Synthetic biochemistry molecular purge valve module that maintain co-factor balance
AssigneeThe Regents of the University of California
InventorsJames U. Bowie, Paul H. Opgenorth, Tyler P. Korman
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Abstract
The disclosure provides a metabolic pathway for producing a metabolite, the metabolic pathway having a co-factor purge valve system for recycling a cofactor used in the metabolic pathway.
Claims18
A recombinant, artificial or engineered metabolic pathway comprising a plurality of enzymatic steps that converts a substrate to a product, wherein the pathway produces an unbalanced production and utilization of a cofactor, said pathway comprising:
- a first cofactor-dependent enzyme that is capable of converting a first substrate to a second substrate, said enzyme producing the unbalanced production and utilization of a cofactor;
- a second cofactor-dependent enzyme that is also capable of converting the first substrate to a second substrate, wherein the second cofactor-dependent enzyme is the first cofactor dependent enzyme having its cofactor preference altered;
- and
- an enzyme that recycles the cofactor.
An enzymatic system comprising a metabolic pathway including a plurality of enzymes for converting a substrate to a product, the metabolic pathway having an unbalanced utilization of reducing/oxidizing cofactors, wherein the enzymatic system comprises a metabolic purge valve comprising an NADH pyruvate dehydrogenase, and NADPH pyruvate dehydrogenase and a NADH/NADPH oxidase.
A recombinant polypeptide comprising a sequence that has at least 90% sequence identity to SEQ ID NO:
- 5 and comprising the mutations E206V, G207R, A208K, and S213R, wherein the polypeptide has dihydrolipoamide dehydrogenase activity.
Dependencies are inferred from the claim text.