Engineering Escherichia coli for improved ethanol production from gluconate

• Deletion of pflA and ldh led to significant improvement of ethanol yield on gluconate.
• Deletion of pflA and ldh led to small improvement of ethanol yield on glucose.
• Deletion of pdh alone led to improved ethanol yield on gluconate.
• Deletion of pdh alone led to no improvement of ethanol yield on glucose.

We report on engineering Escherichia coli to produce ethanol at high yield from gluconic acid (gluconate). Knocking out genes encoding for the competing pathways (l-lactate dehydrogenase and pyruvate formate lyase A) in E. coli KO11 eliminated lactate production, lowered the carbon flow toward acetate production, and improved the ethanol yield from 87.5% to 97.5% of the theoretical maximum, while the growth rate of the mutant strain was about 70% of the wild type. The corresponding genetic modifications led to a small improvement of ethanol yield from 101.5% to 106.0% on glucose. Deletion of the pyruvate dehydrogenase gene (pdh) alone improved the ethanol yield from 87.5% to 90.4% when gluconate was a substrate. The growth rate of the mutant strain was identical to that of the wild type. The corresponding genetic modification led to no improvements on ethanol yield on glucose.