AbstractWheat straw is a suitable raw material for ethanol production since it has high cellulose content. The objective of this work was to evaluate the effect of cellobiase and surfactant on the enzymatic hydrolysis of lignocellulosic materials. For this purpose, wheat straw was first pretreated by organosolv digestion. The chemical s of raw and pretreated wheat straw were analyzed. Much of the hemicellulose and lignin were removed, and the relative cellulose content of pretreated wheat straw was 26.57% higher when compared to untreated wheat straw. Cellobiase was added into hydrolysate to improve the hydrolysis efficiency. Through experiments and analysis, the optimum cellobiase dosage was found to be 1/10 of the cellulase loading. Surfactant was also added into hydrolysate. Nonionic surfactant (Tween 80) exhibited better effect on improving enzymatic hydrolysis. When 0.06 g/g dry solids (DS) Tween 80 was also added into hydrolysate, the yield of glucose in hydrolyzate could reach 486 g/kg DS.