Genetically modified host cells and use of same for producing isoprenoid compounds
AssigneeThe Regents of the University of California
InventorsJay D. Keasling, James Kirby, Eric M. Paradise
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Abstract
The present invention provides genetically modified eukaryotic host cells that produce isoprenoid precursors or isoprenoid compounds. A subject genetically modified host cell comprises increased activity levels of one or more of mevalonate pathway enzymes, increased levels of prenyltransferase activity, and decreased levels of squalene synthase activity. Methods are provided for the production of an isoprenoid compound or an isoprenoid precursor in a subject genetically modified eukaryotic host cell. The methods generally involve culturing a subject genetically modified host cell under conditions that promote production of high levels of an isoprenoid or isoprenoid precursor compound.
Claims22
An isolated eukaryotic host cell comprising one or more heterologous nucleic acids that are integrated into the host cell's chromosome wherein the chromosomally integrated heterologous nucleic acids encode:
- a) an enzyme that converts 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) to mevalonate;
- b) farnesyl diphosphate (FPP) synthase;
- and
- c) a repressible promoter that controls transcription of the endogenous gene encoding squalene synthase;
- wherein the host cell further comprises a heterologous terpene synthase,
- wherein the terpene synthase catalyzes conversion of FPP, produced by action of the FPP synthase, into an isoprenoid, and produces the isoprenoid under culture conditions that provide for repression of transcription of the endogenous squalene synthase-encoding gene,
- and wherein FPP does not accumulate at levels that are toxic to the cell.
A Saccharomyces cerevisiae cell comprising one or more heterologous nucleic acids that are integrated into the cell's chromosome wherein the chromosomally integrated heterologous nucleic acids encode:
- a) a truncated 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR);
- b) farnesyl diphosphate (FPP) synthase;
- and
- c) a repressible promoter that controls transcription of the endogenous gene encoding squalene synthase;
- wherein the host cell further comprises a heterologous terpene synthase,
- wherein the terpene synthase catalyzes conversion of FPP, produced by action of the FPP synthase, into an isoprenoid, and produces the isoprenoid under culture conditions that provide for repression of transcription of the endogenous squalene synthase-encoding gene,
- and wherein FPP does not accumulate at levels that are toxic to the cell.
Dependencies are inferred from the claim text.