Back to results
US 12,644,139 B2Granted
Cells and methods for producing methyl ketones
AssigneeWisconsin Alumni Research Foundation
InventorsBrian F. Pfleger, Qiang Yan, Trevor R. Simmons, Nestor Hernandez-Lozada
Life of this patent
Estimating the term…
Abstract
Recombinant cells and methods for producing methyl ketones, such as medium-chain methyl ketones. The recombinant cells include recombinant acyl-ACP thioesterase genes, recombinant β-ketoacyl-CoA thioesterase genes, and recombinant acyl-CoA synthetase genes, in addition to other modifications. The methods include culturing the recombinant cells to produce the methyl ketones and isolating the produced methyl ketones.
Claims11
A recombinant cell for producing methyl ketones comprising:
- a recombinant β-ketoacyl-CoA thioesterase gene;
- a recombinant acyl-ACP thioesterase gene;
- a recombinant acyl-CoA synthetase gene;
- a recombinant 3-ketoacyl-CoA thiolase gene;
- and
- a functional deletion of at least one of a native enoyl-CoA hydratase gene and a native 3-hydroxyacyl-CoA dehydrogenase gene.
3 more elements
A recombinant cell for producing methyl ketones comprising:
- a recombinant β-ketoacyl-CoA thioesterase gene, wherein the recombinant β-ketoacyl-CoA thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:8;
- a recombinant acyl-ACP thioesterase gene, wherein the recombinant acyl-ACP thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:2;
- a recombinant acyl-CoA synthetase gene, wherein the recombinant acyl-CoA synthetase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:12;
- a functional deletion of a native acyl-CoA dehydrogenase gene;
- a functional deletion of at least one of a native enoyl-CoA hydratase gene and a native 3-hydroxyacyl-CoA dehydrogenase gene;
- and
- at least one of:
- a functional deletion of a native fadR gene or a homolog thereof;
- and
- a recombinant 3-ketoacyl-CoA thiolase gene that encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:
- 22 or SEQ ID NO:242,
- wherein the recombinant cell exhibits enhanced production of 2-nonanone with respect to a corresponding native cell.
9 more elements
A recombinant cell for producing methyl ketones comprising:
- a recombinant β-ketoacyl-CoA thioesterase gene, wherein the recombinant β-ketoacyl-CoA thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:8;
- a recombinant acyl-ACP thioesterase gene, wherein the acyl-ACP thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:
- 2;
- a recombinant acyl-CoA synthetase gene, wherein the recombinant acyl-CoA synthetase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:12;
- a functional deletion of a native acyl-CoA dehydrogenase gene;
- a recombinant acyl-CoA dehydrogenase gene that encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:30;
- a functional deletion of a native 3-ketoacyl-CoA thiolase gene;
- and
- at least one of:
- a functional deletion of a native fadR gene or a homolog thereof;
- a recombinant enoyl-CoA hydratase gene;
- and
- a recombinant 3-hydroxyacyl-CoA dehydrogenase gene,
- wherein the recombinant cell exhibits enhanced production of 2-heptanone with respect to a corresponding native cell.
11 more elements
A recombinant cell for producing methyl ketones comprising:
- a recombinant β-ketoacyl-CoA thioesterase gene, wherein the recombinant β-ketoacyl-CoA thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:8;
- a recombinant acyl-ACP thioesterase gene, wherein the acyl-ACP thioesterase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:
- 4;
- a recombinant acyl-CoA synthetase gene, wherein the recombinant acyl-CoA synthetase gene encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:10;
- a functional deletion of a native acyl-CoA dehydrogenase gene;
- a recombinant acyl-CoA dehydrogenase gene that encodes a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:30;
- a functional deletion of a native 3-ketoacyl-CoA thiolase gene;
- and
- at least one of:
- a functional deletion of a native fadR gene or a homolog thereof;
- a recombinant enoyl-CoA hydratase gene encoding a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:16;
- and
- a recombinant 3-hydroxyacyl-CoA dehydrogenase gene encoding a protein comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO:
- 16,
- wherein the recombinant cell exhibits enhanced production of 2-undecanone with respect to a corresponding native cell.
12 more elements
Dependencies are inferred from the claim text.