This Small Business Innovation Research Phase I project will develop a custom-designed microbial biocatalyst for the renewable production of high value terpenoid biochemicals. Terpenoid biochemicals derived from essential oils are used in numerous consumer products and as food additives. Many of them accumulate in nature in various stereo-isomeric forms, each of which possesses unique properties and applications. These molecules are believed to function principally in ecological roles, serving as herbivore-feeding deterrents, antifungal defenses, and pollinator attractants. The research objective is to develop a fermentation process for biosynthetic production allowing increased adoption of such natural alternatives to synthetic chemicals. Multivariate-Modular Metabolic engineering (MMME) approaches will be used to transfer the natural biosynthetic pathway from the plant to a bacterial host and to optimize the metabolic flux for the overproduction at a commercially viable level. A high-productivity strain is anticipated, suitable for continued commercialization efforts. Overall, this project, if successful, will provide a new sustainable production route for these natural chemicals.

The broader impact/commercial potential of this project is the development of a microbial process for the economic and sustainable production of high value terpenoid biochemicals. These terpenoid biochemicals have applications in a variety of industries ranging from agro-chemicals, petro-chemicals and flavor and fragrance (F&F) chemicals; specifically they are commonly used as flavor agents, bio- herbicides, sprout inhibitors and as bio-pest repellents. In all, the total potential addressable markets exceed $3 Billion. Microbial production will benefit society by improving the renewability of the production process and by relocating production from overseas to the US. In summary, the development of microbes capable of producing the target will enable sustainable production of the target as well as create jobs in the US. This research will develop generalizable microbial strain engineering techniques for the high-volume production of natural products through a sustainable manufacturing process.

Project Report

Through this NSF SBIR Phase I/IB funded research project we have successfully completed proof-of-principle validation of our platform technology, specifically towards biosynthesis of a high value biochemical from terpene chemical family. We have identified optimal heterologous enzymes for biosynthesis in E. coli and searching through several orthologous enzymes and their functional expression produced the desired product. Our efforts identified unique complexities arising through the effects of pathway balancing on product accumulation which clearly shows that our MMME pathway optimization is key for improving the terpene titer in E. coli. Furthermore, we validated novel capabilities for biosynthesis of hydroxylated terpene product in E. coli which will dramatically expand our capability to address new and interesting chemical markets. Continued application of our multivariate modular metabolic engineering techniques and protein engineering tools will serve to create a platform for biosynthesis of a wide variety of high value terpene chemicals used in consumer product and pharmaceutical applications. In striving to accomplish these goals we've developed 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 commercial partners, it will continue to create new jobs for a skilled workforce. This NSF SBIR 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.

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Manus Biosynthesis, Inc.
United States
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