Method for enhancing production of isoprenoid compounds

A method for synthesizing isopentenyl pyrophosphate (IPP) via a mevalonate pathway in a host cell, the method comprising culturing a genetically modified host cell in a suitable medium, wherein the genetically modified host cell produces IPP via a mevalonate pathway, wherein the genetically modified host cell comprises one or more mutations in an endogenous nucleic acid that comprises a nucleotide sequence encoding encoding one or more enzymes that, when produced in the cell, reduce HMG-CoA accumulation-induced growth inhibition, compared to a control parent host cell that is not genetically modified to comprise the one or more mutations and that exhibits HMG-CoA accumulation-induced growth inhibition, and wherein the level of IPP produced in the genetically modified host cell is higher than the level of IPP produced in the control parent host cell.

A method for enhancing production of an isoprenoid or isoprenoid precursor via a mevalonate pathway in a host cell, the method comprising culturing a genetically modified host cell in a suitable medium, wherein the genetically modified host cell produces the isoprenoid or isoprenoid precursor via a mevalonate pathway, wherein the genetically modified host cell comprises one or more heterologous nucleic acids comprising nucleotide sequences encoding one or more enzymes that, when produced in the cell, reduce HMG-CoA accumulation-induced growth inhibition, compared to a control parent host cell that is not genetically modified with the one or more heterologous nucleic acids and that exhibits HMG-CoA accumulation-induced growth inhibition, and wherein the level of isoprenoid or isoprenoid precursor produced in the genetically modified host cell is higher than the level of isoprenoid or isoprenoid precursor produced in the control parent host cell.

A method of identifying a nucleic acid comprising a nucleotide sequence encoding a variant HMG-CoA reductase (HMGR) that reduces intracellular HMG-CoA accumulation, the method comprising:

• a) producing a test cell by introducing into a host cell an exogenous nucleic acid comprising a nucleotide sequence encoding a candidate gene product, wherein the host cell produces mevalonate via a mevalonate pathway and exhibits HMG-CoA accumulation-induced growth inhibition;
• and
• b) determining the effect, if any, of expression of the candidate gene product on growth of the test cell, wherein a reduction in growth inhibition indicates the candidate gene product encodes a variant HMGR that reduces intracellular HMG-CoA accumulation.

A method of identifying an agent that reduces the HMG-CoA accumulation in a cell, the method comprising:

• a) contacting a test cell with a test agent, wherein the test cell synthesizes mevalonate via a mevalonate pathway, and wherein the test cell exhibits HMG-CoA accumulation-induced growth inhibition;
• and
• b) determining the effect, if any, of the test agent on HMG-CoA accumulation-induced growth inhibition, wherein a reduction in growth inhibition indicates that the candidate agent reduces the levels of HMG-CoA accumulation in the cell.