Recycling system for manipulation of intracellular NADH availability
AssigneeRice University
InventorsKa-Yiu San, Susana J. Berrios-Rivera, George N. Bennett
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Abstract
The present invention describes a novel recombinant NADH recycling system that is used as a process for producing reduced compounds. In a specific embodiment, the reduced compounds include ethanol, succinate, lactate, a vitamin, a pharmaceutical and a biodegraded organic molecule. The NADH recycling system effects metabolic flux of reductive pathways in aerobic and anaerobic environments.
Claims8
An isolated engineered cell, comprising a recombinant NADH-recycling system having:
- (a) a native cofactor-independent formate dehydrogenase genetically engineered to reduce its enzymatic activity as compared with the activity in a non-engineered cell, and
- (b) an expression construct comprising an expressible gene encoding a heterologous NAD+-dependent formate dehydrogenase (FDH) that increases the enzymatic activity of NAD+-dependent FDH in the cell as compared with the activity of NAD+-dependent FDH in a non-engineered cell,
- wherein said FDH converts NAD+ to NADH in the presence of formate.
An isolated engineered bacterial cell with a recombinant NADH-recycling system comprising:
- (a) a native cofactor-independent formate dehydrogenase that has been inactivated by mutation or deletion, and
- (b) an expression construct comprising an expressible gene encoding a heterologous NAD+-dependent formate dehydrogenase (FDH) that results in overexpression of the NAD+-dependent FDH as compared with the expression of NAD+-dependent FDH in a non-engineered cell, wherein said FDH converts NAD+ to NADH in the presence of formate.
A bacterial cell comprising an expression vector encoding a heterologous NAD+-dependent formate dehydrogenase (FDH) and lacking cofactor-independent formate dehydrogenase activity, wherein said FDH converts NAD+ to NADH in the presence of formate.
Dependencies are inferred from the claim text.