Production of fatty acid derivatives

Patent metadata

Field	Value
Publication number	US-11060099-B2
Application number	US-201715650285-A
Country	US
Kind code	B2
Filing date	Jul 14, 2017
Priority date	Apr 2, 2012
Publication date	Jul 13, 2021
Grant date	Jul 13, 2021

Abstract

The invention relates to compositions and methods, including polynucleotide sequences, amino acid sequences, recombinant host cells and recombinant host cell cultures engineered to produce fatty acid derivative compositions comprising fatty acids, fatty alcohols, fatty aldehydes, fatty esters, alkanes, terminal olefins, internal olefins or ketones. The fatty acid derivative composition is produced extracellularly with a higher titer, yield or productivity than the corresponding wild type or non-engineered host cell.

First claim

We claim:

1. A recombinant host cell comprising a transposon insertion in a yijP gene, wherein the yijP gene is flanked by a gene encoding a phosphoenolpyruvate carboxylase (ppc) polypeptide, and wherein the transposon insertion down-regulates the ppc gene, and wherein the recombinant host cell further expresses an exogenous polynucleotide encoding a thioesterase, wherein said recombinant host cell produces a fatty acid derivative composition at a higher titer, yield or productivity than a corresponding wild type host cell when cultured in a medium containing a carbon source under conditions effective to express said ppc polypeptide.

2. A cell culture comprising the recombinant host cell according to claim 1 .

3. The cell culture of claim 2 , wherein the fatty acid derivative is
a) a C6, C8, C10, C12, C13, C14, C15, C16, C17, or C18 fatty acid derivative, or
b) a C10:1, C12:1, C14:1, C16:1, or C18:1 unsaturated fatty acid derivative.

4. The cell culture of claim 2 , wherein said recombinant host cell produces a fatty acid derivative yield that is at least about 5% greater than that of said corresponding wild type host cell when cultured under the same conditions as the recombinant host cell.

5. The cell culture of claim 4 , wherein said titer is from about 1 g/L to about 70 g/L.

6. The cell culture of claim 2 , wherein said recombinant host cell produces a fatty acid derivative yield that is at least about 10% to about 40% greater than that of said corresponding wild type host cell when cultured under the same conditions as the recombinant host cell.

7. The cell culture of claim 2 , wherein said productivity ranges from about 0.7 mg/L/hr to about 3 g/L/hr.

8. The cell culture of claim 2 , wherein said cell culture comprises a fatty acid derivative composition.

9. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises at least one fatty acid derivative selected from the group consisting of fatty acid, a fatty ester, a fatty alcohol, a fatty aldehyde, an alkane, a terminal olefin, an internal olefin, and a ketone.

10. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises:
a) one or more of C8, C10, C12, C14, C16, and C18 fatty acid derivatives,
b) fatty acids,
c) fatty aldehydes,
d) fatty alcohols,
e) fatty esters,
f) alkanes,
g) terminal olefins,
h) internal olefins, or
i) ketones.

11. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises fatty acid derivatives having a double bond at position 7 in the carbon chain between C7 and C8 from the reduced end of a fatty alcohol.

12. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises unsaturated fatty acid derivatives.

13. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises saturated fatty acid derivatives.

14. The cell culture of claim 8 , wherein the fatty acid derivative composition comprises branched chain fatty acid derivatives.

Assignees

• Genomatica Inc

Inventors

• Greenfield Derek L
• Schirmer Andreas W
• Clarke Elizabeth J
• Groban Eli S
• Da Costa Bernardo M
• Hu Zhihao
• Holden Kevin
• Helman Noah

Classifications

• C12N9/0006 Primary

  acting on CH-OH groups as donors (1.1) · CPC title

• C12P7/6409

  Fatty acids · CPC title

• C12N9/1288

  Transferases for other substituted phosphate groups (2.7.8) · CPC title

• C12Y207/08007

  Holo-[acyl-carrier-protein] synthase (2.7.8.7) · CPC title

• C12Y103/01009

  Enoyl-[acyl-carrier-protein] reductase (NADH) (1.3.1.9) · CPC title