(719f) Metabolic Engineering of Isoprenoid Biofuel Pathway

Fuels synthesis division at Joint BioEnergy Institute (JBEI) is responsible to develop and engineer a tool to convert processed biomass into biofuels. One potential biofuel targets are isoprenoid-based compounds, and mevalonate pathway has been explored and engineered as the major biosynthetic pathway for the production of isoprenoids fuel in both E. coli and yeast. The engineering of mevalonate pathway to produce more IPP (isopentenyl diphosphate) and DMAPP (3, 3-dimethylallyl diphosphate) is one of the main approach to improve isoprenoid based fuel production. In this study, we present several engineering strategies of top portion, which produces mevalonate from acetyl CoA, and of bottom portion of mevalonate pathway that produces IPP, DMAPP and FPP (farnesyl diphosphate) from mevalonate. Using proteomic analysis and synthetic biology tool, we addressed the major bottle necks of the pathway and engineered the proteins and genes to improve the production level. Also the combining entire mevalonate pathway genes in single plasmid using recently developed Biobrick standard cloning strategy has been performed, and the production of isoprenoids using this single plasmid versions of mevalonate pathway has been studied.