The invention relates, in part, to
nucleic acid constructs, genetically modified host cells and methods employing such constructs and host cells to increase the production of 3-methyl-2-butenol from IPP. Thus, in some aspects, the invention provides a genetically modified host
cell transformed with a
nucleic acid construct encoding a
fusion protein comprising a
phosphatase capable of catalyzing the
dephosphorylation of dimethylallyl diphosphate (DMAPP) linked to an IPP
isomerase capable of converting IPP to DMAPP, wherein the
nucleic acid construct is operably linked to a
promoter. In some embodiments, the genetically modified host
cell 5 further comprises a nucleic acid encoding a
reductase that is capable of converting 3-methyl-2-butenol to 3-methyl-
butanol. In some embodiments, the
reductase is encoded by a nucleic acid construct introduced into the
cell. In some embodiments, the IPP
isomerase is a Type I
isomerase. In some embodiments, the IPP isomerase is a Type II isomerase. In some embodiments, the host cell is selected from a group of taxonimcal classes consisting of 20
Escherichia,
Enterobacter,
Azotobacter,
Erwinia, Bacillus,
Pseudomonas,
Klebsiella,
Proteus,
Salmonella,
Serratia,
Shigella,
Rhizobia,
Vitreoscilla,
Synechococcus,
Synechocystis, and
Paracoccus taxonomical classes. In some embodiments, the host cell is an
Escherichia coli cell. In some embodiments, the host cell is a fungal cell, such as a
yeast cell. In some embodiments, the
yeast cell is a
Saccharomyces sp. cell. In some embodiments, the host cell is an algal,
insect or
mammalian cell line. In some embodiments, the
phosphatase is nudB from E. coli. In some embodiments, the IPP isomerase is encoded by an idi
gene from E. coli or idil
gene from
Saccharomyces cerevisiae.