Abstract

Butanol toxicity, oxygen sensitivity, and high substrate cost limit wider applications of the traditional Clostridium acetone-butanol-ethanol (ABE) fermentation. In this study, a wild-type Clostridium beijerinckii strain was partnered with an engineered Escherichia coli strain to improve the ABE fermentation through a co-culture. Butanol and butyrate produced by C. beijerinckii were converted into butyl butyrate – a product that can be in situ removed to reduce product toxicity – by expressing a CoA transferase and an alcohol acyltransferase in E. coli. When integrated with a pretreatment technology and using commercial cellulase, the co-culture produced 1280 mg/L butyl butyrate from rice straw in bioreactors, without the need of maintaining a strict anaerobic condition. As a process harnessing both the genetic tractability of E. coli and the superior acid- and alcohol-producing ability of Clostridium, this co-culture fermentation will be useful in producing valuable esters from low-cost waste streams.

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