Method for producing rare fatty acid using novel enzyme, and novel rare fatty acid
The present invention provides production of hydroxylated fatty acid by a hydration reaction using a novel enzyme derived from Lactobacillus and using fatty acid as a substrate, and further, a production method of oxo fatty acid by an enzyme reaction or chemical oxidation reaction using the hydroxylated fatty acid as a substrate. In addition, a valuable novel rare fatty acid obtained by such production method is also provided.
1. 14-Hydroxy-cis-4,cis-7,cis-10,cis-16,cis-19-docosapentaenoic acid.
2. A method of producing 14-hydroxy-cis-4,cis-7,cis-10,cis-16,cis-19-docosapentaenoic acid, comprising incubating under suitable conditions cis-4,cis-7,cis-10,cis-13,cis-16,cis-19-docosahexaenoic acid with a hydratase of any of the following (a)-(c):
(a) the protein consisting of the amino acid sequence shown in SEQ ID NO: 2,
(b) a protein comprising an amino acid sequence wherein one to twenty amino acids in the amino acid sequence shown in SEQ ID NO: 2 are deleted and/or substituted and/or inserted and/or added, and having an enzyme activity that catalyzes a hydration reaction, and
(c) a protein encoded by a nucleotide sequence that hybridizes to a nucleic acid consisting of a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1 under conditions comprising a wash in 0.1×SSC, 0.1% SDS at 68° C., and having an enzyme activity that catalyzes a hydration reaction.
3. A method of producing 14-hydroxy-cis-4,cis-7,cis-10,cis-16,cis-19-docosapentaenoic acid according to claim 2 , wherein the hydratase is the protein consisting of the amino acid sequence shown in SEQ ID NO: 2.
4. A method of producing 14-hydroxy-cis-4,cis-7,cis-10,cis-16,cis-19-docosapentaenoic acid according to claim 2 , wherein the hydratase is a protein comprising an amino acid sequence wherein one to twenty amino acids in the amino acid sequence shown in SEQ ID NO: 2 are deleted and/or substituted and/or inserted and/or added, and having an enzyme activity that catalyzes a hydration reaction.
5. A method of producing 14-hydroxy-cis-4,cis-7,cis-10,cis-16,cis-19-docosapentaenoic acid according to claim 2 , wherein the hydratase is a protein encoded by a nucleotide sequence that hybridizes to a nucleic acid consisting of a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1 under conditions comprising a wash in 0.1×SSC, 0.1% SDS at 68° C., and having an enzyme activity that catalyzes a hydration reaction.