Engineering Clostridium acetobutylicum for Enhanced Solvent Production by Overexpression of Pyruvate Decarboxylase from Zymomonas mobilis

Clostridium acetobutylicum DSM 792 has been characterized as a native acetone-butanol-ethanol (ABE) fermenting bacterium. C. acetobutylicm is used as a model organism to improve solvent production by metabolic engineering strategies. In this study, the ABE pathway in C. acetobutylicum was modified by overexpressing the pyruvate decarboxylase gene from Zymomonas mobilis under the control of 2 promoters, namely Pferr and Ppta (promoters of genes encoding pyruvate ferredoxin oxidoreductase and phosphotransacetylase) and the recombinant strains were designated as DSM 792(pPDC1) and DSM 792(pPDC2). The specific activity of the alcohol dehydrogenases in the pdc-expressing strains was higher than that in the wild-type strain. In batch fermentation experiments, the total ABE production from the recombinant strains DSM 792 (pPDC1) and DSM 792 (pPDC2) was 1.57 and 7.9 g/L, respectively, which was higher when compared to the wild type ABE production (0.64 g/L) under uncontrolled pH. The alcohol to acetone ratio (BE/A) was found to be 2.28, 2.77, and 5.26 in wild type, DSM 792 (pPDC1), and DSM 792 (pPDC2), respectively. This suggests that the re-routing of pyruvate by overexpression of the pdc could play a role in the generation of more reducing equivalents towards ethanol and butanol production.