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US 11,680,280 B2Granted
Recombinant host cells and methods for the production of isobutyric acid
AssigneeLygos, Inc.
InventorsJeffrey Dietrich, Mario Ouellet, Andrew Jonathan Conley
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Abstract
Methods and materials related to producing isobutyric acid are disclosed. Specifically, isolated nucleic acids, polypeptides, host cells, methods and materials for producing isobutyric by direct microbial fermentation from a carbon source are disclosed.
Claims17
A recombinant host cell, comprising:
- an isobutyric acid biosynthetic pathway, comprising heterologous nucleic acids encoding an acetolactate synthase, a ketol-acid reductoisomerase, a dihydroxy-acid dehydratase, a branched-chain-2-oxoacid decarboxylase, and an isobutyraldehyde dehydrogenase;
- wherein the recombinant host cell is Pichia kudriavzevii host cell,
- wherein the amino acid sequence of the acetolactate synthase comprises SEQ ID NO:
- 1 or SEQ ID NO:
- 19,
- wherein the amino acid sequence of the ketol-acid reductoisomerase comprises SEQ ID NO:
- 2, SEQ ID NO:
- 20, or SEQ ID NO:
- 21,
- wherein the amino acid sequence of the dihydroxy-acid dehydratase comprises SEQ ID NO:
- 3 or SEQ ID NO:
- 22,
- wherein the amino acid sequence of the branched-chain-2-oxoacid decarboxylase comprises SEQ ID NO:
- 4 or SEQ ID NO:
- 23, and
- wherein the amino acid sequence of the isobutyraldehyde dehydrogenase comprises SEQ ID NO:
- 5 or SEQ ID NO:
- 24,
- wherein the heterologous nucleic acids are expressed in sufficient amounts in the recombinant host cell to enable the recombinant host cell to produce increased amounts of isobutyric acid compared to a wild type host cell.
16 more elements
An isobutyric acid biosynthetic pathway, comprising at least one heterologous enzyme selected from the group consisting of:
- (i) an acetolactate synthase that has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% amino acid identity to SEQ ID NO:
- 19;
- (ii) a ketol-acid reductoisomerase that has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% amino acid identity to SEQ ID NO:
- 21;
- (iii) a dihydroxy-acid dehydratase that has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% amino acid identity to SEQ ID NO:
- 22;
- (iv) a branched-chain-2-oxoacid decarboxylase that has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% amino acid identity to SEQ ID NO:
- 23;
- and
- (v) an isobutyraldehyde dehydrogenase that has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% amino acid identity to SEQ ID NO:
- 24.
8 more elements
Dependencies are inferred from the claim text.