Glycerol 3- phosphate dehydrogenase for butanol production
First Claim
1. A recombinant microorganism comprising:
- (a) an isobutanol biosynthetic pathway comprising at least one polypeptide that is heterologous to the recombinant microorganism;
(b) a heterologous glycerol-3-phosphate dehydrogenase (GPD), wherein the heterologous GPD has at least 90% identity to SEQ ID NO;
195 and comprises at least one substitution at a residue corresponding to position 44, 45, 73, or 129 of SEQ ID NO;
195, andwherein the heterologous GPD has a higher KM for NADH as compared to the KM of the endogenous GPD of the microorganism; and
(c) a deletion or disruption in an endogenous gene encoding GPD;
wherein the recombinant microorganism has improved production of isobutanol as compared to a control recombinant microorganism that lacks the heterologous GPD.
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Accused Products
Abstract
Provided herein are glycerol-3-phosphate dehydrogenase (GPD) enzymes with increased KM for NADH and GPD enzymes with substantially the same affinity for NADH and NADPH and/or are feedback inhibited by glycerol-3-phosphate. Also provided herein are recombinant microorganisms comprising a heterologous gene encoding GPD and a deletion or disruption in an endogenous gene encoding GPD. Also provided are recombinant microorganisms comprising a heterologous gene encoding GPD and a butanol biosynthetic pathway. Further provided are methods of producing butanol comprising providing the recombinant microorganisms described herein and contacting the recombinant microorganism with at least one fermentable carbon substrate under conditions wherein butanol is produced.
114 Citations
8 Claims
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1. A recombinant microorganism comprising:
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(a) an isobutanol biosynthetic pathway comprising at least one polypeptide that is heterologous to the recombinant microorganism; (b) a heterologous glycerol-3-phosphate dehydrogenase (GPD), wherein the heterologous GPD has at least 90% identity to SEQ ID NO;
195 and comprises at least one substitution at a residue corresponding to position 44, 45, 73, or 129 of SEQ ID NO;
195, andwherein the heterologous GPD has a higher KM for NADH as compared to the KM of the endogenous GPD of the microorganism; and (c) a deletion or disruption in an endogenous gene encoding GPD; wherein the recombinant microorganism has improved production of isobutanol as compared to a control recombinant microorganism that lacks the heterologous GPD. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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Specification