Production of isoprenoids

摘要

Methods for producing an isoprenoid are provided. A plurality of bacterial or fungal host cells is obtained. These cells comprise a heterologous nucleic acid encoding one or more enzymes of a mevalonate pathway for making isopentenyl pyrophosphate. Expression of the one or more enzymes is under control of at least one heterologous transcriptional regulator. The mevalonate pathway comprises (i) an enzyme that condenses acetoacetyl-CoA with acetyl-CoA to form HMG-CoA, (ii) an enzyme that converts HMG-CoA to mevalonate, (iii) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate, (iv) an enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate, and (v) an enzyme that converts mevalonate 5-pyrophosphate to isopentenyl pyrophosphate. The host cells are cultured in a medium under conditions that are suboptimal as compared to conditions for the maximum growth rate. Temperature is maintained at a level below that which would provide for a maximum specific growth rate for the host cells.

主权项

1. A method for producing an isoprenoid comprising:
(a) obtaining a plurality of bacterial or fungal host cells that comprise a heterologous nucleic acid encoding one or more enzymes of a mevalonate pathway for making isopentenyl pyrophosphate, wherein expression of the one or more enzymes is under control of at least one heterologous transcriptional regulator, and wherein said mevalonate pathway comprises (i) an enzyme that condenses acetoacetyl-CoA with acetyl-CoA to form HMG-CoA; (ii) an enzyme that converts HMG-CoA to mevalonate; (iii) an enzyme that phosphorylates mevalonate to mevalonate 5-phosphate; (iv) an enzyme that converts mevalonate 5-phosphate to mevalonate 5-pyrophosphate; and (v) an enzyme that converts mevalonate 5-pyrophosphate to isopentenyl pyrophosphate; and (b) culturing the host cells in a medium wherein temperature is maintained at a level which would provide for about 90% or less of a maximum specific growth rate for the plurality of bacterial or fungal host cells.