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US 10,167,487 B2Granted
Methods, cells and reagents for production of isoprene, derivatives and intermediates thereof
AssigneeInvista North America S.A.R.L.
InventorAlex Van Eck Conradie
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Abstract
This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy-3-methylglutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methylglutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene.
Claims17
A method for synthesizing 3-hydroxy-3-methylglutaryl-CoA comprising:
- enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbut-2-enoyl-CoA by:
- (a) enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbutanal;
- enzymatically converting 3-methylbutanal to 3-methylbutanoate;
- and enzymatically converting 3-methylbutanoate to 3-methylbutanoyl-CoA;
- (b) enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbutanoyl-CoA;
- or
- (c) both (a) and (b);
- enzymatically converting 3-methylbutanoyl-CoA to 3-methylbut-2-enoyl-CoA;
- enzymatically converting 3-methylbut-2-enoyl-CoA to 3-methyl-glutaconyl CoA;
- and
- enzymatically converting 3-methyl-glutaconyl-CoA to 3-hydroxy-3-methylglutaryl-CoA.
8 more elements
A method for synthesizing 3-hydroxy-3-methylglutaryl-CoA comprising:
- enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbut-2-enoyl-CoA by:
- (a) enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbutanal using a polypeptide having the activity of an EC 4.1.1.74 or EC 4.1.1.43 enzyme;
- enzymatically converting 3-methylbutanal to 3-methylbutanoate using a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme;
- and
- enzymatically converting 3-methylbutanoate to 3-methylbutanoyl-CoA using a polypeptide having the activity of an EC 6.2.1.2 enzyme;
- (b) enzymatically converting 4-methyl-2-oxopentanoate to 3-methylbutanoyl-CoA using a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1 enzyme;
- or
- (c) both (a) and (b);
- enzymatically converting 3-methylbutanoyl-CoA to 3-methylbut-2-enoyl-CoA using a polypeptide having the activity of an EC 1.3.8.4 enzyme;
- enzymatically converting 3-methylbut-2-enoyl-CoA to 3-methyl-glutaconyl CoA using a polypeptide having the activity of an EC 6.4.1.4 enzyme;
- and
- enzymatically converting 3-methyl-glutaconyl-CoA to 3-hydroxy-3-methylglutaryl-CoA using a polypeptide having enzyme activity classified under EC 4.2.1.18.
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Dependencies are inferred from the claim text.