Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A R.L.
InventorsAdriana Leonora Botes, Alex Van Eck Conradie
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Abstract
This document describes biochemical pathways for producing glutaric acid, 5-aminopentanoic acid, 5-hydroxypentanoic acid 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 cadaverine or 5-aminopentanamide.
Claims54
A method of producing 5-aminopentanoate in a recombinant host, said method comprising:
- enzymatically converting lysine to cadaverine in said recombinant host using a polypeptide having decarboxylase activity, said polypeptide having decarboxylase activity classified under EC 4.1.1.-. and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 1, 16, 17, or 18;
- enzymatically converting cadaverine to 5-aminopentanoate in said recombinant host using a polypeptide having oxidase activity, said polypeptide having oxidase activity classified under EC 1.4.3.21 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 21;
- enzymatically converting lysine to 5-aminopentanamide in said recombinant host using a polypeptide having monooxygenase activity and enzymatically converting 5-aminopentanamide to 5-aminopentanoate in said recombinant host using a polypeptide having amidase activity, said polypeptide having monooxygenase activity classified under EC 1.13.12.2 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 20, said polypeptide having amidase activity classified under EC 3.5.1.30 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 19;
- or
- enzymatically converting cadaverine to 5-aminopentanal in said recombinant host using a polypeptide having ω-transaminase activity and enzymatically converting 5-aminopentanal to 5-aminopentanoate in said recombinant host using a polypeptide having aldehyde dehydrogenase activity, said polypeptide having ω-transaminase activity classified under EC 2.6.1.- and having at least 70% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs. 8 to 13, and said polypeptide having aldehyde dehydrogenase activity is classified under EC 1.2.1.3 or EC 1.2.1.4,
- the method optionally further comprising enzymatically converting 5-aminopentanoate to a product selected from the group consisting of glutaric acid, 5-hydroxypentanoate, and 1,5 pentanediol.
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A recombinant host comprising:
- at least one exogenous nucleic acid encoding a polypeptide having decarboxylase activity and a polypeptide having oxidase activity, said host producing 5-aminopentanoate, said polypeptide having decarboxylase activity classified under EC 4.1.1.- and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 1, 16, 17, or 18, said polypeptide having oxidase activity classified under EC 1.4.3.21 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 21;
- at least one exogenous nucleic acid encoding a polypeptide having monooxygenase activity and a polypeptide having amidase activity, said host producing 5-aminopentanoate, said polypeptide having amidase activity classified under EC 3.5.1.30 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 19, said polypeptide having monooxygenase activity classified under EC 1.13.12.2 and having at least 70% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 20;
- or
- at least one exogenous nucleic acid encoding a polypeptide having ω-transaminase activity and a polypeptide having aldehyde dehydrogenase activity, said host producing 5-aminopentanoate, said polypeptide having ω-transaminase activity classified under EC 2.6.1.- and having at least 70% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs. 8 to 13, and said polypeptide having aldehyde dehydrogenase activity classified under EC 1.2.1.3 or EC 1.2.1.4.
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Dependencies are inferred from the claim text.