Production of mevalonate, isoprene, and isoprenoids using genes encoding polypeptides having thiolase, hmg-coa synthase and hmg-coa reductase enzymatic activities
AssigneesDanisco US Inc., The Goodyear Tire & Rubber Company
InventorsZachary Q. BECK, Michael C. MILLER, Caroline M. PERES, Yuliya A. PRIMAK, Jeff P. PUCCI, Derek H. WELLS
Life of this patent
Estimating the term…
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.
Claims45
Recombinant cells capable of increased production of isoprene, the cells comprising one or more heterologous nucleic acids comprising nucleotide sequences selected from the group consisting of:
- a. a mvaE gene and a mvaS gene from E. gallinarum;
- b. a mvaE gene and a mvaS gene from E. casseliflavus;
- c. a mvaE gene and a mvaS gene from E. faecium;
- and
- d. a mvaE gene and a mvaS gene from L. grayi
- wherein said mvaE gene and mvaS gene encode polypeptides having thiolase, HMG-CoA synthase, and HMG-CoA reductase catalytic activities, and wherein the cells further comprise:
- i. one or more nucleic acids encoding polypeptides of the lower MVA pathway;
- and
- ii. a heterologous nucleic acid encoding an isoprene synthase polypeptide,
- wherein the cells produce increased amounts of isoprene compared to isoprene-producing cells that do not comprise said mvaE gene and mvaS gene.
Recombinant cells capable of increased production of isoprenoid precursors, the cells comprising one or more heterologous nucleic acids comprising nucleotide sequences selected from the group consisting of:
- (a) an mvaE gene and an mvaS gene from E. gallinarum;
- (b) an mvaE gene and an mvaS gene from E. casseliflavus;
- (c) an mvaE gene and an mvaS gene from E. faecium;
- and
- (d) an mvaE gene and an mvaS gene from L. grayi,
- 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 increase amounts of isoprenoid precursors compared to isoprenoid precursor-producing cells that do not comprise said mvaE gene and mvaS gene.
Recombinant cells capable of increased production of isoprenoids, the cells comprising one or more heterologous nucleic acids comprising nucleotide sequences selected from the group consisting of:
- (a) an mvaE gene and an mvaS gene from E. gallinarum;
- (b) an mvaE gene and an mvaS gene from E. casseliflavus;
- (c) an mvaE gene and an mvaS gene from E. faecium;
- and
- (d) an mvaE gene and an mvaS gene from L. grayi,
- wherein said mvaE gene and mvaS gene encode polypeptides having thiolase, HMG-CoA synthase, and HMG-CoA reductase catalytic activities, and wherein the cell further comprise:
- i. one or more nucleic acids encoding polypeptides of the lower MVA pathway;
- and
- ii. one or more nucleic acids encoding polyprenyl pyrophosphate synthases,
- wherein the cells produce increased amounts of isoprenoids compared to isoprenoid-producing cells that do not comprise said mvaE gene and mvaS gene.
Dependencies are inferred from the claim text.