDSDDDSRNVQVKKINFKTSHGPAGNSNGKPMLDVDDNEINTKKYQTPKYVESDEDDQ SEQ ID NO.185, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TPD3, amino acid sequence ColumnMSGARSTTAGAVPSAATTSTTSTTSNSKDSDSNESLYPLALLMDELKHDDIANRVEAMKKLDTIALALGPERTRNELIPFLTEVAQDDEDEVFAVLAEQLGKFVPYIGGPQYATILLPVLEILASAEETLVREKAVDSLNNVAQELSQEQLFSDFVPLIEHLATADWFSSKVSACGLFKSVIVRIKDDSLRKNILALYLQLAQDDTPMVKRAVGKNLPILIDLLTQNLGLSTDEDWDYISNIFQKIINDNQDSVKFLAVDCLISILKFFNAKGDESHTQDLLNSAVKLIGDEAWRVRYMAADRFSDLASQFSSNQAYIDELVQPFLNLCEDNEGDVREAVAKQVSGFAKFLNDPSIILNKILPAVQNLSMDESETVRSALASKITNIVLLLNKDQVINNFLPILLNMLRDEFPDVRLNIIASLKVVNDVIGIELLSDSLLPAITELAKDVNWRVRMAIIEYIPILAEQLGMQFFDQQLSDLCLSWLWDTVYSIREAAVNNLKRLTEIFGSDWCRDEIISRLLKFDLQLLENFVSRFTILSALTTLVPVVSLDVVTEQLLPFISHLADDGVPNIRFNVAKSYAVIVKVLIKDEAKYDALIKNTILPSLQTLCQDEDVDVKYFAKKSLAECQELLKN SEQ ID NO.186, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae TPP1, amino acid sequence List MSHKLTILPFLIKFTPKFPQSIDHDEHGLNVYAFDLDHTIIKPKSPNISFSRSASDWQFINFNSKKSTLDYLCNIIDNDPTAVIVIFSNQGGVITVPRTSKSCTKYTNKILLFLKAIKNDERGETLSHRLWLYAAPKRPKTFAANHSKITFASLGESYNNDPNIFEKVRKPMTGMVEFFKRDLESAYRVSEQISPIKLNWIYYCGDAAGRKKDFSDSDIKFAENLHVEFKYPEEIFHG SEQ ID NO.187, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TPS1, amino acid sequence ListMTTDNAKAQLTSSSGGNIIVVSNRLPVTITKNSSTGQYEYAMSSGGLVTALEGLKKTYTFKWFGWPGLEIPDDEKDQVRKDLLEKFNAVPIFLSDEIADLHYNGFSNSILWPLFHYHPGEINFDENAWLAYNEANQTFTNEIAKTMNHNDLIWVHDYHLMLVPEMLRVKIHEKQLQNVKVGWFLHTPFPSSEIYRILPVRQEILKGVLSCDLVGFHTYDYARHFLSSVQRVLNVNTLPNGVEYQGRFVNVGAFPIGIDVDKFTDGLKKESVQKRIQQLKETFKGCKIIVGVDRLDYIKGVPQKLHAMEVFLNEHPEWRGKVVLVQVAVPSRGDVEEYQYLRSVVNELVGRINGQFGTVEFVPIHFMHKSIPFEELISLYAVSDVCLVSSTRDGMNLVSYEYIACQEEKKGSLILSEFTGAAQSLNGAIIVNPWNTDDLSDAINEALTLPDVKKEVNWEKLYKYISKYTSAFWGENFVHELYSTSSSSTSSSATKN SEQ ID NO.188, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae TPS2, amino acid sequence ListMTTTAQDNSPKKRQRIINCVTQLPYKIQLGESNDDWKISATTGNSALFSSLEYLQFDSTEYEQHVVGWTGEITRTERNLFTREAKEKPQDLDDDPLYLTKEQINGLTTTLQDHMKSDKEAKTDTTQTAPVTNNVHPVWLLRKNQSRWRNYAEKVIWPTFHYILNPSNEGEQEKNWWYDYVKFNEAYAQKIGEVYRKGDIIWIHDYYLLLLPQLLRMKFNDESII IGYFHHAPWPSNEYFRCLPRRKQILDGLVGANRICFQNESFSRHFVSSCKRLLDATAKKSKNSSNSDQYQVSVYGGDVLVDSLPIGVNTTQILKDAFTKDIDSKVLSIKQAYQNKKIIIGRDRLDSVRGVVQKLRAFETFLAMYPEWRDQVVLIQVSSPTANRNSPQTIRLEQQVNELVNSINSEYGNLFSPVQHYYMRIPKDVYLSLLRVADLCLITSVRDG MNTTALEYVTVKSHMSNFLCYGNPLILSEFSGSSNVLKDAIVVNPWDSVAVAKSINMALKLDKEEKSNLESKLWKEVPTIQDWTNKFLSSLKEQASSNDDMERKMTPALNRPVLLENYKQAKRRLFLDYDGTLTPIVKDPAAAIPSARLYTILQKLCADPHNQIWIISGRDQKFLNKWLGGKLPQLGLSAEHGCFMKDVSCQDWVNLTEKVDMSWQVRVNEVMEEFTTRTPGSFIERKKVALTWHYRRTVPELGEFHAKELKEKLLSFTDDFDLEVMDGKANIEVRPRFVNKGEIVKRLVWHQHGKPQDMLKGISEKLPKDEMPDFVLCLGDDFTDEDMFRQLNTIETCWKEKYPDQKNQWGNYGFYPVTVGSAKTVAKAHLTDPQQVLETLGLLVGDVSLFQSAGTVDLDSRGHVKNSESSLKSKLASKAYVMKRSASYTGAKV SEQ ID NO.189, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae TPS3, amino acid sequence List SEQ ID NO.190, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae TSL1, amino acid sequence List SEQ ID NO.191, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae UTR4, amino acid sequence List MGDNYSTYLLDIEGTVCPISFVKETLSPYFTKKVPQLVQQDTRDSPVSNILSQFHIDDKEQLQAHILELVAKDVKDPILKQLQGYIWAQGYESGQIKAPVYADAIDFIRKKRVFIYSSGSVKAQKLLFGYVQDPNAPAHDSLDLNSYIDGYFDINTSGKKTETQSYANILRDIGAKASEVLFLSDNPLELDAAAGVGIATGLASRPGNAPVPDGQKYQVYKDFETL SEQ ID NO.192, derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae VHS3, amino acid sequence List MTNKSSLKNNRKGVASNTLSGAEQANIGSSAMPDTNSTGPFSSVSSLDTPVVRKSTSPTGSQTKSIMNASGTSGAVVSNTPEPGLKRIPTVTFSDPKLGSLRSDVEQTPPNQVARQSSEKKATSVHIAAEGANQGRNLKDINTKVPKDGEASASSFSTPTSILSNADMGNNISSLAKKLSFTGGTDSILNSDNSSDSPRKEHPHFYVEDPLHTPSVRSRSNSTSPRPSVVVNTFNPINIEREGSISKTGEPTLLESVLEEAMSPNAVSNPLKRENIMTNMDPRLPQDDGKLHVLFGATGSLSVFKLKHMIRKLEEIYGRDKICIQVILTNSATKFF AMKYMRKNKKQHNSIDTSFNSTNSNAGNITGNKKKVASLEKFSIQKTSSNSAASQTNNKQEEEKQMASTTGFPSTLGGSRTYSNSSNVVSQHPQIELPAHIQFWTDQDEWDVWRQRTDPVLHIELRRWADILVVAPLTANTLAKIALGLCDNLLTSVIRAWNPTFPIF LAPSMGSGTFNSIMTKKHFRIIQEEMPWVTVFKPSEKVMGINGDIGLSGMMDANEIVGKIVVKLGGYPVSAKGEEEEDEDNDEEDDNKKNDTGGKDEDNDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDEDEDEDDEGGKKKEDKGGLQRS SEQ ID NO.193, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae VIP1, amino acid sequence List SEQ ID NO.194, derived from Saccharomyces cerevisiae (Saccharomyces cer Saccharomyces cerevisiae VPS34, amino acids sequence MSLNNITFCVSQDLDVPLKVKIKSLEGHKPLLKPSQKILNPELMLIGSNVFPSDLIVSLQVFDKERNRNLTPYIPFRNSRTWDYWLTLPIRIKQLTFSSHLRIILWEYNGSKQIPFFNLETSIFNLKDCTLKRGFESLKFRYDVIDHCEVVTDNKDQENLNKYFQGEFTRLPWLDEITISKLRKQRENRTWPQGTFVLNLEFPMLELPVVFIEREINMNTQMNIPTLKNNPGLSTDLREPNRNDPQIKISLGDKYHSTLKFYDPDQPNNDPIEEKYRRLERASKNANLDKQVKPDIKKRDYLNKIINYPPGTKLTAHEKGSIWKYRYYLMNNKKALTKLLQSTNLREESERVEVLELMDSWAEIDIDDALELLGSTFKNLSVRSYA...

