Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A.R.L.
InventorsAlex Van Eck Conradie, Adriana Leonora Botes
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Abstract
This document describes biochemical pathways for producing 2,3-dehydroadipyl-CoA methyl ester from precursors such as 2-oxoglutarate using one or more of a fatty acid O-methyltransferase, a thioesterase, a CoA-transferase and a CoA ligase, as well as recombinant hosts expressing one or more of such enzymes. 2,3-dehydroadipyl-CoA methyl ester can be enzymatically converted to adipyl-CoA using a trans-2-enoyl-CoA reductase, and a methylesterase, which in turn can be enzymatically converted to adipic acid, 6-aminohexanoate, 6-hydroxyhexanoate, caprolactam, hexamethylenediamine, or 1,6-hexanediol.
Claims21
A method of shielding a carbon chain aliphatic backbone functionalized with terminal carboxyl groups in a recombinant host, said method comprising enzymatically converting 2,3-dehydroadipic acid to 2,3-dehydroadipate methyl ester in said host using a polypeptide having the activity of a fatty acid O-methyltransferase, wherein said polypeptide having the activity of a fatty acid O-methyltransferase has at least 85% sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs:
- 1 to 3.
A method of producing 2,3-dehydroadipyl-CoA methyl ester in a recombinant host, said method comprising enzymatically converting 2,3-dehydroadipic acid to 2,3-dehydroadipate methyl ester using a polypeptide having fatty acid O-methyltransferase activity, wherein said polypeptide having fatty acid O-methyltransferase has at least 85% sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs:
- 1 to 3, and further enzymatically converting 2,3-dehydroadipate methyl ester to 2,3-dehydroadipyl-CoA methyl ester using a polypeptide having the activity of a CoA ligase, wherein said polypeptide having the activity of a CoA ligase has at least 85% sequence identity to SEQ ID NO:
- 22 or SEQ ID NO:
- 23,
- wherein the 2,3-dehydroadipic acid is enzymatically produced from 2,3-dehydroadipyl-CoA using a polypeptide having thioesterase activity or a protein having CoA-transferase activity, wherein said polypeptide having thioesterase activity has at least 85% sequence identity to an amino acid sequence set forth in any one of SEQ ID NOs:
- 4, 5, or 21 or at least 85% sequence identity to an amino acid sequence encoding a gene product of yciA or ACOT13 and said protein having CoA-transferase activity has a first subunit having at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 24 and a second subunit having at least 85% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 25,
- the method optionally further comprising enzymatically converting 2,3-dehydroadipyl methyl ester to adipyl-CoA methyl ester using a polypeptide having trans-2-enoyl-CoA reductase activity, wherein said polypeptide having trans-2-enoyl-CoA reductase activity has at least 85% sequence identity to an amino acid sequence set forth in SEQ ID NO:
- 26 or SEQ ID NO:
- 27.
Dependencies are inferred from the claim text.