Production of fatty acid derivatives
AssigneeReg Life Sciences, LLC
InventorsDerek L. Greenfield, Andreas W. Schirmer, Elizabeth J. Clarke, Eli S. Groban, Bernardo M. Da Costa, Zhihao Hu
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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.
Claims63
A cultured genetically engineered host cell comprising:
- (a) a polynucleotide sequence encoding one or more of:
- (i) an acetyl-CoA carboxylase (EC 6.4.1.2) polypeptide, (ii) a FadR polypeptide, (iii) a heterologous iFAB polypeptide, (iv) a sequence having a transposon insertion in the yijP gene, and (v) a heterologous ACP protein;
- and
- (b) a polynucleotide sequence encoding a fatty acid derivative biosynthetic polypeptide, wherein the genetically engineered host cell produces a fatty acid derivative composition at a higher titer, yield or productivity when cultured in medium containing a carbon source under conditions effective to overexpress the polynucleotide(s) relative to a corresponding wild type host cell propagated under the same conditions as the genetically engineered host cell.
A cultured recombinant host cell, engineered to increase the production of malonyl CoA, comprising:
- a polynucleotide sequence encoding one or more of:
- (i) an acetyl-CoA carboxylase (EC 6.4.1.2) polypeptide, (ii) a FadR polypeptide, (iii) a heterologous iFAB polypeptide, or (iv) a sequence having a transposon insertion in the yijP gene, wherein the engineered host cells produces a fatty acid derivative composition at a higher titer, yield or productivity when cultured in medium containing a carbon source under conditions effective to overexpress the polynucleotide(s) relative to a corresponding wild type host cell propagated under the same conditions as the genetically engineered host cell.
The cultured recombinant host cell of any one of claims 40 to 49, wherein the acetyl-CoA carboxylase (EC 6.4.1.2) polypeptide is overexpressed.
A method of making a fatty acid derivative composition, comprising the steps of:
- (a) engineering a parental host cell to obtain a recombinant host cell which comprises an acetyl-CoA carboxylase (EC 6.4.1.2) polypeptide, (ii) a FadR polypeptide, (iii) a heterologous iFAB polypeptide, and (iv) a sequence having a transposon insertion in the yijP gene;
- (b) further engineering the cell to comprise polynucleotide sequence encoding a fatty acid derivative biosynthetic polypeptide;
- (c) culturing the recombinant host cell in the presence of a carbon source under conditions effective to result in a yield, titer or productivity of the fatty acid derivative composition that is at least 3 times the yield, titer or productivity of fatty acid derivative composition produced by the parental microbial cell cultured under the same conditions;
- and
- (d) optionally isolating the fatty acid derivative composition.
A method of making a fatty acid derivative composition with a higher titer, yield or productivity of fatty acid derivatives than produced by a parental host cell, the method comprising:
- (a) engineering a parental host cell to obtain a recombinant host cell which comprises one or more of:
- (i) a polynucleotide encoding an acetyl-CoA carboxylase (EC 6.4.1.2) polypeptide, (ii) a polynucleotide encoding a FadR polypeptide, (iii) a polynucleotide encoding a heterologous iFAB polypeptide, (iv) a sequence having a transposon insertion in the yijP gene, and (v) a polynucleotide encoding a heterologous ACP protein;
- (b) further engineering the cell to comprise a polynucleotide sequence encoding a fatty acid derivative biosynthetic polypeptide;
- (c) culturing the recombinant host cell in the presence of a carbon source under conditions effective to result in a yield, titer or productivity of fatty acid derivatives that is at least 3 times the yield, titer or productivity of fatty acid derivatives produced by the parental microbial cell cultured under the same conditions;
- and
- (d) optionally isolating the fatty acid derivative composition.
Dependencies are inferred from the claim text.