Compositions and methods for producing isoprene
The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells.
1. A method of producing cis-polyisoprene, the method comprising:
(a) culturing recombinant cells under suitable culture conditions for the production of isoprene, wherein the cells comprise nucleic acids encoding
(i) a heterologous isoprene synthase polypeptide; and
(ii) two or more heterologous mevalonate (MVA) pathway polypeptides, and
wherein the cells (1) produce greater than 400 nmole/gwcm/hr of isoprene, (2) convert more than 0.002 molar percent of the carbon that the cells consume from a cell culture medium into isoprene, (3) have an average volumetric productivity of isoprene greater than 0.1 mg/Lbroth/hr of isoprene, or (4) produce at least 0.0037 grams of isoprene per gram of dry cell mass (gdcm);
(b) producing isoprene; and
(c) polymerizing the isoprene to produce cis-polyisoprene.
2. The method of claim 1 , wherein the cells further comprise an endogenous nucleic acid encoding an isoprene synthase polypeptide.
3. The method of claim 1 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.
4. The method of claim 3 , wherein the plant isoprene synthase polypeptide is a poplar isoprene synthase polypeptide or a kudzu isoprene synthase polypeptide.
5. The method of claim 1 , wherein the cells further comprise nucleic acids encoding: (a) an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide or (b) a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide.
6. The method of claim 1 , wherein the cells comprise polypeptides of the entire MVA pathway.
7. The method of claim 1 , wherein the cells are fungal, bacterial, plant, or algal cells.
8. The method of claim 7 , wherein the cells are fungal cells.
9. The method of claim 8 , wherein the cells are Aspergillus , yeast, or Trichoderma cells.
10. The method of claim 9 , wherein the yeast cells are Saccharomyces sp., Schizosaccharomyces sp., Pichia sp., Yarrowia sp., or Candida sp. cells.
11. The method of claim 8 , where the cells are selected from the group consisting of A. oryzae, A. niger, S. cerevisiae, S. pombe, T. reesei, H. insolens, H. lanuginose, H. grisea, C. lucknowense, A. sojae, A. japonicus, A. nidulans, A. aculeatus, A. awamori, F. roseum, F. graminum, F. cerealis, F. oxysporuim, F. venenatum, N. crassa, M. miehei, T. viride, F. oxysporum , and F. solan cells.
12. The method of claim 7 , wherein the cells are bacterial cells.
13. The method of claim 12 , wherein the cells are gram-positive bacterial cells or gram-negative bacterial cells.
14. The method of claim 13 , wherein the bacterial cells are selected from the group consisting of E. coli, P. citrea, B. subtilis, B. licheniformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. clausii, B. halodurans, B. megaterium, B. coagulans, B. circulans, B. lautus, B. thuringiensis, S. albus, S. lividans, S. coelicolor, S. griseus, Pseudomonas sp., Corynebacteria sp., and P. alcaligenes cells.
15. The method of claim 7 , wherein the cells are plant cells.
16. The method of claim 15 , wherein the plant cells are selected from the group consisting of: the family Fabaceae, the Faboideae subfamily, kudzu, poplar, Populus alba x tremula, Populus alba , aspen, Populus tremuloides , and Quercus robur cells.
17. The method of claim 7 , wherein the cells are algal cells.
18. The method of claim 17 , wherein the algal cells are selected from the group consisting of: green algae, red algae, glaucophytes, chlorarachniophytes, euglenids, chromista, and dinoflagellates.
19. The method of claim 1 , further comprising (d) producing a tire from the cis-polyisoprene.