We have developed a robust and scalable Two-Stage fermentation system, which decouples microbial growth and product formation. Dynamic metabolic control was used to automatically and effectively redirect metabolic flux from growth to production by synthetic metabolic “valves” using a combination of controlled protein degradation and CRISPR interference, in a phosphate limited stationary phase. The Two-Stage fermentation system is robust across a broad range of process conditions, under different oxygen and glucose conditions, and is capable of faithful scale up from µL scale in 96-well plate to L scale fermentors. Two-Stage fermentation was compared to growth-associated fermentation with the former consistently yielding more coherent results and is more robust across a wider spectrum of process conditions. We have demonstrated efficient production of fuels, commodity chemicals and candidate pharmaceutical products at high titers using the Two-Stage fermentation system.<br/>

Development and demonstration of a robust and scalable two-stage fermentation system for enhanced bioproduction

253rd National Meeting (2017)

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