Synthetic Biology of Yeast

The goal of the proposed work is to develop a suite of tools for engineering the metabolism of Saccharomyces cerevisiae and demonstrate their use in expressing hybrid PKSs to produce a variety of mono- and diacids. The specific aims of the proposed work are as follows. 1) We will develop generalized methods for rapidly modifying the S. cerevisiae genome and demonstrate their use in developing a chassis capable of growing and/or surviving at low pH. 2) We will develop a suite of gene expression control devices for S. cerevisiae and demonstrate their utility for producing large, multicomponent proteins (PKSs) and long, biosynthetic metabolic pathways. 3) We will develop precursor biosynthetic pathways for precursors to polyketides and technologies to control pathway flux. 4) We will develop libraries of PKS modules and technology to mix-n-match these modules for production of very diverse chemicals, and demonstrate their utility for producing various acids and diacids. 5) We will develop computer aided design (CAD) software that will enable more rapid construction of yeasts for production of various chemicals, for predicting biosynthetic pathways for particular, desired chemicals, and for analyzing flux through metabolic pathways.