Claims

1. A method for producing perillaldehyde and / or perillyl lactone, comprising: The following components are contacted: component (1) and component (2) are brought together to produce perillaldehyde and / or perillyl lactone; wherein, The first component is selected from components containing lysine diol. The second component is selected from enzymes or microorganisms or their cell lysates or enzyme extracts that can convert lysine diol into perillyl diol and / or perillyl lactone.

2. The method of claim 1, wherein the microorganism capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone is selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganism capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone is selected from... Hyphozyma roseoniger Or Cryptococcus lightensis ( Cryptococcus albidus ).

3. The method of claim 1 or 2, wherein the microorganism capable of converting lysine-1-benzenediol into perillylenediol and / or perillylene lactone is selected from fungi with accession number ATCC 20624, fungi with accession number ATCC 20918, or derivatives thereof.

4. The method according to any one of claims 1-3, wherein the component containing lysinediol comprises isolated lysinediol, a mixture containing lysinediol, a microorganism capable of producing lysinediol or its culture medium, supernatant, cell lysate or extract.

5. The method according to any one of claims 1-4, wherein the component comprising lysinediol is a microorganism capable of producing lysinediol or its culture medium, supernatant, cell lysate or extract.

6. The method according to any one of claims 1-5, wherein the method is used to produce perillaldehyde, or to produce perillaldehyde diol and perillaldehyde.

7. The method according to any one of claims 1-5, wherein the method is used to produce perillaldehyde, and the microorganism capable of converting lysine-3-diol into perillaldehyde and / or perillaldehyde lactone is selected from fungi or derivatives thereof with accession number ATCC 20918.

8. The method according to any one of claims 1-7, wherein the microorganism capable of producing lysine diol is a recombinant engineered bacterium capable of producing lysine diol.

