Microbial engineering for the production of isoprenoids
Disclosed herein are engineered cells and cell-free systems, compositions, and methods for conversion of isopentenols to isoprenoid precursors.
1. A host cell engineered to recombinantly express polynucleotides encoding Isopentenol Utilization Pathway (IUP) enzymes producing isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) from isoprenol and/or prenol, wherein the IUP enzymes comprise one or more heterologous enzymes, and the IUP enzymes comprise a Saccharomyces cerevisiae choline kinase, an isopentenyl phosphate kinase, and an isopentenyl pyrophosphate isomerase (IDI), and wherein when the host cell is Saccharomyces cerevisiae , the Saccharomyces cerevisiae host cell is transformed with an exogenous polynucleotide encoding the Saccharomyces cerevisiae choline kinase.
2. The host cell of claim 1 , wherein the S. cerevisiae choline kinase is encoded by a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 1.
3. The host cell of claim 1 , wherein the isopentenyl phosphate kinase is an enzyme of Haloferax volcanii, Methanothermobacter thermoautotrophicus, Methanocaldococcus janaschii, Arabidopsis thaliana , or Thermoplasma acidophilium.
4. The host cell of claim 3 , wherein the isopentenyl phosphate kinase is an enzyme of Arabidopsis thaliana.
5. The host cell of claim 1 , wherein the IDI is from Escherichia coli.
6. The host cell of claim 1 , wherein expression of the polynucleotides encoding the IUP enzymes is under the control of a constitutive promoter or a conditional promoter.
7. The host cell of claim 1 , wherein the polynucleotides encoding the IUP enzymes are inserted on a plasmid.
8. The host cell of claim 1 , wherein the polynucleotides encoding the IUP enzymes are integrated in the chromosome of the host cell.
9. The host cell of claim 1 , wherein the host cell is capable of producing the IPP and/or the DMAPP from isoprenol or prenol.
10. The host cell of claim 1 , wherein the host cell is a prokaryotic cell or a eukaryotic cell.
11. The host cell of claim 10 wherein the host cell is a fungal cell.
12. The host cell of claim 11 , wherein the fungal cell is a yeast cell selected from the group consisting of Saccharomyces spp., Schizosaccharomyces spp., Pichia spp., Paffia spp., Kluyveromyces spp., Candida spp., Talaromyces spp., Brettanornyces spp., Pachysolen spp., Debaryomyces spp., Yarrowia spp., and an industrial polyploid yeast strain.
13. The host cell of claim 1 wherein the host cell is Saccharomyces cerevisiae or Yarrowia lipolytica.
14. The host cell of claim 1 further comprising an isoprenoid synthesis pathway that converts the IPP and/or the DMAPP to an isoprenoid product.