Increasing cellulosic ethanol production by enhancing phenolic tolerance of Zymomonas mobilis in adaptive evolution.

Cellulosic ethanol fermentability of ethanologenic strain Zymomonas mobilis is severely inhibited by phenolic aldehydes generated from lignocellulose pretreatment. Here, a 198 days' laboratory adaptive evolution of Z. mobilis 8b in corn stover hydrolysate was conducted to increase its phenolic aldehydes tolerance and ethanol fermentability. The obtained Z. mobilis Z198 demonstrated a significantly improved conversion of the most toxic phenolic aldehyde (vanillin) by 6.3-fold and cellulosic ethanol production by 21.6%. The transcriptional analysis using qRT-PCR revealed that the gene ZMO3_RS07160 encoding SDR family oxidoreductase in Z. mobilis Z198 was significantly up-regulated by 11.7-fold. The overexpression of ZMO3_RS07160 in the parental Z. mobilis increased the ethanol fermentability to that of the adaptively evolved strain Z. mobilis Z198. This study provided a practical method to obtain a robust cellulosic ethanol fermenting strain, and a candidate gene for synthetic biology of biorefinery strains with strong phenolic aldehydes tolerance.