9. The method of claim 8, wherein, The recombinant engineered bacteria capable of producing lysine-6-ol contains a phosphatase that catalyzes the production of lysine-6-ol pyrophosphate (LPP) to lysine-6-ol (LOH). This phosphatase is derived from Mg(2+)-dependent phosphatidate phosphatase, phosphatidic acid phosphatase type 2, serine / threonine-protein phosphatase, HAD-like hydrolase superfamily, phosphate (PA) phosphatase, polyphosphoinositide phosphatase, serine / threonine-protein kinases, alkaline phosphatase, and phosphoprotein phosphatase family. This includes the following families of phosphatases: serine / threonine-protein phosphatases, SIT4 phosphatase-associated protein family, TAP42 / TAP46-like superfamily, dual-specificity lipid and protein phosphatases, polynucleotide kinase 3 phosphatases, 5'-deoxynucleotidases, myosin family, histidine phosphatase superfamily, S-2-haloalkanoic acid dehalogenase, phosphatidylglycerol phosphatase, and HAD-hydrolase superfamily. ), phosphatase family (PA-phosphatase)Related phosphoesterase family, squalene / phytoene synthase family, terpene cyclase family, phosphatase App1, protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), halogen dehalogenase (HAD), sugar phosphate phosphatase (HAD-like), polyphosphoinositide phosphatase (Fig4-like), casein kinase 1 (Serine / Threonine protein kinase), lipoprotein family, PPZ / Ppq1 family, clade-1 family, HAD-IA family hydrolases (HAD... The enzymes are preferably IAhydrolase family, HAD-IF subfamily (IF family), dolichoyldiphosphatase family, or haloperoxidase superfamily; preferably, the phosphatase is phosphatidylate phosphatase (APP1), dolichoyldiphosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), inositol polyphosphate phosphatase (FIG4), 1-glycerophosphate phosphatase 1 (GPP1), casein kinase I (HRR25), phosphatidylate phosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), or serine / threonine protein phosphatase (PTC3). Serine / threonine protein phosphatase (PTC3), SIT4-associated protein SAP155, SIT4-associated protein SAP185, type 2A phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate phosphatase (TEP1), polynucleotides3'-phosphatase (TPP1), phosphorylhydrolase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphorylhydrolase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), phosphatidylglycerol phosphatase (PgpA), phosphatidylglycerol phosphatase (PgpB), phosphatidylglycerol phosphatase (PgpC), carbapenyl diphosphatase (YbjG), dihydroxy diphosphatase (DOLPP1), phosphorylhydrolase (PLPP6), farnesyl diphosphatase (YisP), farnesol synthase (TPS2), acyclic sesquiterpene synthase (TPS1), or farnesol synthase (TPS13).

10. The method of claim 9, wherein, The phosphate hydrolase is a phosphate hydrolase derived from microorganisms or its functional variants, including bacteria, yeast, or fungi, wherein the bacteria are selected from Escherichia or Bacillus, the yeast is selected from Saccharomyces, or the phosphate hydrolase is derived from plants or humans. Homo sapiens Phosphohydrolases or their functional variants thereof; preferably, the phosphohydrolase is a phosphohydrolase or its functional variant derived from bacteria, yeast or fungi, wherein the bacteria are selected from *Escherichia coli* (…). Escherichia coli Bacillus subtilis ( Bacillus subtilis The yeast is brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), or the phosphorylase is derived from plants or humans ( Homo sapiens Phosphorylase or a functional variant thereof, wherein the plant is selected from moso bamboo ( Phyllostachys edulis ),corn( Zea mays ) or japonica rice ( Oryza sativa subsp. japonica More preferably, the phosphorylase is derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), Escherichia coli ( Escherichia coli ), human beings ( Homo sapiens Bacillus subtilis ( Bacillus subtilis ),bamboo( Phyllostachys edulis ),corn( Zea mays ) or japonica rice ( Oryza sativa subsp. japonica Phosphohydrolases or their functional variants.

11. The method of claim 9 or 10, wherein the phosphatase comprises an amino acid sequence selected from SEQ ID NO. 11-226, or an amino acid sequence having at least 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% sequence identity with an amino acid sequence selected from SEQ ID NO. 12, SEQ ID NO. 17, SEQ ID NO. 20, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 45, SEQ ID NO. 58, SEQ ID NO. 63, SEQ ID NO. 14, SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 19 ... ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID The amino acid sequence of NO.225, SEQ ID NO.226, or selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ IDNO.159, SEQ ID NO.

160. SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID The amino acid sequence of NO. 226 has at least 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% sequence identity.

12. The method according to any one of claims 9-11, wherein the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 237-452, or by a nucleotide sequence having at least 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% sequence identity with a nucleotide sequence selected from SEQ ID NO. 238, SEQ ID NO. 243, SEQ ID NO. 246, SEQ ID NO. 262, SEQ ID NO. 264, SEQ ID NO. 271, SEQ ID NO. 28, SEQ ID NO. 29, SEQ ID NO. 20 ... NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID The nucleotide sequence of NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ ID NO.452, or selected from the group consisting of SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO.271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ IDNO.359, SEQ ID NO.

383. SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ IDNO. 452 encodes a nucleotide sequence with at least 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% sequence identity.

13. The method as described in any one of claims 9-12, wherein, The recombinant engineered bacteria capable of producing lysine diol also contains genes for the lysine diol pyrophosphate (LPP) synthesis pathway.

14. The method of claim 13, wherein the lysine pyrophosphate (LPP) synthesis pathway gene includes the farnesyl pyrophosphate (FPP) synthesis pathway gene, the geranylgeranyl pyrophosphate synthase (GGPPS) gene, and the lysine pyrophosphate synthase (LPPS) gene.

15. The method of claim 14, wherein the farnesyl pyrophosphate (FPP) synthesis pathway gene comprises one or more of the following: acetyl-CoA transferase gene, 3-methyl-3-hydroxyglutaryl-CoA synthase gene, 3-hydroxy-3-methylglutaryl-CoA reductase gene, mevalonate kinase gene, mevalonate-5-phosphate kinase gene, mevalonate-5-pyrophosphate decarboxylase gene, pentene pyrophosphate isomerase gene, or farnesyl pyrophosphate synthase gene.

16. The method according to any one of claims 9-15, wherein, The recombinant engineered bacteria capable of producing lysine diol further comprises one or more of the following gene modifications: (1) At least one gene modification that enhances the activity of geraniol geraniol pyrophosphate synthase (GGPPS); and (2) At least one gene modification that enhances the activity of lysine pyrophosphate diol ester synthase (LPPS).

