An extra copy of the β-glucosidase gene improved the cellobiose fermentation capability of an engineered strain.

In a previously engineered recombinant, the cellobiose fermentation rate was significantly lower than the glucose fermentation rate. Thus, we implemented a genome-wide perturbation library to find gene targets for improving the cellobiose fermentation capability of the yeast strain. Unexpectedly, we discovered a transformant that contained an additional β-glucosidase gene (-), possibly through homologous recombination between the plasmids. The additional β-glucosidase led to the fastest cellobiose fermentation activity among all the transformants evaluated, and the strain demonstrated significantly higher β-glucosidase activity than the control strain, especially during the initial exponential growth phase. The present work revealed the benefit of the extra - copy for efficient cellobiose fermentation in the engineered strain.