Methods, reagents and cells for biosynthesizing compounds
AssigneeInvista North America S.A.R.L.
InventorsAlex Van Eck Conradie, Adriana Leonora Botes, Ramdane Haddouche
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Abstract
This document describes biochemical pathways for producing 6-hydroxyhexanoate methyl ester and hexanoic acid hexyl ester using one or more of a fatty acid O-methyltransferase, an alcohol O-acetyltransferase and a monooxygenase, as well as recombinant hosts expressing one or more of such enzymes. 6-hydroxyhexanoate methyl esters and hexanoic acid hexyl ester can be enzymatically converted to adipic acid, adipate semialdehyde, 6-aminohexanoate, 6-hydroxyhexanoate, hexamethylenediamine, and 1,6-hexanediol.
Claims19
A method of producing a terminal hydroxyl (C3-8 hydroxyalkyl)-C(═O)OCH3 ester in a recombinant host, said method comprising:
- a) enzymatically converting a C4-9 carboxylic acid to a (C3-8 alkyl)-C(═O)OCH3 ester using a polypeptide having fatty acid O-methyltransferase activity classified under EC 2.1.1.15;
- and
- b) enzymatically converting the (C3-8 alkyl)-C(═O)OCH3 ester to a terminal hydroxyl (C3-8 hydroxyalkyl)-C(═O)OCH3 ester using a polypeptide having monooxygenase activity classified under EC 1.14.14.- or EC 1.14.15.-,
- wherein the C4-9 carboxylic acid is enzymatically produced from a C4-9 alkanoyl-CoA using:
- a polypeptide having thioesterase activity classified under EC 3.1.2.-;
- or
- a polypeptide having butanal dehydrogenase activity classified under EC 1.2.1.10 or EC 1.2.1.57 and a polypeptide having aldehyde dehydrogenase activity classified under EC 1.2.1.3 or EC 1.2.1.4.
A method of enzymatically producing 6-hydroxyhexanoate, the method comprising:
- a) enzymatically converting hexanoate to hexanoate methyl ester using a polypeptide having fatty acid O-methyltransferase activity classified under EC 2.1.1.15;
- b) enzymatically converting hexanoate methyl ester to 6-hydroxyhexanoate methyl ester using a polypeptide having monooxygenase activity classified under EC 1.14.14.- or EC 1.14.15.-;
- and
- c) enzymatically converting 6-hydroxyhexanoate methyl ester to 6-hydroxyhexanoate using a polypeptide having demethylase activity classified under EC 2.1.1.- or a polypeptide having esterase activity classified under EC 3.1.1.-.
Dependencies are inferred from the claim text.