The goal of this project is to develop a host strain and expression system that will allow high-level production of natural and un-natural terpenoids in Escherichia coli. The specific aims of this project are to: (1) maximize the production of the isoprenoid precursor isopentyl diphosphate in E. coli by expressing the genes for either the mevalonate-dependent or the mevalonate-independent synthesis pathway using the metabolic engineering tools developed in the Principal Investigator's laboratory; (2) maximize production of the primary precursors to the terpenoids: geranyl diphosphate, farnesyl diphosphate, and geranylgernyldiphosphate; (3) introduce into E. coli the genes for specific classes of terpenoids and optimize production of these "natural" terpenoids; and (4) use laboratory evolution of terpene cyclases to produce novel terpenoids or to change the distribution of products made by terpenoid biosynthetic enzymes. This project is part of an Interagency Activity in Metabolic Engineering and is to be funded by the Office of Naval Research (ONR) and three Programs within NSF.
