The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project, if successful, will be to demonstrate the potential of the technology to dramatically reduce the cost and risk currently associated with production of bio-based products, enabling ultra-low cost product development. The field of metabolic engineering historically has been limited in predicting the behavior of complex biological systems in vivo from simplified models and basic in vitro biochemical principles. In many cases, it has proven much more difficult than expected to integrate a well characterized production pathway into a living host and balance the complex requirements of both biomass growth and production. The technology under development is a first of its kind, truly scalable, high-throughput metabolic engineering platform enabling the rapid development of microbial production strains. This significant reduction in development cost enables the possibility to produce numerous specialty products that would otherwise not have acceptable internal return on development capital.
This SBIR Phase II project will develop a high throughput metabolic engineering platform that enables the rapid development of microbial production strains. The platform, which bridges a gap between current in vivo and in vitro bio-production approaches, relies on the dynamic minimization of the active metabolic network in the context of a standardized two-stage bioprocess. Metabolic networks are highly interconnected wherein each metabolite and/or enzyme can interact with endless others. This combinatorial complexity results in a huge potential design space, which is intractable to the kinds of systematic experimentation required for the development of standardized design principles. The global challenges in addressing such a large biological design space have persisted, despite the dramatic advances in, and decreased costs of, reading and writing DNA, high-throughput DNA assembly, and microbial strain construction approaches. Dynamic metabolic network minimization not only results in a design space with greatly reduced complexity, but also provides strains that are robust to environmental conditions. Robustness leads to predictable scalability from high-throughput small-scale screens or "microfermentations" to fully instrumented bioreactors. This project will extend the validation of predictable strain performance from high-throughput microfermentation to pilot scale fermentation.