Granted Patent
B2
US 12,595,466 · App. 17/909,353 · Granted Apr 7, 2026
NKR variants for increased production of isobutanol
Inventors:
Christopher Smith (Englewood, CO); Timothy Hanly (Englewood, CO); Jeanne Benoit (Englewood, CO)
Assignee:
GEVO, INC.
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Abstract
The present invention relates to recombinant microorganisms comprising at least one nucleic acid molecule encoding a ketol-acid reductoisomerase (KARI) or modified NADH-dependent variant thereof, wherein said KARI is at least about 60% identical to SEQ ID NO: 2. In various aspects of the invention, the recombinant microorganisms may comprise an isobutanol producing metabolic pathway and can be used in methods of making isobutanol.
Claims (12)
1 . A recombinant microorganism comprising at least one nucleic acid molecule encoding a ketol-acid reductoisomerase (KARI), wherein said KARI is at least about 98% identical to SEQ ID NO: 90, SEQ ID NO: 92, or SEQ ID NO: 94.
2 . The recombinant microorganism of claim 1 , wherein said KARI is derived from the genus Slackia.
3 . The recombinant microorganism of claim 2 , wherein said KARI is derived from Slackia exigua.
4 . The recombinant microorganism of claim 1 , wherein said KARI comprises the amino acid sequence of SEQ ID NO: 90, SEQ ID NO: 92, or SEQ ID NO: 94.
5 . The recombinant microorganism of claim 1 , wherein said KARI is encoded by SEQ ID NO:89, SEQ ID NO: 91, or SEQ ID NO: 93.
6 . The recombinant microorganism of claim 1 , wherein said KARI is NADH-dependent.
7 . The recombinant microorganism of claim 1 , wherein said recombinant microorganism further comprises an isobutanol producing metabolic pathway comprising one or more isobutanol metabolic pathway enzymes selected from acetolactate synthase, dihydroxy acid dehydratase, keto-isovalerate decarboxylase, and alcohol dehydrogenase.
8 . The recombinant microorganism of claim 1 , wherein said recombinant microorganism comprises a metabolic pathway for the production of a metabolite selected from the group consisting of isoleucine, leucine, valine, pantothenate, coenzyme A, 1-butanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 3-methyl-1-pentanol, 4-methyl-1-pentanol, 4-methyl-1-hexanol, and 5-methyl-1-heptanol.
9 . The recombinant microorganism of claim 1 , wherein said recombinant microorganism is a yeast microorganism.
10 . A method of producing isobutanol, comprising:
a. providing a recombinant microorganism of claim 1 ; and
b. cultivating the recombinant microorganism in a culture medium containing a feedstock providing a carbon source, until the isobutanol is produced.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 17, 2023
From: SMITH, CHRISTOPHER; HANLY, TIMOTHY; BENOIT, JEANNE
To: GEVO, INC.
Reel/Frame 062397/0775 →
Continuity (2)
Provisional Application
62986378
· Mar 6, 2020
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