Increasing intracellular NADPH availability in E. coli
AssigneeRice University
InventorsKa-Yiu San, George N. Bennett, Ailen Sanchez
Life of this patent
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Abstract
A method of increasing cellular NADPH levels by expressing one or more genes that encode a enzyme that causes the production of NADPH. The system can be combined with other enzymes that require NADPH, thus improving the overall production of the desired protein.
Claims12
A method of increasing the NADPH levels in a bacterial cell, comprising
- a) providing a bacterial host cell,
- b) transforming the host cell with an expression vector encoding a pyridine nucleotide transhydrogenase that catalyzes the following reaction:
- and,
- c) transforming the host cell with one or more genes that encode at least one enzyme that uses NADPH as a cofactor,
- wherein the bacterial host cell produces at least 50% more NADPH after transformation than it did prior to transformation.
A genetically engineered cell that produces more NADPH than the same cell prior to genetic engineering, wherein said cell comprises one or two expression vectors encoding:
- a) a pyridine nucleotide transhydrogenase;
- and
- b) phb operon including β-ketothiolase, acetoacetyl-CoA reductase and PHB synthase, wherein said cell increases NADPH production as measured by increased PHB synthesis.
A genetically engineered bacterial cell, comprising overexpression of pyridine nucleotide transhydrogenase so that at least 50% more NADPH is expressed in said cell as compared with the same cell prior to genetic engineering;
- and overexpression of one or more genes that encode at least one enzyme that uses NADPH as a cofactor.
Dependencies are inferred from the claim text.