17. The method of claim 16, wherein, At least one gene modification that enhances the activity of gerany-gerany-gerany pyrophosphate synthase (GGPPS) includes: introducing an exogenous gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, increasing the gene copy number of the gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, and / or replacing the natural promoter of the endogenous gerany-gerany pyrophosphate synthase (GGPPS) with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of lysandrindiol pyrophosphate synthase (LPPS) includes: introducing an exogenous lysandrindiol pyrophosphate synthase (LPPS) gene, increasing the gene copy number of the lysandrindiol pyrophosphate synthase (LPPS) gene, and / or replacing the natural promoter of the endogenous lysandrindiol pyrophosphate synthase (LPPS) with a promoter that has a higher expression level.

18. The method of claim 17, wherein, The exogenous gerany-gerany pyrophosphate synthase (GGPPS) comprises the amino acid sequence shown in SEQ ID NO. 9, preferably encoded by the nucleotide sequence of SEQ ID NO. 235; and / or The exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a truncated lysine pyrophosphate diol ester synthase (LPPS); preferably, the truncated lysine pyrophosphate diol ester synthase (LPPS) is a signal peptide with the first 63 amino acids removed; preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) comprises the amino acid sequence shown in SEQ ID NO. 10, more preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) is encoded by the nucleotide sequence of SEQ ID NO.

236.

19. The method according to any one of claims 9-18, wherein, The recombinant engineered bacteria capable of producing lysine diol further comprises one or more of the following gene modifications: (1) At least one gene modification that enhances the function of acetyl-CoA transferase; (2) At least one gene modification that enhances the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase; (4) At least one gene modification that enhances the activity of mevalonate kinase; (5) At least one gene modification that enhances the activity of mevalonate-5-phosphate kinase; (6) At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase; (7) At least one gene modification that enhances the activity of pentylene pyrophosphate isomerase; and (8) At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase.

20. The method of claim 19, wherein, At least one gene modification that enhances the function of acetyl-CoA transferase includes: introducing an exogenous acetyl-CoA transferase gene, increasing the gene copy number of the acetyl-CoA transferase gene, and / or replacing the natural promoter of endogenous acetyl-CoA transferase with a promoter that has a higher expression level. At least one gene modification that enhances the function of 3-methyl-3-hydroxyglutaryl-CoA synthase includes: introducing an exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene, increasing the gene copy number of the 3-methyl-3-hydroxyglutaryl-CoA synthase gene, and / or replacing the natural promoter of endogenous 3-methyl-3-hydroxyglutaryl-CoA synthase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase includes: introducing an exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene, increasing the gene copy number of the 3-hydroxy-3-methylglutaryl-CoA reductase gene, and / or replacing the natural promoter of endogenous 3-hydroxy-3-methylglutaryl-CoA reductase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of mevalonate kinase includes: introducing an exogenous mevalonate kinase gene, increasing the gene copy number of the mevalonate kinase gene, and / or replacing the natural promoter of endogenous mevalonate kinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-phosphokinase includes: introducing an exogenous mevalonate-5-phosphokinase gene, increasing the gene copy number of the mevalonate-5-phosphokinase gene, and / or replacing the natural promoter of endogenous mevalonate-5-phosphokinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase includes: introducing an exogenous mevalonate-5-pyrophosphate decarboxylase gene, increasing the gene copy number of the mevalonate-5-pyrophosphate decarboxylase gene, and / or replacing the natural promoter of endogenous mevalonate-5-pyrophosphate decarboxylase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of pentyrene pyrophosphate isomerase includes: introducing an exogenous pentyrene pyrophosphate isomerase gene, increasing the copy number of the pentyrene pyrophosphate isomerase gene, and / or replacing the natural promoter of the endogenous pentyrene pyrophosphate isomerase with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase includes: introducing an exogenous farnesyl pyrophosphate synthase gene, increasing the gene copy number of the farnesyl pyrophosphate synthase gene, and / or replacing the natural promoter of endogenous farnesyl pyrophosphate synthase with a promoter that has a higher expression level.

21. The method of claim 20, wherein: The exogenous acetyl-CoA transferase comprises the amino acid sequence shown in SEQ ID NO.3, preferably, the exogenous acetyl-CoA transferase is encoded by the nucleotide sequence of SEQ ID NO.229; The exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase comprises the amino acid sequence shown in SEQ ID NO.4, preferably, the exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase is encoded by the nucleotide sequence of SEQ ID NO.230; The exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a truncated 3-hydroxy-3-methylglutaryl-CoA reductase, wherein the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is a truncated 3-hydroxy-3-methylglutaryl-CoA reductase with 528 amino acids removed from its N-terminus; preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase comprises the amino acid sequence shown in SEQ ID NO. 2, and preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is encoded by the nucleotide sequence of SEQ ID NO. 228; The exogenous mevalonate kinase comprises the amino acid sequence shown in SEQ ID NO.5, preferably, the exogenous mevalonate kinase is encoded by the nucleotide sequence of SEQ ID NO.231; The exogenous mevalonate-5-phosphokinase comprises the amino acid sequence shown in SEQ ID NO. 6, preferably, the exogenous mevalonate-5-phosphokinase is encoded by the nucleotide sequence of SEQ ID NO. 232; The exogenous mevalonate-5-pyrophosphate decarboxylase comprises the amino acid sequence shown in SEQ ID NO.7, preferably, the exogenous mevalonate-5-pyrophosphate decarboxylase is encoded by the nucleotide sequence of SEQ ID NO.233; The exogenous pentene pyrophosphate isomerase comprises the amino acid sequence shown in SEQ ID NO. 8, preferably, the exogenous pentene pyrophosphate isomerase is encoded by the nucleotide sequence of SEQ ID NO. 234; and / or The exogenous farnesyl pyrophosphate synthase comprises the amino acid sequence shown in SEQ ID NO.1, preferably encoded by the nucleotide sequence of SEQ ID NO.

