Production of mevalonate, isoprene, and isoprenoids using genes encoding polypeptides having thiolase, HMG-CoA synthase and HMG-CoA reductase enzymatic activities

Patent metadata

Field	Value
Publication number	US-10364443-B2
Application number	US-201715694193-A
Country	US
Kind code	B2
Filing date	Sep 1, 2017
Priority date	Apr 29, 2011
Publication date	Jul 30, 2019
Grant date	Jul 30, 2019

Abstract

The invention features compositions and methods for the increased production of mevalonate, isoprene, isoprenoid precursor molecules, and/or isoprenoids in microorganisms via the heterologous expression of the mvaE and mvaS genes from the organisms Listeria grayi DSM 20601, Enterococcus faecium, Enterococcus gallinarum EG2, and Enterococcus casseliflavus.

First claim

What is claimed is:

1. Recombinant cells with increased production of mevalonate, the cells comprising one or more heterologous nucleic acids comprising a mevalonate E (mvaE) gene and a mevalonate S (mvaS) gene selected from the organisms Listeria grayi ( L. grayi ), Enterococcus faecium ( E. faecium ), Enterococcus gallinarum ( E. gallinarum ), and Enterococcus casseliflavus ( E. casseliflavus ),
wherein the mvaE gene and mvaS gene encode polypeptides having thiolase, HMG-CoA synthase, and HMG-CoA reductase catalytic activities, and wherein the cells produce increased amounts of mevalonate compared to mevalonate-producing cells that (A) contain an mvaE gene and mvaS gene from Enterococcus faecalis ( E. faecalis ) and (B) do not contain said mvaE gene and mvaS gene from E. gallinarum, E. casseliflavus, E. faecium , or L. grayi.

2. The cells of claim 1 , wherein the one or more nucleic acids is placed under an inducible promoter or a constitutive promoter.

3. The cells of claim 1 , wherein the one or more nucleic acids is cloned into a multicopy plasmid.

4. The cells of claim 1 wherein the one or more nucleic acids is integrated into a chromosome of the cells.

5. The cells of claim 1 , wherein the cells are gram-positive bacterial cells, gram-negative bacterial cells, Escherichia cells, Pantoea cells, fungal cells, filamentous fungal cells, Trichoderma cells, Aspergillus cells, or yeast cells.

6. The cells of claim 5 , 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.

7. The cells of claim 6 , wherein the cells are E. coli.

8. The cells of claim 1 , wherein the mvaE gene and mvaS gene are from different organisms.

9. The cells of claim 1 , wherein
a) the mvaE gene is from L. grayi and the mvaS gene is from E. faecium;
b) the mvaE gene is from L. grayi and the mvaS gene is from E. gallinarum;
c) the mvaE gene is from L. grayi and the mvaS gene is from E. casseliflavus;
d) the mvaE gene is from E. faecium and the mvaS gene is from L. grayi;
e) the mvaE gene is from E. faecium and the mvaS gene is from E. gallinarum;
f) the mvaE gene is from E. faecium and the mvaS gene is from E. casseliflavus;
g) the mvaE gene is from E. gallinarum and the mvaS gene is from L. grayi;
h) the mvaE gene is from E. gallinarum and the mvaS gene is from E. faecium;
i) the mvaE gene is from E. gallinarum and the mvaS gene is from E. casseliflavus;
j) the mvaE gene is from E. casseliflavus and the mvaS gene is from L. grayi;
k) the mvaE gene is from E. casseliflavus and the mvaS gene is from E. faecium ; or
l) the mvaE gene is from E. casseliflavus and the mvaS gene is from E. gallinarum.

10. The cells of claim 5 , wherein the cells are yeast cells.

11. The cells of claim 10 , wherein the yeast cells are selected from the group consisting of Saccharomyces sp., Schizosaccharomyces sp., Pichia sp., and Candida sp. cells.

12. The cells of claim 11 , wherein the yeast cells are Saccharomyces cerevisiae cells.

13. A method of producing mevalonate, comprising: (a) culturing the host cells of claim 1 under suitable culture conditions for production of mevalonate; and (b) producing the mevalonate.

14. The method of claim 13 , further comprising (c) recovering the mevalonate.

Assignees

• Danisco Us Inc
• Goodyear Tire & Rubber

Inventors

• Beck Zachary Q
• Miller Michael C
• Peres Caroline M
• Primak Yuliya A
• Pucci Jeff P
• Wells Derek H

Classifications

• C12Y203/01016

  Acetyl-CoA C-acyltransferase (2.3.1.16) · CPC title

• C12P7/02

  containing a hydroxy group · CPC title

• C12N15/81

  for yeasts · CPC title

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title