Fumaric acid is one of the top 12-biomass building-block chemicals. In this study, we reported manipulation of E. coli central carbon metabolism with the aim to decrease the by-products and improve fumaric acid production. PEP-dependent glucose phosphotransferase system was replaced with a galactose translocation system to minimize the consumption of phosphoenolpyruvate. Engineering anaplerotic pathway (phosphoenolpyruvate carboxylase) was employed to redistribute carbon flux from glycolysis to Krebs cycle. Deletion of malate dehydrogenase and overexpression of acetyl-CoA synthase could decrease the byproducts malic acid and acetic acid. The combined strategies led to fumaric acid yield up to 1.53 g/g dry cell weight, a 50% increase compared with the parental strain. The result demonstrated that these genetic modifications were effective strategies for improving the production of fumaric acid and the engineered strain may serve a platform microbial cell factory for efficient production of fumaric acid or other dicarboxylic acids.
Keywords: Acetic acid; Escherichia coli; Fumaric acid; Malic acid; Phosphate-transferase system.
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