Methods and compositions for producing fatty aldehydes
View Patent ↗Methods and compositions, including nucleotide sequences, amino acid sequences, and host cells, for producing fatty aldehydes are described.
1. A method of producing a fatty aldehyde, the method comprising:
(a) providing a bacterial host cell, said bacterial host cell engineered to express a gene encoding a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 22 (carB), and comprising the phosphopantetheine attachment site sequence of SEQ ID NO: 9, wherein the polypeptide has carboxylic acid reductase activity effective to convert a fatty acid to a fatty aldehyde; and
(b) culturing said engineered bacterial host cell in a culture media containing a carbohydrate carbon source under conditions effective to produce a fatty aldehyde at a yield of at least 25 mg/L in the media of the bacterial host cell culture, when cultured for 20 hours at 37° C.
2. The method of claim 1 , further comprising modifying the expression of a gene encoding a polypeptide having fatty acid synthase activity in the bacterial host cell.
3. The method of claim 2 , wherein said gene encodes a polypeptide having thioesterase activity.
4. The method of claim 1 , wherein the bacterial host cell is genetically engineered to express an attenuated level of a fatty acid degradation enzyme relative to a wild type bacterial host cell.
5. A method of producing a fatty aldehyde, the method comprising:
(a) providing a bacterial host cell, engineering said bacterial host cell engineered to comprise a recombinant vector comprising a nucleotide sequence having the nucleotide sequence of SEQ ID NO: 21, wherein the nucleotide sequence encodes a polypeptide having carboxylic acid reductase activity which comprises the phosphopantetheine attachment site sequence of SEQ ID NO: 9, and
(b) culturing said engineered bacterial host cell in a culture media containing a carbohydrate carbon source under conditions effective to produce a fatty aldehyde at a yield of at least 25 mg/L in the media of the bacterial host cell culture, when cultured for 20 hours at 37° C.
6. The method of claim 5 , further comprising modifying the expression of a gene encoding a polypeptide having fatty acid synthase activity in the bacterial host cell.
7. The method of claim 6 , wherein said gene encodes a polypeptide having thioesterase activity.
8. The method of claim 5 , wherein the bacterial host cell is a genetically engineered bacterial host cell that expresses an attenuated level of a fatty acid degradation enzyme relative to a wild type bacterial host cell, wherein the fatty acid degradation enzyme has acyl-CoA synthase (EC 2.3.1.86) activity.
9. The method of claim 1 , wherein the fatty aldehyde is isolated from the extracellular environment of the bacterial host cell.
10. The method of claim 1 , wherein the fatty aldehyde comprises a C 6 -C 26 fatty aldehyde.
11. The method of claim 10 , wherein the fatty aldehyde is decanal, dodecanal, myristal, or hexadecal.
12. The method of claim 1 , wherein the fatty aldehyde is an unsaturated fatty aldehyde.
13. The method of claim 1 , wherein the fatty aldehyde is a saturated fatty aldehyde.
14. The method of claim 1 , wherein said carbohydrate carbon source is a monosaccharide.
15. The method of claim 14 , wherein said monosaccharide is glucose.
16. The method of claim 15 , wherein the fatty aldehyde is extracted from the culture media.
17. The method of claim 1 , wherein the polypeptide comprises the reductase domain sequence of SEQ ID NO: 7, the NADP binding domain sequence of SEQ ID NO: 8 and the AMP binding domain sequence of SEQ ID NO: 10.
18. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 22.
19. The method of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 22.
20. The method of claim 4 , wherein the fatty acid degradation enzyme has acyl-CoA synthase (EC 2.3.1.86) activity.