This Small Business Innovation Research (STTR) Phase I project aims to develop a novel, high flux terpenoid precursor pathway by circumventing limitations of the bacterial methyl erythritol-phosphate (MEP) pathway for the renewable production of monoterpenoids. Monoterpenoids are natural chemical precursors for several consumer products, and many are produced via highly polluting chemical processes. In the proposed project the plan is to sidestep some of the MEP pathway limitations by designing de novo metabolic pathways. The designed/predicted enzymes will be characterized individually and assembled into a pathway. Further, multivariate-modular metabolic engineering (MMME) approaches will be used to assemble the upstream and downstream pathways to optimize the metabolic flux for the overproduction at commercially viable levels.
The broader impact/commercial potential of this project, if successful, will be to develop a microbial monoterpenoid production platform from renewable sugars that will retain and develop sustainable manufacturing of monoterpenoid-derived products in the US. By this strategy, terpenoids can be made at much higher productivities than the native bacterial MEP pathway. While the immediate focus is on the $1B+ monoterpene/derivative market, this approach will benefit US manufacturing of all terpenoids, in total a $5B+ market. Overall, this project will provide a new sustainable production route for these natural chemicals.
Through this NSF STTR Phase I funded research project, we have successfully completed the identification of a de novo pathway for the production of the terpenoid precursors IPP and DMAPP in E. coli. We have optimized expression and functional activities for several heterologous enzymes by evaluating different expression platforms and cofactor dependencies. Several candidate enzymes have been identified which catalyze unique reactions for the biosynthesis of novel intermediates to IPP and DMAPP. Through this work, we will dramatically expand our capabilities for microbial production of terpenes in E. coli. Continued application of our multivariate modular metabolic engineering techniques and protein engineering tools will serve to create a platform for the biosynthesis of a wide variety of terpene chemicals used in consumer products, pharmaceuticals, and biofuel applications. In striving to accomplish these goals, we have also developed a suite of new internal expertise and technologies which we have begun to and will continue to disseminate throughout the scientific community in the form of peer reviewed publications and well-trained research scientists, technicians, and managers, thereby strengthening the industrial biotechnology sector in the United States. As this project progresses through funding from grants and other funding sources, it will continue to create new jobs for a skilled workforce. This NSF STTR Phase I research project has not only enabled the translation of our academic research into an industrially-validated technology, but has also dramatically increased the probability of its commercialization.
