Method for producing oxo fatty acid and rare fatty acid
The present invention provides a production method of oxo fatty acid, as well as rare fatty acids such as conjugated fatty acid, hydroxylated fatty acid, partially saturated fatty acid and the like, which uses 4 kinds of enzymes (fatty acid-hydratase, hydroxylated fatty acid-dehydrogenase, oxo fatty acid-isomerase, oxo fatty acid-enone reductase) derived from Lactobacillus plantarum including lactic acid bacteria and the like. Furthermore, the present invention also provides a more efficient production method of oxo fatty acid and the like, which partly uses a chemical oxidation reaction in combination.
1. A method of producing an oxo fatty acid having 18 carbon atoms and a carbonyl group at the 10-position, and not having a double bond at the 11- and 12-positions comprising contacting an oxo fatty acid having 18 carbon atoms, a carbonyl group at the 10-position and a trans-type double bond at the 11-position and a saturase to form an oxo fatty acid having 18 carbon atoms and a carbonyl group at the 10-position, and not having a double bond at the 11- and 12-positions, wherein the saturase is
(i) obtained from Lactobacillus plantarum FERM BP-10549 strain, or
(ii) an enzyme protein of any of the following (a) - (c):
(a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2,
(b) a protein comprising the amino acid sequence shown in SEQ ID NO: 2, wherein 1-20 amino acids are deleted, substituted, inserted, and/or added, and having an enzyme activity of catalyzing the saturation reaction, or
(c) a protein encoded by a nucleotide sequence that hybridizes to the complement of the nucleotide sequence shown in SEQ ID NO: 1 under conditions of washing 2-3 times with a buffer comprising 0.1×SSC and 0.1% SDS at 68° C., and having an enzyme activity to catalyze the saturation reaction.
2. The method according to claim 1 , wherein the oxo fatty acid having 18 carbon atoms, a carbonyl group at the 10-position and a trans-type double bond at the 11-position is 10-oxo-trans-11-octadecenoic acid, 10-oxo-cis-6,trans-11, -octadecadienoic acid, 10-oxo-trans-11,cis-15-octadecadienoic acid or 10-oxo-ci s-6,trans -11,cis-15-octadecatrienoic acid.
3. The method according to claim 1 , wherein the saturase is obtained from Lactobacillus plantarum FERM BP-10549 strain.
4. The method according to claim 1 , wherein the saturase is an enzyme protein of any of the following (a) - (c):
(a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2,
(b) a protein comprising the amino acid sequence shown in SEQ ID NO: 2, wherein 1-20 amino acids are deleted, substituted, inserted, and/or added, and having an enzyme activity of catalyzing the saturation reaction, or
(c) a protein encoded by a nucleotide sequence that hybridizes to the complement of the nucleotide sequence shown in SEQ ID NO: 1 under conditions of washing 2-3 times with a buffer comprising 0.1×SSC and 0.1% SDS at 68° C., and having an enzyme activity to catalyze the saturation reaction in claim 1 .
5. The method according to claim 1 , wherein the saturase is an enzyme protein consisting of the amino acid sequence shown in SEQ ID NO: 2.
6. The method according to claim 1 , wherein the saturase is a protein comprising the amino acid sequence shown in SEQ ID NO: 2, wherein 1-20 amino acids are deleted, substituted, inserted, and/or added, and having an enzyme activity of catalyzing the saturation reaction.
7. The method according to claim 1 , wherein the saturase is a protein comprising an amino acid sequence shown in SEQ ID NO: 2, wherein 1-10 amino acids are deleted, substituted, inserted, and/or added, and having an enzyme activity of catalyzing the saturation reaction.
8. The method according to claim 4 , wherein the saturase is a protein encoded by a nucleotide sequence that hybridizes to the complement of the nucleotide sequence shown in SEQ ID NO: 1 under conditions of washing 2-3 times with a buffer comprising 0.1×SSC and 0.1% SDS at 68° C., and having an enzyme activity to catalyze the saturation reaction.