Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A R.L.
InventorsAdriana Leonora Botes, Alex Van Eck Conradie
Life of this patent
Estimating the term…
Abstract
This document describes biochemical pathways for producing glutaric acid, 5-aminopentanoic acid, 5-hydroxypentanoic acid, cadaverine or 1,5-pentanediol by forming one or two terminal functional groups, comprised of carboxyl, amine or hydroxyl group, in a C5 backbone substrate such as malonyl-CoA or malonyl-[acp].
Claims71
A method of biosynthesizing glutarate methyl ester in a recombinant host, the method comprising enzymatically converting at least one of malonyl-[acp] and malonyl-CoA to glutarate methyl ester in said host, wherein said enzymatic conversion occurs in the host using a polypeptide having malonyl-CoA O-methyltransferase activity, a polypeptide having thioesterase activity, or a combination thereof;
- wherein malonyl-[acp] is enzymatically converted to malonyl-[acp] methyl ester using said at least one polypeptide having malonyl-CoA O-methyltransferase activity, wherein said polypeptide having malonyl-CoA O-methyltransferase activity has at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 21,
- 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 the polypeptide having carboxylate reductase activity has at least 70% sequence identity to any one of the amino acid sequences set forth in any one of SEQ ID NOs:
- 2-7.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(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, wherein said polypeptide having pimeloyl-[acp] methyl ester methylesterase activity has at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 1, and (ii) enzymatically converting glutaryl-[acp] or glutaryl-CoA to glutaric acid 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.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(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, wherein the polypeptide having thioesterase activity has at least 70% sequence identity to any one of the amino acid sequences set forth in any one of SEQ ID NOs:
- 17-18 and wherein the polypeptide having malonyl-CoA O-methyltransferase activity has at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 21,
- the host optionally further comprising an exogenous polypeptide having carboxylate reductase activity, wherein the polypeptide having reductase activity has at least 70% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOs:
- 19 or 20, said host further producing glutarate semialdehyde methyl ester.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
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, wherein said polypeptide having pimeloyl-[acp] methyl ester methylesterase activity has at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 1.
(canceled)
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,
- the method optionally further comprising converting said HC(═O)(C2-C6 alkyl)-C(═O)OCH3 to a HOCH2(C2-C6 alkyl)-C(═O)OCH3 using a polypeptide having dehydrogenase activity.
(canceled)
The method of claim 71, further comprising enzymatically converting the HOCH2(C2-C6 alkyl)-C(═O)OCH3 to a HOCH2(C2-C6 alkyl)-C(═O)OH using a polypeptide having esterase activity.
Dependencies are inferred from the claim text.