Direct starch to fermentable sugar as feedstock for the production of isoprene, isoprenoid precursor molecules, and/or isoprenoids

A method for the production of isoprenoids by recombinant host cells in culture, wherein the host cells comprise one or more heterologous nucleic acids encoding one or more mevalonate (MVA) pathway polypeptides and one or more polyprenyl pyrophosphate synthase polypeptides, the method comprising:

• a. inactivating endogenous enzyme activity in a whole or fractionated grain;
• b. treating the whole or fractionated grain with a starch solubilizing alpha amylase and a glucoamylase, wherein the treatment is at a temperature below the gelatinization temperature of the starch in the grain, wherein the concentration of the alpha amylase is high, and wherein the treatment produces a fermentable sugar feedstock;
• c. culturing the recombinant host cells in culture media comprising the fermentable sugar feedstock;
• and
• d. producing isoprenoids.

A method for the production of mevalonate by recombinant host cells in culture, wherein the host cells comprise one or more heterologous nucleic acids encoding one or more mevalonate (MVA) pathway polypeptides, the method comprising:

• a. inactivating endogenous enzyme activity in a whole or fractionated grain;
• b. treating the whole or fractionated grain with a starch solubilizing alpha amylase and a glucoamylase, wherein the treatment is at a temperature below the gelatinization temperature of the starch in the grain, wherein the concentration of the alpha amylase is high, and wherein the treatment produces a fermentable sugar feedstock;
• c. culturing the recombinant host cells in culture media comprising the fermentable sugar feedstock;
• and
• d. producing mevalonate.