Combinatorial Metabolic Engineering Of Saccharomyces Cerevisiae For Terminal Alkene Production
AssigneeNational University of Singapore
InventorsMatthew Wook Chang, Binbin Chen
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Abstract
Modified Saccharomyces cerevisiae yeast that produce terminal alkenes are described. The modification of the Saccharomyces cerevisiae yeast includes insertion of at least one heterologous fatty acid decarboxylase gene, deletion of FAA1 and FAA4, overexpression of HEM3, and triple-deletion of CTT1, CTA1 and CCP1. Methods of producing terminal alkenes by culturing and fermenting the modified Saccharomyces cerevisiae yeast and optionally harvesting the terminal alkenes are also described. Mixtures of terminal alkenes produced by the modified Saccharomyces cerevisiae yeast, and methods of metabolically engineering a yeast for optimizing overexpression of one or more alkenes are also described.
Claims20
A modified Saccharomyces cerevisiae yeast wherein the modification comprises:
- insertion of at least one heterologous fatty acid decarboxylase gene,
- deletion of FAA1 and FAA4,
- overexpression of HEM3, and
- triple-deletion of CTT1, CTA1 and CCP1.
A method of metabolically engineering a yeast for optimizing overexpression of one or more alkenes, the method comprising:
- a) selecting a yeast having inserted therein one or more heterologous decarboxylase genes for alkene biosynthesis in the yeast via free fatty acid decarboxylation;
- b) enhancing the metabolic flux towards free fatty acid production in the yeast by disrupting the fatty acid metabolic pathway by deleting at least one synthetase and optionally overexpressing at least one carboxylase;
- c) supplying at least one decarboxylase cofactor to the alkene biosynthesis pathway to enhance the metabolic flux towards alkene production in the yeast;
- d) tuning expression levels of the one or more heterologous decarboxylase genes by at least one of promoter strength tuning, plasmid copy number tuning and growth medium tuning;
- and
- e) optimizing yeast fermentation conditions by at least one of temperature control, dissolved oxygen concentration control and pH control,
- thereby optimizing overexpression of the one or more alkenes by the metabolically engineered yeast.
Dependencies are inferred from the claim text.