Synthetic biochemistry molecular purge valve module that maintain co-factor balance
AssigneeThe Regents of the Univesity 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.
Claims20
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 of a cofactor cofactors, said pathway comprising:
- a first cofactor-dependent enzyme that is capable of converting a first substrate to a second substrate, said first cofactor-dependent enzyme producing the unbalanced production of a first cofactor;
- and
- a second cofactor-dependent enzyme comprising that uses a second cofactor different from the first cofactor-dependent enzyme and that converts the first substrate to the second substrate and wherein said second cofactor-dependent enzyme produces an unbalanced production of the second cofactor, and wherein the pathway comprises an NADH or an NADPH oxidase that recycles the an NADH or NADPH 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 first cofactor-dependent enzyme that is capable of converting the substrate to the product, said first cofactor-dependent enzyme producing the unbalanced production of a first cofactor;
- and
- a second cofactor-dependent enzyme that uses a second cofactor than the first cofactor-dependent enzyme and that converts the substrate to the product and wherein said second cofactor-dependent enzyme produces an unbalanced production of the second cofactor, and a metabolic purge valve comprising an NADH pyruvate dehydrogenase and a NADH/NADPH an NADH or an NADPH oxidase.
A recombinant polypeptide comprising a sequence that has at least 99% 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.