For efficient bioethanol production from lignocellulosic biomass by Saccharomyces cerevisiae, it is necessary to improve cellular tolerance to toxic compounds released during the pretreatment of biomass. The gene encoding p-nitrophenylphosphatase, PHO13, was disrupted in a recombinant xylose-fermenting S. cerevisiae strain, which improved ethanol production from xylose in the presence of three major inhibitors, acetic and formic acids, and furfural. In medium supplemented with 30 mM acetic acid, the ethanol yield obtained by the Delta PHO13 mutant was 0.45 g-ethanol/g-xylose. Notably, the specific ethanol productivity of the mutant in the presence of 90 mM furfural was fourfold higher than that of the control strain. The PHO13-disrupted strain produced ethanol from rice straw hydrolysate obtained by liquid hot-water pretreatment with a greater than fourfold higher xylose consumption rate than the control. Together, our findings demonstrate that PHO13 deletion is a simple, but effective, approach for improving cellulosic bioethanol production by S. cerevisiae. (C) 2012 Elsevier Ltd. All rights reserved.