Engineering of the pyruvate dehydrogenase bypass in Saccharomyces cerevisiae for high-level production of isoprenoids.
Amorphadiene, a sesquiterpene precursor to the anti-malarial drug artemisinin, is synthesized by the cyclization of farnesyl pyrophosphate (FPP). Saccharomyces cerevisiae produces FPP through the mevalonate pathway using acetyl-CoA as a starting compound. In order to enhance the supply of acetyl-CoA to the mevalonate pathway and achieve high-level production of amorphadiene, we engineered the pyruvate dehydrogenase bypass in S. cerevisiae. Overproduction of acetaldehyde dehydrogenase and introduction of a Salmonella enterica acetyl-CoA synthetase variant increased the carbon flux into the mevalonate pathway resulting in increased amorphadiene production. This work will be generally applicable to the production of a broad range of isoprenoids in yeast.
Shiba
Yoichiro
Y
Berkeley Center for Synthetic Biology, Department of Chemical Engineering, University of California, 717 Potter Street, Building 977, Mail code 3224, Berkeley, CA 94720-3224, USA.
Paradise
Eric M
EM
Kirby
James
J
Ro
Dae-Kyun
DK
Keasling
Jay D
JD
D000105:Acetyl Coenzyme A / Q000235:genetics*; D000445:Aldehyde Oxidoreductases / Q000235:genetics*; D024861:Genetic Enhancement / Q000379:methods*; D015202:Protein Engineering / Q000379:methods*; D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D013729:Terpenes / Q000378:metabolism*
D011768:Pyruvate Dehydrogenase Complex; D011994:Recombinant Proteins; D013729:Terpenes; D000105:Acetyl Coenzyme A; D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+)