SBIR Phase II: Novel Proteolysis-based Tools for Metabolic Engineering

This Small Business Innovation Research (SBIR) project aims to engineer microbes for the cost-effective production of specialty chemicals. Currently, engineered microbial strains bear mutations that increase the production of chemicals of interest by inhibiting the cell's ability to produce off pathway chemicals. These "loss-of-function" mutations are critical as they effectively channel the cell's metabolic flux toward the product of interest. This both boosts the production efficiency and eases downstream purification by eliminating the accumulation of undesirable but chemically-similar contaminants. Unfortunately, these mutations may also decrease the fitness of the cells and, as a result, the growth media must be supplemented with costly nutrients. Technical research herein will assess the feasibility of applying novel regulated proteolysis technology to simultaneously direct maximal metabolic flux toward the target chemical of interest while avoiding the need to supplement the growth media. If successful, this technology would provide a great cost savings and enable fermentative production to be applied more broadly in the production of specialty chemicals. The broader impact/commercial potential of this project is to provide a stable and cost-effective fermentative production route to a specialty chemical. Fermentative production of chemicals offers many advantages over traditional petrochemical or extraction-based production processes. Petrochemical production maintains the nation?s reliance on an unsustainable feedstock (oil) and also leads to national security issues as the US is largely dependent on foreign oil sources. Chemical production via extraction from plant materials also has ecological challenges. The process often uses toxic solvents, and may rely on unsustainable farming practices for many plants that are not traditional food crops. Engineered microbes fermented on sugar feedstock produced using high-efficiency agricultural practices offer a stable alternative for producing specialty chemicals, both in terms of supply and price.