Method for enhancing production of isoprenoid compounds
Patent number: 7183089 Filing date: May 20, 2005
Issue date: Feb 27, 2007
Inventors: Jay D. Keasling, Jack D. Newman, Douglas J. Pitera
Assignee: The Regents of the University of California
A method for producing an isoprenoid or isoprenoid precursor via a mevalonate pathway in Escherichia coli, the method comprising:
(i) culturing in a suitable medium an E. coli cell genetically modified to produce heterologous hydroxymethylglutaryl-CoA (HMG-CoA) synthase (HMGS) and heterologous HMG-CoA reductase (HMGR), wherein the relative activity level of the HMGR produced in the genetically modified E. coli cell is higher than the relative activity level of the HMGS produced in the genetically modified E. coli cell, and wherein the genetically modified E. coli cell is further genetically modified to produce one or more additional heterologous mevalonate pathway enzymes selected from:
(a) an acetoacetyl-CoA thiolase;
(b) a mevalonate kinase;
(c) a phosphomevalonate kinase; and
(d) a mevalonate pyrophosphate decarboxylase;
wherein the genetically modified E. coli cell produces the isoprenoid or isoprenoid precursor at a level at least as high as the level produced in a control E. coli strain expressing pBAD33MevT sequence shown in SEQ ID NO:2, and wherein the genetically modified E. coli cell exhibits less HMG-CoA induced growth inhibition compared to the control E. coli strain; and
(ii) recovering the produced isoprenoid or isoprenoid precursor.
Patent number: 7183089 Filing date: May 20, 2005
Issue date: Feb 27, 2007
Inventors: Jay D. Keasling, Jack D. Newman, Douglas J. Pitera
Assignee: The Regents of the University of California
A method for producing an isoprenoid or isoprenoid precursor via a mevalonate pathway in Escherichia coli, the method comprising:
(i) culturing in a suitable medium an E. coli cell genetically modified to produce heterologous hydroxymethylglutaryl-CoA (HMG-CoA) synthase (HMGS) and heterologous HMG-CoA reductase (HMGR), wherein the relative activity level of the HMGR produced in the genetically modified E. coli cell is higher than the relative activity level of the HMGS produced in the genetically modified E. coli cell, and wherein the genetically modified E. coli cell is further genetically modified to produce one or more additional heterologous mevalonate pathway enzymes selected from:
(a) an acetoacetyl-CoA thiolase;
(b) a mevalonate kinase;
(c) a phosphomevalonate kinase; and
(d) a mevalonate pyrophosphate decarboxylase;
wherein the genetically modified E. coli cell produces the isoprenoid or isoprenoid precursor at a level at least as high as the level produced in a control E. coli strain expressing pBAD33MevT sequence shown in SEQ ID NO:2, and wherein the genetically modified E. coli cell exhibits less HMG-CoA induced growth inhibition compared to the control E. coli strain; and
(ii) recovering the produced isoprenoid or isoprenoid precursor.
