Yeast cell modified to overproduce fatty acid and fatty acid-derived compounds
AssigneeThe Regents of the University of California
InventorsWeerawat Runguphan, Jay D. Keasling
Life of this patent
Estimating the term…
Abstract
The present invention provides for a genetically modified yeast host cell capable of producing one or more fatty acids, or fatty acid-derived compounds, or a mixture thereof, comprising: (a) increased expression of acetyl-CoA carboxylase (such as ACC1), (b) increased expression of one or more fatty acid synthases (such as FAS1 and FAS2), and (c) optionally reduced expression of one or more enzymes involved in or in the β-oxidation pathway (such as peroxisomal transporters PXA1 and PXA2, and β-oxidation enzymes POX1, POX2, and POX3).
Claims11
A genetically modified Saccharomyces cerevisiae host cell capable of producing one or more fatty acids, or fatty acid-derived compounds, or a mixture thereof, comprising:
- (a) increased expression of acetyl-CoA carboxylase, wherein the endogenous promoter of the ACC1 gene is replaced with promoter PTEF1;
- (b) increased expression of one or more fatty acid synthases, wherein the endogenous promoter of the FAS1 and FAS2 genes are replaced with promoter PTEF1;
- (c) optionally reduced expression of one or more enzymes in the β-oxidation pathway, wherein the endogenous POX1 and/or PXA2 genes are deleted;
- (d) a plasmid encoding an Escherichia coli TesA, which lacks a membrane signal peptide at a N-terminal end of TesA, operatively linked to promoter PGAL10;
- and
- (e) reduced expression of fatty-acyl-CoA synthetases, wherein the endogenous FAA1 and/or FAA4 genes are deleted;
- wherein the host cell does not comprise the ΔSnf2 mutation and the host cell is capable of producing at least 162 mg of free fatty acids when cultured in one liter of culture medium.
A method of constructing a genetically modified Saccharomyces cerevisiae host cell of the claimed invention, comprising:
- (a) replacing the endogenous promoter of the ACC1 gene with promoter PTEF1 and the endogenous promoter of the FAS1 and FAS2 genes with promoter PTEF1;
- (b) optionally the deleting the endogenous POX1 and/or PXA2 genes;
- (c) introducing a plasmid encoding an Escherichia coli TesA, which lacks a membrane signal peptide at a N-terminal end of TesA, operatively linked to promoter PGAL10 into the host cell;
- (d) deleting the endogenous FAA1 and/or FAA4 genes;
- and
- (e) culturing the host cell in a cell culture such that the cell culture produces at least 162 mg of free fatty acids per one liter of culture medium;
- wherein the host cell does not comprise the ΔSnf2 mutation.
Dependencies are inferred from the claim text.