Metabolic Engineering of Escherichia coli for Production of Butyric Acid

Abstract

The butyrate production by engineered Escherichia coli is afflicted by both low titer and low selectivity (defined as the butyrate/acetate (B/A) ratio). To address this issue, a strategy for metabolic engineering of E. coli was implemented including (1) elimination of all major NADH-dependent reactions in the fermentation metabolism, (2) reconstruction of a heterologous pathway leading to butyryl-CoA, (3) recruitment of endogenous atoDA for conversion of butyryl-CoA to butyrate with acetate as a CoA acceptor, and (4) removal of the acetate-synthesis pathway. Grown on glucose (20 g/L) plus acetate (8 g/L), the engineered strain consumed almost all glucose and acetate and produced 10 g/L butyrate as a predominant product within 48 h. It leads to high butyrate selectivity with the B/A ratio reaching 143. The result shows that our proposed approach may open a new avenue in biotechnology for production of butyrate in E. coli.