Compositions and methods for producing isoprene

Patent metadata

Field	Value
Publication number	US-2016281113-A1
Application number	US-201614987681-A
Country	US
Kind code	A1
Filing date	Jan 4, 2016
Priority date	Dec 13, 2007
Publication date	Sep 29, 2016
Grant date	—

Abstract

The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells.

First claim

1 - 20 . (canceled)

21 . Recombinant cells capable of producing isoprene, the cells comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide, wherein the cells (i) produce greater than about 400 nmole/g wcm /hr of isoprene, (ii) convert more than about 0.002 molar percent of the carbon that the cells consume from a cell culture medium into isoprene, or (iii) have an average volumetric productivity of isoprene greater than about 0.1 mg/L broth /hr.

22 . The cells of claim 21 , wherein the heterologous nucleic acid encoding an isoprene synthase polypeptide is operably linked to a promoter.

23 . The cells of claim 21 , wherein the isoprene synthase polypeptide is a plant isoprene synthase polypeptide.

24 . The cells of claim 23 , wherein the plant isoprene synthase polypeptide is a poplar isoprene synthase polypeptide or a kudzu isoprene synthase polypeptide.

25 . The cells of claim 21 , wherein the heterologous nucleic acid encoding an isoprene synthase polypeptide is in a vector or is integrated into a chromosome of the cells.

26 . The cells of claim 21 , further comprising nucleic acids encoding: (a) an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide or (b) at least one of a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide and/or one or more mevalonate (MVA) pathway polypeptides.

27 . The cells of claim 26 , wherein at least one of the nucleic acids encoding one or more MVA pathway polypeptides of (b) is a heterologous nucleic acid or a copy of an endogenous nucleic acid.

28 . The cells of claim 26 , wherein the cells comprise polypeptides of the entire MVA pathway.

29 . The cells of claim 26 , wherein at least one of the nucleic acids encoding a polypeptide of (a) and (b) is in a vector or is integrated into a chromosome of the cells.

30 . The cells of claim 26 , wherein at least one of the nucleic acids encoding a polypeptide of (a) and (b) is over-expressed.

31 . The cells of claim 30 , wherein the over-expressed nucleic acid is cloned into a multicopy plasmid.

32 . The cells of claim 30 , wherein the over-expressed nucleic acid is placed under an inducible promoter or a constitutive promoter.

33 . The cells of claim 21 , wherein the cells are gram-positive bacterial cells or gram-negative bacterial cells.

34 . The cells of claim 21 , wherein the 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., and P. alcaligenes cells.

35 . The cells of claim 21 , wherein the cells are fungal cells.

36 . The cells of claim 35 , wherein the fungal cells are Aspergillus , yeast, Trichoderma , or Yarrowia cells.

37 . The cells of claim 36 , where the yeast cells are Saccharomyces sp., Schizosaccharomyces sp., Pichia sp., or Candida sp. cells.

38 . The cells of claim 35 , where the fungal 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. oryzae, A. niger, 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.

39 . The cells of claim 21 , wherein the cells are algal cells.

40 . The cells of claim 39 , wherein the algal cells are selected from the group consisting of: green algae, red algae, glaucophytes, chlorarachniophytes, euglenids, chromista, and dinoflagellates.

41 . A method for producing isoprene, the method comprising (a) culturing the recombinant cells of claim 21 under suitable conditions for the production of isoprene; and (b) producing isoprene.

42 . The method of claim 41 , further comprising (c) recovering the isoprene.

Assignees

• Danisco Us Inc
• Goodyear Tire & Rubber

Inventors

• Cervin Marguerite A
• Chotani Gopal K
• Feher Frank J
• La Duca Richard
• Mcauliffe Joseph C
• Miasnikov Andrei
• Peres Caroline M
• Puhala Aaron S
• Sanford Karl J
• Valle Fernando
• Whited Gregory M

Classifications

• C12N9/88

  Lyases (4.) · CPC title

• C12Y203/01009

  Acetyl-CoA C-acetyltransferase (2.3.1.9) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12Y207/01036

  Mevalonate kinase (2.7.1.36) · CPC title

• C12P5/007 Primary

  containing one or more isoprene units, i.e. terpenes (carotenes C12P23/00) · CPC title