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Phytases, nucleic acids encoding them and methods for making and using them
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A phytase and nucleic acid technology, which can be used in hydrolase, medical preparations containing active ingredients, bioreactors/fermenters for specific purposes, etc., and can solve problems such as insufficient optimization and waste.
Inactive Publication Date: 2012-09-05
BASF ENZYMES
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[0007] 2) The presence of unhydrolyzed phytate in many ex vivo applications is undesirable and problematic (e.g. leading to the appearance of unwanted precipitates)
[0010] 5) Nutritious ingredients provided in many potential foods are not fully utilized and are wasted
[0028] To solve the mentioned problems, it has been proposed to treat food with exogenous phytase, but this method has not been fully optimized, especially with regard to feasibility and cost-effectiveness
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Embodiment 1
[0183] Alternatively, these nucleic acids can be synthesized in vitro by well-known chemical synthesis techniques, as described in, e.g., Adams (1983) J. Am. Chem. Soc. 105:661; Belousov (1997) Nucleic Acids Res. 25: 3440-3444; Frenkel (1995) Free Radic. Biol. Med. 19: 373-380; Blommers (1994) Biochemistry 33: 7886-7896; Narang (1979) Meth. Enzymol. 68: 90; Brown (1979) Meth. Enzymol. 68:109; Beaucage (1981) Tetra. Lett. 22:1859; US Patent No. 4,458,066.
Embodiment 2
[0184] Techniques for manipulating nucleic acids such as subcloning, labeling probes (e.g., labeling random primers using Klenow polymerase, nick translation, amplification), sequencing, hybridization, and similar techniques are well described in the academic and patent literature, see, For example, edited by Sambrook, Molecular Cloning (MOLECULAR CLONING: a LABORATORY MANUAL): Laboratory Manual (Second Edition), Vols.1-3, Cold Spring Harbor Laboratory, (1989); Modern Methods of Molecular Biology, (CURRENT PROTOCOLS IN MOLECULAR BIOLOGY) Ausubel ed., John Wiley & Sons, Inc., New York (1997); Biochemistry and Molecular Biology Laboratory Technology (LABORATORY TECHNIQUES INBIOCHEMISTRY AND MOLECULAR BIOLOGY: HYBRIDIZATION WITH NUCLEIC ACID PROBES): Hybridization with Nucleic Acid Probes, pp. Part. Theory and Nucleic Acid Preparation, edited by Tijssen, Elsevier, N.Y. (1993).
Embodiment 3
[0184] Techniques for manipulating nucleic acids such as subcloning, labeling probes (e.g., labeling random primers using Klenow polymerase, nick translation, amplification), sequencing, hybridization, and similar techniques are well described in the academic and patent literature, see, For example, edited by Sambrook, Molecular Cloning (MOLECULAR CLONING: a LABORATORY MANUAL): Laboratory Manual (Second Edition), Vols.1-3, Cold Spring Harbor Laboratory, (1989); Modern Methods of Molecular Biology, (CURRENT PROTOCOLS IN MOLECULAR BIOLOGY) Ausubel ed., John Wiley & Sons, Inc., New York (1997); Biochemistry and Molecular Biology Laboratory Technology (LABORATORY TECHNIQUES INBIOCHEMISTRY AND MOLECULAR BIOLOGY: HYBRIDIZATION WITH NUCLEIC ACID PROBES): Hybridization with Nucleic Acid Probes, pp. Part. Theory and Nucleic Acid Preparation, edited by Tijssen, Elsevier, N.Y. (1993).
[0185] Another useful method of obtaining and manipulating nucleic acids for carrying out the methods ...
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Abstract
The invention provides isolated and recombinant phytase enzymes. In one aspect, the phytases are produced by modification of the wild type appA of E. coli. The enzyme can be produced from recombinant host cells. The phytases of the invention can be used to aid in the digestion of phytate where desired. In particular, the phytases of the invention can be used in foodstuffs to improve the feeding value of phytate rich ingredients. The phytases of the invention can be thermotolerant and / or thermostable. Also provided are methods for obtaining a variant polynucleotide encoding a phytase and for obtaining a phytase with thermostability or thermotolerant at high or low temperatures.
Description
[0001] Cross References to Related Applications [0002] This application is a continuation of US Patent Application No. (USSN) 09 / 866,379, filed May 24, 2001. technical field [0003] The present invention relates to newly prepared polynucleotides, polypeptides encoded by such polynucleotides, uses of such polynucleotides and polypeptides, and production and isolation of such polynucleotides and polypeptides. In particular, the present invention provides a polypeptide having phytase activity, such as SEQ ID NO:1. The present invention provides isolated and recombinant phytases. Phytases can be produced by modifying E. coli wild-type appA or from recombinant host cells. The phytases of the invention can be used to aid in the digestion of phytic acid. The phytases of the invention can be used in food products to improve the feed value of phytate-rich ingredients. The phytases of the invention may be thermotolerant and / or thermostable. Also provided are methods of obtainin...
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