Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A.R.L.
InventorsAlex Van Eck Conradie, Adriana Leonora Botes
Life of this patent
Estimating the term…
Abstract
This document describes biochemical pathways for producing 2(E)-heptenedioyl-CoA methyl ester from precursors such as 2-oxo-glutarate, acetyl-CoA, or succinyl-CoA using one or more of a fatty acid O-methyltransferase, a thioesterase, a CoA-transferase, a CoA ligase, as well as recombinant hosts expressing one or more of such enzymes. 2(E)-heptenedioyl-CoA methyl ester can be enzymatically converted to pimeloyl-CoA using a trans-2-enoyl-CoA reductase, and a methylesterase. Pimeloyl-CoA can be enzymatically converted to pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, or 1,7-heptanediol.
Claims30
A method of shielding a carbon chain aliphatic backbone, functionalized with terminal carboxyl groups, in a recombinant host using at least one exogenous polypeptide expressed by said host, said method comprising:
- enzymatically converting a n-carboxy-2-enoic acid to a n-carboxy-2-enoate methyl ester in said host by contacting said n-carboxy-2-enoic acid with a polypeptide having fatty acid O-methyltransferase activity, wherein n+1 reflects the length of the carbon chain aliphatic backbone and the polypeptide having fatty acid O-methyltransferase activity has at least 85% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:
- 1-3.
A method of producing 2(E)-heptenedioyl-CoA methyl ester in a recombinant host using at least one exogenous polypeptide expressed by said host, said method comprising:
- enzymatically converting 2(E)-heptenedioate to 2(E)-heptenedioate methyl ester by contacting 2(E)-heptenedioate with a polypeptide having fatty acid O-methyltransferase activity, wherein said polypeptide having fatty acid O-methyltransferase activity has at least 85% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:
- 1-3;
- enzymatically converting 2(E)-heptenedioate methyl ester to 2(E)-heptenedioyl-CoA methyl ester by contacting 2(E)-heptenedioate methyl ester with a polypeptide having the activity of a CoA ligase, wherein said polvpeptide having the activity of a CoA ligase has at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 22 or SEQ ID NO:
- 23;
- enzymatically converting 2(E)-heptenedioyl-CoA methyl ester to pimeloyl-CoA methyl ester by contacting 2(E)-heptenedioyl-CoA methyl ester with a polypeptide having the activity of a trans-2-enoyl-CoA reductase, wherein said polvpeptide having the activity of a trans-2-enoyl-CoA reductase has at least 85% sequence identity to the amino acid sequence of SEQ ID NO:
- 26 or SEQ ID NO:
- 27;
- and
- enzymatically converting pimeloyl-CoA methyl ester to pimeloyl-CoA by contacting pimeloyl-CoA methyl ester with a polypeptide having the activity of a pimelyl-[acp]methyl ester esterase, wherein said polypeptide having the activity of a pimelyl-[acp]methyl ester esterase has at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 6.
A method of shielding a carbon chain aliphatic backbone, functionalized with terminal carboxyl groups, in a recombinant host using at least one exogenous polypeptide expressed by said host, said method comprising:
- enzymatically converting a n-carboxy-2-enoic acid to a n-carboxy-2-enoate methyl ester in said host by contacting said n-carboxy-2-enoic acid with a polypeptide having at least 85% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:
- 1-3, wherein n+1 reflects the length of the carbon chain aliphatic backbone.
A method of producing 2(E)-heptenedioyl-CoA methyl ester in a recombinant host using at least one exogenous polypeptide expressed by said host, said method comprising:
- enzymatically converting 2(E)-heptenedioate to 2(E)-heptenedioate methyl ester by contacting 2(E)-heptenedioate with a polypeptide having at least 85% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:
- 1-3;
- enzymatically converting 2(E)-heptenedioate methyl ester to 2(E)-heptenedioyl-CoA methyl ester by contacting 2(E)-heptenedioate methyl ester with a polypeptide having at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 22 or SEQ ID NO:
- 23;
- enzymatically converting 2(E)-heptenedioyl-CoA methyl ester to pimeloyl-CoA methyl ester by contacting 2(E)-heptenedioyl-CoA methyl ester with a polypeptide having at least 85% sequence identity to the amino acid sequence of SEQ ID NO:
- 26 or SEQ ID NO:
- 27;
- and
- enzymatically converting pimeloyl-CoA methyl ester to pimeloyl-CoA by contacting pimeloyl-CoA methyl ester with a polypeptide having at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 6.
Dependencies are inferred from the claim text.