Methods of producing 7-carbon chemicals via CoA-dependent carbon chain elongation associated with carbon storage
AssigneeInvista North America S.A.R.L.
InventorsAlex Van Eck Conradie, Adriana Leonora Botes
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Abstract
This document describes biochemical pathways for producing pimelic acid, 7-hydroxyheptanoic acid, 7-aminoheptanoic acid, heptamethylenediamine or 1,7-heptanediol by forming two terminal functional groups, comprised of carboxyl, amine or hydroxyl group, in a C7 aliphatic backbone substrate. These pathways, metabolic engineering and cultivation strategies described herein rely on the CoA-dependent elongation enzymes or analog enzymes associated with the carbon storage pathways from polyhydroxyalkanoate accumulating bacteria.
Claims22
A method for biosynthesizing 7-aminoheptanoate in vitro or in a recombinant host, said method comprising:
- providing acetyl-CoA and propanoyl-CoA;
- enzymatically synthesizing heptanoyl-CoA from acetyl-CoA and propanoyl-CoA via two cycles of CoA-dependent carbon chain elongation;
- enzymatically converting heptanoyl-CoA to heptanoate by contacting heptanoyl-CoA with a butanal dehydrogenase classified under EC 1.2.1.57 to form heptanal and contacting heptanal with an aldehyde dehydrogenase classified under EC 1.2.1.4 to form heptanoate, or contacting heptanoyl-CoA with a thioesterase classified under EC 3.1.2.- to form heptanoate;
- enzymatically converting heptanoate to 7-hydroxyheptanoate by contacting heptanoate with a cytochrome P450 monooxygenase classified under EC 1.14.15.1 or EC 1.14.15.3 to form 7-hydroxyheptanoate;
- enzymatically converting 7-hydroxyheptanoate to pimelate semialdehyde by contacting 7-hydroxyheptanoate with an alcohol dehydrogenase classified under EC 1.1.1.- or a cytochrome P450 monooxygenase classified under EC 1.14.15.1 or EC 1.14,15.3;
- and
- enzymatically converting pimelate semialdehyde to 7-aminoheptanoate by contacting pimelate semialdehyde with a ω-transaminase classified under EC 2.6.1.-.
A recombinant host comprising one or more nucleic acids encoding each of the following enzymes:
- (i) a β-ketothiolase classified under EC 2.3.1.16;
- or a β-ketoacyl-[acp] synthase classified under EC 2.3.1.180 and an acetyl-CoA carboxylase classified under EC 6.4.1.2;
- (ii) a 3-hydroxyacyl-CoA dehydrogenase classified under EC 1.1.1.157, EC 1.1.1.36, or EC 1.1.1.35;
- or a 3-oxoacyl-CoA reductase classified under EC 1.1.1.100;
- (iii) an enoyl-CoA hydratase classified under EC 4.2.1.17 or EC 4.2.1.119;
- and (iv) a trans-2-enoyl-CoA reductase classified under EC 1.3.1.8, EC 1.3.1.38, or EC 1.3.1.44, wherein at least one enzyme is encoded by an exogenous nucleic acid, said host producing heptanoyl-CoA.
Dependencies are inferred from the claim text.