Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A.R.L.
InventorsNadia Fatma KADI, Mariusz Stanislaw 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.
Claims37
1-22. (canceled)
A method of making glutarate, said method comprising (i) enzymatically converting glutaryl-[acp] methyl ester to glutaryl-[acp] or glutaryl-CoA methyl ester to glutaryl-CoA using a polypeptide having pimeloyl-[acp] methyl ester methylesterase activity, and (ii) enzymatically converting glutaryl-[acp] or glutaryl-CoA to glutarate using at least one polypeptide having thioesterase activity, reversible CoA-ligase activity, a CoA-transferase activity, an acylating dehydrogenase activity, an aldehyde dehydrogenase activity, a glutarate semialdehyde dehydrogenase activity, or a succinate-semialdehyde dehydrogenase activity.
29-30. (canceled)
32-34. (canceled)
(canceled)
38-39. (canceled)
A recombinant host cell comprising at least one exogenous nucleic acid encoding a polypeptide having malonyl-CoA O-methyltransferase activity;
- and a polypeptide having thioesterase activity, the host producing glutarate methyl ester.
A recombinant host comprising at least one exogenous nucleic acid encoding a polypeptide having pimeloyl-[acp] methyl ester methylesterase activity, and at least one polypeptide having an activity selected from the group consisting of thioesterase activity, reversible CoA-ligase activity, a CoA-transferase activity, an acylating dehydrogenase activity, an aldehyde dehydrogenase activity, a glutarate semialdehyde dehydrogenase activity, and a succinate-semialdehyde dehydrogenase activity.
A method of increasing the activity of a polypeptide having carboxylate reductase activity on a substituted or unsubstituted C4-C8 dicarboxylic acid, the method comprising enzymatically converting said C4-C8 dicarboxylic acid to a HOC(═O)(C2-C6 alkyl)-C(═O)OCH3 ester using a polypeptide having malonyl-CoA methyltransferase activity before enzymatically converting the HOC(═O)(C2-C6 alkyl)-C(═O)OCH3 ester to a HC(═O)(C2-C6 alkyl)-C(═O)OCH3 using a polypeptide having carboxylate reductase activity.
Dependencies are inferred from the claim text.