Methods and compositions for producing fatty alcohols
View Patent ↗Methods and compositions, including nucleotide sequences, amino acid sequences, and host cells, for producing fatty alcohols are described.
1. A bacterial cell culture comprising:
(a) a bacterial cell genetically engineered to express an exogenous polypeptide having the amino acid sequence of SEQ ID NO:22, and
(b) a glycerol-containing carbon source,
wherein a fatty alcohol composition comprising C12 and C14 fatty alcohols is produced by the genetically engineered bacterial cell.
2. The cell culture of claim 1 , wherein the bacterial cell is further engineered to express an exogenous polypeptide having thioesterase (EC 3.1.2.14 or EC 3.1.1.5) activity effective to release a fatty acid from acyl-ACP.
3. The cell culture of claim 1 , wherein acyl-CoA dehydrogenase (fadE; EC 1.3.99.3, 1.3.99.) expression or activity is attenuated in the bacterial cell as compared to a wild-type bacterial cell.
4. The cell culture of claim 1 , wherein acyl-CoA dehydrogenase (YdiO; EC 1.3.99.) expression or activity is deleted or reduced in the bacterial cell as compared to a wild-type bacterial cell.
5. The cell culture of claim 1 , wherein 3-ketoacyl-CoA thiolase (fadA; EC 2.3.1.16) expression or activity is deleted or reduced in the bacterial cell as compared to a wild-type bacterial cell.
6. The cell culture of claim 1 , wherein fadB (EC 4.2.1.17, EC 5.1.2.3, EC 5.3.3.8, EC 1.1.1.35) expression or activity is deleted or reduced in the bacterial cell as compared to a wild-type bacterial cell.
7. The cell culture of claim 1 , wherein beta-ketoacyl-CoA thiolase (fadl; EC 2.3.1.16) expression or activity is deleted or reduced in the bacterial cell as compared to a wild-type bacterial cell.
8. The cell culture of claim 1 , wherein fadJ (EC 5.1.2.3, EC 4.2.1.17, EC 1.1.1.35) expression or activity is deleted or reduced in the bacterial cell as compared to a wild-type bacterial cell.
9. A bacterial cell genetically engineered to express
(a) an exogenous polypeptide having the amino acid sequence of SEQ ID NO:22,
(b) a polypeptide having thioesterase (EC 3.1.2.14 or EC 3.1.1.5) activity, and
(c) a polypeptide having attenuated acyl-CoA dehydrogenase (fadE; EC 1.3.99.3, EC 1.3.99.) activity,
wherein the genetically engineered bacterial cell produces a fatty alcohol composition comprising C12 and C14 fatty alcohols when cultured in the presence of a glycerol-containing carbon source.
10. A method of making a fatty alcohol composition, comprising:
a. culturing a bacterial cell genetically engineered to express an exogenous polypeptide having the amino acid sequence of SEQ ID NO:22 in a culture medium comprising glycerol;
b. fermenting the genetically engineered bacterial cell of part a in a fermentation media, wherein a fatty alcohol composition comprising C12 and C14 fatty alcohols is produced by the genetically engineered bacterial cell; and
c. optionally separating the fatty alcohol composition from the fermentation media.
11. The method of claim 10 , wherein the genetically engineered bacterial cell is further engineered to express an exogenous polynucleotide sequence encoding a polypeptide having thioesterase (EC 3.1.2.14 or EC 3.1.1.5) activity effective to release a fatty acid from acyl-ACP.
12. The method of claim 10 , wherein the culture medium further comprises a fatty acid.
13. The method of claim 10 , wherein, acyl-CoA dehydrogenase (fadE; EC 1.3.99.3, 1.3.99.) expression or activity is attenuated in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.
14. The method of claim 10 , wherein acyl-CoA dehydrogenase (Y diO; EC 1.3.99.) expression or activity is deleted or reduced in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.
15. The method of claim 10 , wherein 3-ketoacyl-CoA thiolase (fadA; EC 2.3.1.16) expression or activity is deleted or reduced in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.
16. The method of claim 10 , wherein fadB (EC 4.2.1.17, 5.1.2.3, 5.3.3.8, 1.1.1.35) expression or activity is deleted or reduced in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.
17. The method of claim 10 , wherein beta-ketoacyl-CoA thiolase (fadl; EC 2.3.1.16) expression or activity is deleted or reduced in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.
18. The method of claim 10 , wherein fadJ (EC 5.1.2.3, 4.2.1.17, 1.1.1.35) expression or activity is deleted or reduced in the genetically engineered bacterial cell as compared to a wild-type bacterial cell.