Methods, reagents and cells for biosynthesizing glutarate methyl ester
AssigneeInvista North America S.A.R.L.
InventorsNadia Fatma Kadi, Mariusz Kamionka, Alexander Brett Foster, Alex Van Eck Conradie, Adriana Leonora Botes
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Abstract
This document describes biochemical pathways for producing 2,4-pentadienoyl-CoA by forming one or two terminal functional groups, comprised of carboxyl or hydroxyl group, in a C5 backbone substrate such as glutaryl-CoA, glutaryl-[acp] or glutarate methyl ester. 2,4-pentadienoyl-CoA can be enzymatically converted to 1,3-butadiene.
Claims20
A method of biosynthesizing glutarate methyl ester in a recombinant host comprising at least one exogenous nucleic acid encoding a polypeptide having malonyl-CoA O-methyltransferase activity, the method comprising enzymatically converting at least one of malonyl-[acp] and malonyl-CoA to glutarate methyl ester in said host using a polypeptide having malonyl-CoA O-methyltransferase activity, a polypeptide having thioesterase activity, or a combination thereof, wherein said polypeptide having malonyl-CoA O-methyltransferase activity has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:13 and is capable of enzymatically converting malonyl-CoA to malonyl-CoA methyl ester or malonyl-[acp] to malonyl-[acp] methyl ester, and wherein said polypeptide having thioesterase activity has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:14, or SEQ ID NO:15 and is capable of enzymatically converting glutaryl-CoA methyl ester to glutarate methyl ester,
- wherein malonyl-CoA is enzymatically converted to malonyl-CoA methyl ester using said at least one polypeptide having malonyl-CoA O-methyltransferase activity;
- or
- wherein malonyl-[acp] is enzymatically converted to malonyl-[acp] methyl ester using said at least one polypeptide having malonyl-CoA O-methyltransferase activity,
- the method optionally further comprising enzymatically converting glutarate methyl ester to glutarate semialdehyde methyl ester in said host using at least one polypeptide having carboxylate reductase activity, wherein said polypeptide having carboxylate reductase activity has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 or SEQ ID NO:7 and is capable of enzymatically converting glutarate methyl ester to glutarate semialdehyde methyl ester,
- wherein said enzymatic conversion occurs in the host using said polypeptide having malonyl-CoA O-methyltransferase activity, said polypeptide having thioesterase activity, or a combination thereof.
Dependencies are inferred from the claim text.