Development of robust, flexible and scalable multi-product platform microbes utilizing dynamic metabolic engineering

We have developed a robust and generalizable bioprocess platform, which decouples microbial growth and metabolic production. We are able to simultaneously eliminate combinations of key enzymes in central metabolism, minimizing the metabolic network and design space. Metabolic flux is effectively redirected from growth to production using synthetic metabolic “valves” which rely on a combination of controlled protein degradation and gene silencing, in a phosphate limited stationary phase. This methodology has been successfully demonstrated in the two important platform microbes, Escherichia coli and Saccharomyces cerevisiae. To date we have demonstrated the utility of this approach with numerous different production strains, resulting in the commercially relevant production of a wide variety of products ranging from biofuels, and commodity and specialty chemicals to natural products, and even lipidated peptides useful as biologics or in the production of nanostructures. Speed of strain optimization is increased due to the robustness of the minimal metabolic networks to key process conditions such as oxygen and glucose concentration. This robustness has enabled predictable scale up from screening scale (96 well plates) to controlled benchtop bioreactors.