227.

22. The method according to any one of claims 9-21, wherein the recombinant engineered bacteria capable of producing lysine diol is a bacterium or yeast; preferably, the bacteria are selected from Escherichia, Corynebacterium, or Bacillus, and the yeast is selected from Saccharomyces, Pichia, Hansenula, Kluyveromyces, Phaffia, Schizosaccharomyces, Candida, Yarrowia, Hyphozyma, or Cryptococcus; more preferably, the bacteria are selected from Escherichia coli (…). Escherichia coli ), Corynebacterium glutamicum ( Corynebacterium glutamicum ), or Bacillus subtilis ( Bacillus subtilis The yeast is selected from brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae Pichia pastoris () Pichia pastoris ), Phaffia colomata ( Komagataella phaffii) , Saccharomyces cerevisiae ( Schizosaccharomycespombe ), Candida albicans ( Candidaalbicans ), Candida utilis ( Candida utilis ), Yarrowia lipolytica ( Yarrowia lipolytica ), Hansenula polymorpha ( Hansenula polymorpha Pichia pastoris (Canada) Pichia canadensis ), Max Kluyveromycin ( Kluyveromyces marxianus Kluyveromycin (lactic acid yeast) Kluyveromyces lactis ), Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus albidus ) or Red Pavlova yeast ( Phaffia rhodozyma ).

23. The method according to any one of claims 1-22, comprising co-culturing the microorganism capable of producing lysinediol and the microorganism capable of converting lysinediol into perillyl and / or perillyl lactone under suitable conditions.

24. The method according to any one of claims 1-23, wherein, The cell density ratio of the microorganisms capable of producing lysenoside diol and the microorganisms capable of converting lysenoside diol into perillaldehyde and / or perillaldehyde lactone is in the range of 20:1 to 1:20; preferably, the cell density ratio of the microorganisms capable of producing lysenoside diol and the microorganisms capable of converting lysenoside diol into perillaldehyde and / or perillaldehyde lactone is 10.5:1, 8.5:1, or 4.

8. :1, 3.9:1, 2:1, 1.9:1, 1.55:1, 1:1, 0.96:1, 0.78:1, 1:2, 1:2.1, 1:2.58, 1:5.2, 1:6.45, 1:11.5, 1:14; More preferably, the microorganisms capable of producing lysoprenediol and the microorganisms capable of converting lysoprenediol into perillylene diol and / or perillylene lactone have the accession number ATCC. The cell density ratios of the fungi in the mixture of 20624 are 8.5:1, 3.9:1, 2:1, 1.55:1, 1:1, 0.78:1, 1:2, 1:2.58, and 1:6.45; or the cell density ratios of the fungi in the mixture of the microorganisms capable of producing lysine-1-diol and the microorganisms capable of converting lysine-1-diol into perillyl alcohol and / or perillyl lactone are 10.5:1, 4.8:1, 2:1, 1.9:1, 1:1, 0.96:1, 1:2, 1:2.1, 1:5.2, and 1:11.

5.

25. The method of claim 23 or 24, wherein, The temperature for the mixed culture is above 15°C; preferably, the temperature for the mixed culture is 15°C-40°C, and more preferably, the temperature for the mixed culture is 25°C-30°C.

26. The method according to any one of claims 23-25, wherein the method comprises adding at least one carbon source during the mixed culture process, said at least one carbon source being selected from glucose, fructose, sucrose, acetic acid, glycerol, maltose, lactic acid, and succinic acid.

27. The method according to any one of claims 23-26, wherein, The microorganisms capable of converting lysinediol into perillyl and / or perillyl lactone are either lysinediol-induced or not lysinediol-induced before being mixed with the microorganisms capable of producing lysinediol.

28. The method according to any one of claims 1-27, the method further comprising separating perillaldehyde and / or perillyl lactone.

29. A composition comprising two microorganisms: First microorganism: Selected from microorganisms capable of producing lysine diol; Second microorganism: Selected from microorganisms capable of converting lysine-enrichediol into perillyl alcohol and / or perillyl lactone.

30. The composition of claim 29, wherein, The microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from the genera *Hyphozyma* or *Cryptococcus*; preferably, the microorganisms capable of converting lysine-enriched diol into perillyl diol and / or perillyl lactone are selected from the genera *Hyphozyma* or *Cryptococcus*. Hyphozyma roseoniger Or Cryptococcus lightensis ( Cryptococcus albidus ).

31. The composition of claim 30, wherein The microorganisms capable of converting lysine-1-diol into perillyl alcohol and / or perillyl lactone are selected from fungi with accession number ATCC 20624 or fungi with accession number ATCC 20918 or their derivatives.

