Biosynthesis of isopentenyl pyrophosphate

A method for synthesizing an isoprenoid or an isoprenoid precursor via a mevalonate pathway in a host cell, wherein the method comprises:

• i) culturing a transformed host cell in a suitable medium, wherein the medium comprises DL-mevalonate, wherein the transformed host cell is a prokaryote that does not normally synthesize isopentenyl pyrophosphate (IPP) through the mevalonate pathway, and wherein the host cell comprises one or more nucleic acids heterologous to the host cell, wherein the one or more heterologous nucleic acids comprises nucleotide sequences that encode:
• (a) a eukaryotic enzyme that phosphorylates mevalonate to mevalonate 5-phosphate;
• (b) a eukaryotic enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate;
• and
• (c) a eukaryotic enzyme that converts mevalonate 5-pyrophosphate to IPP,
• wherein the host cell is not genetically modified with a heterologous nucleic acid encoding 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase (HMGS), HMG-CoA reductase, and acetoacetyl-CoA thiolase,
• said culturing providing for production of the enzymes, resulting in synthesis of said isoprenoid or isoprenoid precursor in a recoverable amount of at least about 1 mg/L;
• and
• ii) recovering the produced isoprenoid or isoprenoid precursor.

A method for synthesizing an isoprenoid or an isoprenoid precursor via a mevalonate pathway in a host cell, wherein the method comprises:

• i) culturing a transformed host cell in a suitable medium, wherein the medium comprises DL-mevalonate, wherein the transformed host cell is a prokaryote that does not normally synthesize isopentenyl pyrophosphate (IPP) through the mevalonate pathway, and wherein the host cell comprises one or more nucleic acids heterologous to the host cell, wherein the one or more heterologous nucleic acids comprises the nucleotide sequence set forth in SEQ ID NO:9 that comprises nucleotide sequences encoding:
• (a) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate;
• (b) an enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate;
• and
• (c) an enzyme that converts mevalonate 5-pyrophosphate to IPP,
• said culturing providing for production of the enzymes, resulting in synthesis of said isoprenoid or isoprenoid precursor in a recoverable amount of at least about 1 mg/L;
• and
• ii) recovering the produced isoprenoid or isoprenoid precursor.