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.
Claims35
1-4. (canceled)
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.
6-8. (canceled)
10-12. (canceled)
A non-naturally occurring host capable of producing 3-hydroxy-3-methylglutaryl-CoA, said host comprising:
- (a) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.-enzyme;
- or
- (b) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1,74 or EC 4.1.1.43 enzyme;
- at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme;
- at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 8,2.1.2. enzyme;
- or
- (c) both (a) and (b);
- and
- at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.3.8.4 enzyme;
- at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 8.4.1.4 enzyme;
- and
- at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.2.1.18 enzyme.
(canceled)
(canceled)
Dependencies are inferred from the claim text.