32. The composition according to any one of claims 29-31, wherein, The first microorganism contains a phosphorylase that catalyzes the formation of lysandrindiol pyrophosphate (LPP) to lysandrindiol (LOH). This phosphorylase is derived from Mg(2+)-dependent phosphatidate phosphatase, phosphatidic acid phosphatase type 2, serine / threonine-protein phosphatase, HAD-like hydrolase superfamily, phosphate (PA) phosphatase, polyphosphoinositide phosphatase, serine / threonine-protein kinases, alkaline phosphatase, or phosphoprotein phosphatase family. This includes various protein phosphatases, serine / threonine-protein phosphatases, the SIT4 phosphatase-associated protein family, the TAP42 / TAP46-like superfamily, dual-specificity lipid and protein phosphatases, polynucleotide kinase 3 phosphatases, 5'-deoxynucleotidases, myosin family, histidine phosphatase superfamily, S-2-haloalkanoic acid dehalogenase, phosphatidylglycerol phosphatase, and haloic acid dehalogenase (HAD) hydrolase superfamily. ), phosphatase family (PA-phosphatase)Related phosphoesterase family, squalene / phytoene synthase family, terpene cyclase family, phosphatase App1, protein phosphatase 2A (PP2A), protein phosphatase 2C (PP2C), halogen dehalogenase (HAD), sugar phosphophosphate phosphatase (HAD-like), polyphosphoinositide phosphatase (Fig4-like), casein kinase 1 (Se / Thr protein kinase), lipoprotein family, PPZ / Ppq1 family, clade-1 family, HAD-IA family hydrolases (HAD-IA family). The enzymes are preferably phosphatases such as IA hydrolase family, HAD-IF subfamily (IF family), dolichoyldiphosphatase family, or haloperoxidase superfamily; preferably, the phosphatase is phosphatidylic acid phosphatase (APP1), dolichoyldiphosphatase (CAX4), PP2A protein phosphatase regulatory subunit B (CDC55), 2-deoxyglucose-6-phosphatase 1 (DOG1), diacylglycerol pyrophosphate phosphatase 1 (DPP1), inositol polyphosphate phosphatase (FIG4), 1-glycerol phosphate phosphatase 1 (GPP1), casein kinase I (HRR25), phosphatidylic acid phosphatase (PAH1), alkaline phosphatase (PHO8), truncated alkaline phosphatase (PHO8^62aa), serine / threonine protein phosphatase (PPQ), serine / threonine protein phosphatase (PPZ1), or serine / threonine protein phosphatase (PTC3). Serine / threonine protein phosphatase (PTC3), SIT4-associated protein SAP155, SIT4-associated protein SAP185, type 2A phosphatase-associated protein (TAP42), phosphatidylinositol 3,4,5-trisphosphate phosphatase (TEP1), polynucleotides3'-phosphatase (TPP1), phosphorylhydrolase (YAR068W), 5'-deoxynucleotidase (YBR242W), phosphatidylinositol 3-phosphatase, phosphorylhydrolase (YNL108C), hydrolase (YOR131C), histidine phosphatase family phosphatase (YOR283W), phosphatidylglycerol phosphatase (PgpA), phosphatidylglycerol phosphatase (PgpB), phosphatidylglycerol phosphatase (PgpC), carbapenyl diphosphatase (YbjG), dihydroxy diphosphatase (DOLPP1), phosphorylhydrolase (PLPP6), farnesyl diphosphatase (YisP), farnesol synthase (TPS2), acyclic sesquiterpene synthase (TPS1), or farnesol synthase (TPS13).

33. The composition according to any one of claims 29-32, wherein, The phosphate hydrolase is a phosphate hydrolase derived from microorganisms or its functional variants, including bacteria, yeast, or fungi, wherein the bacteria are selected from Escherichia or Bacillus, the yeast is selected from Saccharomyces, or the phosphate hydrolase is derived from plants or humans. Homo sapiens Phosphohydrolases or their functional variants thereof; preferably, the phosphohydrolase is a phosphohydrolase or its functional variant derived from bacteria, yeast or fungi, wherein the bacteria are selected from *Escherichia coli* (…). Escherichia coli Bacillus subtilis ( Bacillus subtilis The yeast is brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), or the phosphorylase is derived from plants or humans ( Homo sapiens Phosphorylase or a functional variant thereof, wherein the plant is selected from moso bamboo ( Phyllostachys edulis ),corn( Zea mays ) or japonica rice ( Oryza sativa subsp. japonica More preferably, the phosphorylase is derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae). Saccharomyces cerevisiae ), Escherichia coli ( Escherichia coli ), human beings ( Homo sapiens Bacillus subtilis ( Bacillus subtilis ),bamboo( Phyllostachys edulis ),corn( Zea mays ) or japonica rice ( Oryza sativa subsp. japonica Acid hydrolases or their functional variants.

34. The composition according to any one of claims 29-33, wherein the phosphorylase comprises an amino acid sequence as shown in SEQ ID NO. 11-226, or an amino acid sequence that is at least 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% identical to the amino acid sequence shown in SEQ ID NO. 12, SEQ ID NO. 17, SEQ ID NO. 20, SEQ ID NO. 36, SEQ ID NO. 38, SEQ ID NO. 45, SEQ ID NO. 58, SEQ ID NO. 63 ...65, SEQ ID NO. 66, SEQ ID NO. 67, SEQ ID NO. 68, SEQ ID NO. 69, SEQ ID NO. 69, SEQ ID NO. 60, SEQ ID NO. 69, SEQ ID NO. 60, ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ ID NO.159, SEQ ID NO.160, SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID The amino acid sequence of NO.225, SEQ ID NO.226, or selected from SEQ ID NO.12, SEQ ID NO.17, SEQ ID NO.20, SEQ ID NO.36, SEQ ID NO.38, SEQ ID NO.45, SEQ ID NO.58, SEQ ID NO.63, SEQ ID NO.95, SEQ ID NO.106, SEQ ID NO.107, SEQ ID NO.122, SEQ ID NO.125, SEQ ID NO.133, SEQ ID NO.157, SEQ IDNO.159, SEQ ID NO.

160. SEQ ID NO.180, SEQ ID NO.181, SEQ ID NO.186, SEQ ID NO.197, SEQ ID NO.198, SEQ ID NO.206, SEQ ID NO.208, SEQ ID NO.209, SEQ ID NO.210, SEQ ID NO.217, SEQ ID NO.218, SEQ ID NO.219, SEQ ID NO.220, SEQ ID NO.221, SEQ ID NO.222, SEQ ID NO.223, SEQ ID NO.224, SEQ ID NO.225, SEQ ID The amino acid sequence of NO. 226 has at least 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% sequence identity.

