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US 10,184,139 B2Granted
Integration of a polynucleotide encoding a polypeptide that catalyzes pyruvate to acetolactate conversion
AssigneeButamax Advanced Biofuels LLC
InventorsLarry Cameron Anthony, Lori Ann Maggio-Hall
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Abstract
The invention relates to recombinant host cells having at least one integrated polynucleotide encoding a polypeptide that catalyzes a step in a pyruvate-utilizing biosynthetic pathway, e.g., pyruvate to acetolactate conversion. The invention also relates to methods of increasing the biosynthetic production of isobutanol, 2,3-butanediol, 2-butanol or 2-butanone using such host cells.
Claims15
A recombinant host cell comprising:
- (a) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of pyruvate to acetolactate wherein the polypeptide is an acetolactate synthase from Bacillus subtilis, Klebsiella pneumonia, Lactococcus lactis, Staphylococcus aureus, Listeria monocytogenes, Streptococcus mutans, Streptococcus thermophiles, Vibrio angustum, or Bacillus cereus;
- (b) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of acetolactate to 2,3-dihydroxyisovalerate wherein the polypeptide is a ketol-acid reductoisomerase and the ketol-acid reductoisomerase has at least 95% identity to SEQ ID NO:
- 83;
- (c) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of 2,3-dihydroxyisovalerate to α-ketoisovalerate wherein the polypeptide is a dihydroxyacid dehydratase from Escherichia coli, Bacillus subtilis, Methanococcus maripaludis, or Streptococcus mutans;
- (d) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion of α-ketoisovalerate to isobutyraldehyde wherein the polypeptide is a branched-chain α-keto acid decarboxylase and the branched-chain α-keto acid decarboxylase has at least 95% identity to SEQ ID NO:
- 247;
- and
- (e) a polynucleotide encoding a polypeptide which catalyzes the substrate to product conversion isobutyraldehyde to isobutanol wherein the polypeptide is an alcohol dehydrogenase from Achromobacter xylosoxidans or Beijerinkia indica,
- wherein expression of pyruvate decarboxylase in the recombinant host cell is decreased or eliminated compared to control host.
6 more elements
Dependencies are inferred from the claim text.