Microorganisms and processes for the production of isoprene

1. A non-naturally occurring microbial organism comprising an isoprene biosynthetic pathway for conversion of dimethylallyl diphosphate (DMAPP) to isoprene, wherein the isoprene biosynthetic pathway comprises the conversion of DMAPP to 3-methyl-2-buten-1-ol by a 3-methyl-2-buten-1-ol synthase, followed by the conversion of 3-methyl-2-buten-1-ol to 2-methyl-3-buten-2-ol by a 2-methyl-3-buten-2-ol isomerase, followed by the conversion of 2-methyl-3-buten-2-ol to isoprene by a 2-methyl-3-buten-2-ol dehydratase, wherein the organism comprises an exogenous nucleic acid encoding a 3-methyl-2-buten-1-ol synthase; a 2-methyl-3-buten-2-ol isomerase; and a 2-methyl-3-buten-2-ol dehydratase; and wherein the isoprene biosynthetic pathway is expressed at a sufficient level to produce isoprene.

2. The non-naturally occurring microbial organism of claim 1 , wherein the organism overexpresses one or more endogenous or exogenous genes encoding at least one enzyme selected from: an enzyme of the methylerythritol phosphate pathway or an enzyme of the mevalonate pathway.

3. The non-naturally occurring microbial organism of claim 2 , wherein the dimethylallyl diphosphate available for conversion to isoprene is increased.

4. The non-naturally occurring microbial organism of claim 1 , wherein the 2-methyl-3-buten-2-ol dehydratase is a bi-functional enzyme further comprising 2-methyl-3-buten-2-ol isomerase activity.

5. The non-naturally occurring microbial organism of claim 4 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase.

6. The non-naturally occurring microbial organism of claim 5 , wherein the 2-methyl-3-buten-2-ol dehydratase is a linalool dehydratase-isomerase derived from Castellaniella defragrans.

7. The non-naturally occurring microbial organism of claim 1 , wherein the 3-methyl-2-buten-1-ol synthase is a phosphatase.

8. The non-naturally occurring microbial organism of claim 7 , wherein the phosphatase is derived from Bacillus subtilis yqkG, Bacillus subtilis yhfR, or Escherichia coli ytjC.

9. The non-naturally occurring microbial organism of claim 1 , wherein the 3-methyl-2-buten-1-ol synthase is a terpene synthase.

10. The non-naturally occurring microbial organism of claim 9 , wherein the terpene synthase is a geraniol synthase.

11. The non-naturally occurring microbial organism of claim 10 , wherein the geraniol synthase is derived from Ocimum basilicum, Perilla citriodora, Perilla frutescans , or Cinnamomom tenuipile.

12. The non-naturally occurring microbial organism of claim 9 , wherein the terpene synthase is a farnesol synthase.

13. The non-naturally occurring microbial organism of claim 12 , wherein the farnesol synthase is derived from Zea mays or Oryza satiza.

14. The non-naturally occurring microbial organism of claim 1 , further comprising one or more endogenous or exogenous genes encoding at least one enzyme of the methylerythritol phosphate pathway selected from: 1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose-5-phosphate reductoisomerase, 4-diphosphocytidyl-2-C-methyl-D-erythritol synthase, 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase, 2-C-methyl-D-erythritol-2,4-cyclodiphosphate synthase, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase, dimethylallyl-diphosphate/isopentenyl-diphosphate:NAD(P) + oxidoreductase, isopentenyl diphosphate isomerase, and combinations thereof.

15. The non-naturally occurring microbial organism of claim 1 , further comprising one or more endogenous or exogenous genes encoding at least one enzyme of the mevalonate pathway selected from: acetyl-CoA acetyltransferase, 3-hydroxy-3-methylglutaryl-CoA synthase, 3-hydroxy-3-methylglutaryl-CoA reductase, mevalonate kinase, phosphomevalonate kinase, diphosphomevalonate decarboxylase, isopentenyl-diphosphate isomerase and combinations thereof.

16. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism is selected from bacteria, archaea, eubacteria, yeast and fungi.

17. The non-naturally occurring microbial organism of claim 1 , wherein the microbial organism is an Escherichia coli.

18. The non-naturally occurring microbial organism of claim 2 , wherein the at least one enzyme of the methylerythritol phosphate pathway is selected from: 1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose-5-phosphate reductoisomerase, 4-diphosphocytidyl-2-C-methyl-D-erythritol synthase, 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase, 2-C-methyl-D-erythrito 1 -2,4-cyclodiphosphate synthase, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase, dimethylallyl-diphosphate/isopentenyl-diphosphate:NAD(P) + oxidoreductase, isopentenyl diphosphate isomerase, and combinations thereof.

19. The non-naturally occurring microbial organism of claim 18 , wherein the at least one enzyme of the methylerythritol phosphate pathway is selected from: 1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose-5-phosphate reductoisomerase and isopentenyl diphosphate isomerase.

20. The non-naturally occurring microbial organism of claim 2 , wherein the at least one enzyme of the mevalonate pathway is selected from: acetyl-CoA acetyltransferase, 3-hydroxy-3-methylglutaryl-CoA synthase, 3-hydroxy-3-methylglutaryl-CoA reductase, mevalonate kinase, phosphomevalonate kinase, diphosphomevalonate decarboxylase, isopentenyl-diphosphate isomerase and combinations thereof.

21. A non-naturally occurring microbial organism of claim 1 , wherein the dimethylallyl diphosphate available for conversion to isoprene is increased.

22. A method of producing isoprene, the method comprising the steps of culturing a non-naturally occurring microbial organism of claim 1 in a suitable culture medium containing a carbon source under conditions such that the non-naturally occurring microorganism converts at least a part of the carbon source to isoprene, and optionally recovering the isoprene.

23. The method of claim 22 , wherein at least one enzyme of a methylerythritol phosphate pathway or the mevalonate pathway is overexpressed by the organism.