35. The composition according to any one of claims 29-34, wherein the phosphorylase is encoded by a nucleotide sequence selected from those shown in SEQ ID NO. 237-452, or by a nucleotide sequence selected from those with at least 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% sequence identity with the nucleotide sequence shown in SEQ ID NO. 237-452; preferably, the phosphorylase is encoded by a nucleotide sequence selected from SEQ ID NO. 238, SEQ ID NO. 243, SEQ ID NO. 246, SEQ ID NO. 262, SEQ ID NO. 264, SEQ ID NO. 271, SEQ ID NO. 242 ...4, SEQ ID NO. 245, SEQ ID NO. 243, SEQ ID NO. 246, SEQ ID NO. 242, NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.383, SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID The nucleotide sequence of NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ ID NO.452, or selected from the group consisting of SEQ ID NO.238, SEQ ID NO.243, SEQ ID NO.246, SEQ ID NO.262, SEQ ID NO.264, SEQ ID NO.271, SEQ ID NO.284, SEQ ID NO.289, SEQ ID NO.321, SEQ ID NO.332, SEQ ID NO.333, SEQ ID NO.348, SEQ ID NO.351, SEQ ID NO.359, SEQ ID NO.

383. SEQ ID NO.385, SEQ ID NO.386, SEQ ID NO.406, SEQ ID NO.407, SEQ ID NO.412, SEQ ID NO.423, SEQ ID NO.424, SEQ ID NO.432, SEQ ID NO.434, SEQ ID NO.435, SEQ ID NO.436, SEQ ID NO.443, SEQ ID NO.444, SEQ ID NO.445, SEQ ID NO.446, SEQ ID NO.447, SEQ ID NO.448, SEQ ID NO.449, SEQ ID NO.450, SEQ ID NO.451, SEQ ID The nucleotide sequence of NO. 452 encodes a nucleotide sequence with at least 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% sequence identity.

36. The composition according to any one of claims 29-35, wherein the first microorganism comprises a gene for the lysine pyrophosphate (LPP) synthesis pathway.

37. The composition of claim 36, wherein the lysine pyrophosphate (LPP) synthesis pathway gene comprises a farnesyl pyrophosphate (FPP) synthesis pathway gene, a geraniol geraniol pyrophosphate synthase (GGPPS) gene, and / or a lysine pyrophosphate synthase (LPPS) gene.

38. The composition of claim 37, wherein the farnesyl pyrophosphate (FPP) synthesis pathway gene comprises one or more of the following: acetyl-CoA transferase gene, 3-methyl-3-hydroxyglutaryl-CoA synthase gene, 3-hydroxy-3-methylglutaryl-CoA reductase gene, mevalonate kinase gene, mevalonate-5-phosphate kinase gene, mevalonate-5-pyrophosphate decarboxylase gene, pentene pyrophosphate isomerase gene, or farnesyl pyrophosphate synthase gene.

39. The composition according to any one of claims 29-38, wherein the first microorganism further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the activity of geraniol geraniol pyrophosphate synthase (GGPPS); and (2) At least one gene modification that enhances the activity of lysine pyrophosphate diol ester synthase (LPPS).

40. The composition of claim 39, wherein, At least one gene modification that enhances the activity of gerany-gerany-gerany pyrophosphate synthase (GGPPS) includes: introducing an exogenous gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, increasing the gene copy number of the gerany-gerany-gerany pyrophosphate synthase (GGPPS) gene, and / or replacing the natural promoter of the endogenous gerany-gerany pyrophosphate synthase (GGPPS) with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of lysandrindiol pyrophosphate synthase (LPPS) includes: introducing an exogenous lysandrindiol pyrophosphate synthase (LPPS) gene, increasing the gene copy number of the lysandrindiol pyrophosphate synthase (LPPS) gene, and / or replacing the natural promoter of the endogenous lysandrindiol pyrophosphate synthase (LPPS) with a promoter that has a higher expression level.

41. The composition of claim 40, wherein, The exogenous gerany-gerany pyrophosphate synthase (GGPPS) comprises the amino acid sequence shown in SEQ ID NO. 9, preferably encoded by the nucleotide sequence of SEQ ID NO. 235; and / or The exogenous lysine pyrophosphate diol ester synthase (LPPS) gene encodes a truncated lysine pyrophosphate diol ester synthase (LPPS); preferably, the truncated lysine pyrophosphate diol ester synthase (LPPS) is a signal peptide with the first 63 amino acids removed; preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) comprises the amino acid sequence shown in SEQ ID NO. 10, more preferably, the exogenous lysine pyrophosphate diol ester synthase (LPPS) is encoded by the nucleotide sequence of SEQ ID NO.

236.

42. The composition according to any one of claims 29-41, wherein the first microorganism further comprises one or more of the following genetic modifications: (1) At least one gene modification that enhances the function of acetyl-CoA transferase; (2) At least one gene modification that enhances the activity of 3-methyl-3-hydroxyglutaryl-CoA synthase; (3) At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase; (4) At least one gene modification that enhances the activity of mevalonate kinase; (5) At least one gene modification that enhances the activity of mevalonate-5-phosphate kinase; (6) At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase; (7) At least one gene modification that enhances the activity of pentylene pyrophosphate isomerase; and (8) At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase.

