Genetically modified yeast cells and methods of use for increased lipid yield
AssigneeGinkgo Bioworks, Inc.
InventorsAnnapurna Kamineni, Arthur J. Shaw, Vasiliki Tsakraklides
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Abstract
Aspects of the disclosure are directed to genetically modified yeast cells and methods for use. Certain aspects are directed to recombinant yeast cells comprising exogenous nucleic acid sequences encoding phosphotransacetylase and/or phosphoketolase proteins, including a phosphoketolase protein from Clostridium acetobutylicum. Also disclosed are methods for generating recombinant yeast cells and methods of use of such cells for production of one or more products, including lipids, oils, fatty acids, and triacylglycerides.
Claims161
A recombinant yeast cell, comprising:
- (a) a first exogenous nucleic acid sequence encoding a phosphoketolase protein comprising a sequence having at least 90% sequence identity to SEQ ID NO:28;
- and
- (b) a second exogenous nucleic acid sequence encoding a phosphotransacetylase protein.
The recombinant yeast cell of claim 36, wherein the phosphotransacetylase protein comprises SEQ ID NO:30.
The recombinant yeast cell of any of claims, wherein the recombinant yeast cell has reduced expression of an endogenous functional phosphofructokinase protein compared to a wild type yeast cell.
A method for collecting an acetyl-CoA derived product from a yeast cell, the method comprising:
- (a) culturing a recombinant yeast cell comprising:
- (i) a first exogenous nucleic acid sequence encoding a phosphoketolase protein comprising a sequence having at least 90% sequence identity to SEQ ID NO:28;
- and
- (ii) a second exogenous nucleic acid sequence encoding a phosphotransacetylase protein
- under conditions sufficient to express the first exogenous nucleic acid sequence and the second exogenous nucleic acid sequence;
- and
- (b) collecting the acetyl-CoA derived product from the yeast cell.
A recombinant Yarrowia lipolytica cell, comprising:
- (a) a first exogenous nucleic acid sequence encoding a phosphoketolase protein from Clostridium acetobutylicum, wherein the first exogenous nucleic acid sequence comprises a sequence having at least 95% identity to SEQ ID NO:27;
- and
- (b) a second exogenous nucleic acid sequence encoding (i) a phosphotransacetylase protein from Bacillus subtilis or (ii) a phosphotransacetylase protein from Thermoanaerobacterium saccharolyticum.
Dependencies are inferred from the claim text.