43. The composition of claim 42, wherein, At least one gene modification that enhances the function of acetyl-CoA transferase includes: introducing an exogenous acetyl-CoA transferase gene, increasing the gene copy number of the acetyl-CoA transferase gene, and / or replacing the natural promoter of endogenous acetyl-CoA transferase with a promoter that has a higher expression level. At least one gene modification that enhances the function of 3-methyl-3-hydroxyglutaryl-CoA synthase includes: introducing an exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase gene, increasing the gene copy number of the 3-methyl-3-hydroxyglutaryl-CoA synthase gene, and / or replacing the natural promoter of endogenous 3-methyl-3-hydroxyglutaryl-CoA synthase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of 3-hydroxy-3-methylglutaryl-CoA reductase includes: introducing an exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene, increasing the gene copy number of the 3-hydroxy-3-methylglutaryl-CoA reductase gene, and / or replacing the natural promoter of endogenous 3-hydroxy-3-methylglutaryl-CoA reductase with a promoter that has a higher expression level; At least one gene modification that enhances the activity of mevalonate kinase includes: introducing an exogenous mevalonate kinase gene, increasing the gene copy number of the mevalonate kinase gene, and / or replacing the natural promoter of endogenous mevalonate kinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-phosphokinase includes: introducing an exogenous mevalonate-5-phosphokinase gene, increasing the gene copy number of the mevalonate-5-phosphokinase gene, and / or replacing the natural promoter of endogenous mevalonate-5-phosphokinase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of mevalonate-5-pyrophosphate decarboxylase includes: introducing an exogenous mevalonate-5-pyrophosphate decarboxylase gene, increasing the gene copy number of the mevalonate-5-pyrophosphate decarboxylase gene, and / or replacing the natural promoter of endogenous mevalonate-5-pyrophosphate decarboxylase with a promoter that has a higher expression level. At least one gene modification that enhances the activity of pentyrene pyrophosphate isomerase includes: introducing an exogenous pentyrene pyrophosphate isomerase gene, increasing the copy number of the pentyrene pyrophosphate isomerase gene, and / or replacing the natural promoter of the endogenous pentyrene pyrophosphate isomerase with a promoter that has a higher expression level; and / or At least one gene modification that enhances the activity of farnesyl pyrophosphate synthase includes: introducing an exogenous farnesyl pyrophosphate synthase gene, increasing the gene copy number of the farnesyl pyrophosphate synthase gene, and / or replacing the natural promoter of endogenous farnesyl pyrophosphate synthase with a promoter that has a higher expression level.

44. The composition of claim 43, wherein: The exogenous acetyl-CoA transferase comprises the amino acid sequence shown in SEQ ID NO.3, preferably, the exogenous acetyl-CoA transferase is encoded by the nucleotide sequence of SEQ ID NO.229; The exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase comprises the amino acid sequence shown in SEQ ID NO.4, preferably, the exogenous 3-methyl-3-hydroxyglutaryl-CoA synthase is encoded by the nucleotide sequence of SEQ ID NO.230; The exogenous 3-hydroxy-3-methylglutaryl-CoA reductase gene encodes a truncated 3-hydroxy-3-methylglutaryl-CoA reductase, wherein the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is a truncated 3-hydroxy-3-methylglutaryl-CoA reductase with 528 amino acids removed from its N-terminus; preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase comprises the amino acid sequence shown in SEQ ID NO. 2, and preferably, the truncated 3-hydroxy-3-methylglutaryl-CoA reductase is encoded by the nucleotide sequence of SEQ ID NO. 228; The exogenous mevalonate kinase comprises the amino acid sequence shown in SEQ ID NO.5, preferably, the exogenous mevalonate kinase is encoded by the nucleotide sequence of SEQ ID NO.231; The exogenous mevalonate-5-phosphokinase comprises the amino acid sequence shown in SEQ ID NO. 6, preferably, the exogenous mevalonate-5-phosphokinase is encoded by the nucleotide sequence of SEQ ID NO. 232; The exogenous mevalonate-5-pyrophosphate decarboxylase comprises the amino acid sequence shown in SEQ ID NO.7, preferably, the exogenous mevalonate-5-pyrophosphate decarboxylase is encoded by the nucleotide sequence of SEQ ID NO.233; The exogenous pentene pyrophosphate isomerase comprises the amino acid sequence shown in SEQ ID NO. 8, preferably, the exogenous pentene pyrophosphate isomerase is encoded by the nucleotide sequence of SEQ ID NO. 234; and / or The exogenous farnesyl pyrophosphate synthase comprises the amino acid sequence shown in SEQ ID NO.1, preferably encoded by the nucleotide sequence of SEQ ID NO.

227.

45. The composition according to any one of claims 29-44, wherein the first microorganism is a bacterium or yeast; preferably, the bacterium is selected from Escherichia, Corynebacterium, or Bacillus, and the yeast is selected from Saccharomyces, Pichia, Hansenula, Kluyveromyces, Phaffia, Schizosaccharomyces, Candida, Yarrowia, Hyphozyma, or Cryptococcus; more preferably, the bacterium is selected from Escherichia coli (…). Escherichia coli ), Corynebacterium glutamicum ( Corynebacterium glutamicum ), or Bacillus subtilis ( Bacillus subtilis The yeast is selected from brewer's yeast (Saccharomyces cerevisiae). Saccharomyces cerevisiae Pichia pastoris () Pichia pastoris ), Phaffia colomata ( Komagataellaphaffii) , Saccharomyces cerevisiae ( Schizosaccharomycespombe ), Candida albicans ( Candidaalbicans ), Candida utilis ( Candida utilis ), Yarrowia lipolytica ( Yarrowia lipolytica ), Hansenula polymorpha ( Hansenulapolymorpha Pichia pastoris (Canada) Pichia canadensis Kluyveromyces marxianus and Kluyveromyces lactis. Hyphozyma roseoniger Cryptococcus syriacus ( Cryptococcus albidus ) or Red Pavlova yeast ( Phaffia rhodozyma ).

46. ​​Use of the composition according to any one of claims 29-45 in the production of perillaldehyde and / or perillaldehyde lactone.

47. Use of the fungus or its derivative with accession number ATCC 20918 in the production of perillaldehyde and / or perillol from lysine-1-diol.

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