Abstract: Biological-alkaline/oxidative pretreatment（BAO pretreatment）with white rot fungi is a promising technology for cellulosic ethanol production because of the advantages of mild environmental conditions and low energy consumption. In order to obtain the optimal conditions for enzymatic hydrolysis of corn straw treated by BAO pretreatment, the effects of cellulase loading, reaction time and substrate concentration on reducing sugar yields were evaluated by dynamics study, and the reaction temperature, rotational speed and pH value of enzymatic hydrolysis were also optimized by response surface design. The results indicated that 30 FPU/g of cellulase loading, 20 g/L of substrate concentration, 49℃ of reaction temperature, 200 r/min of rotational speed and pH 4.8 for 48 hours could be the most effective and economical way of enzymatic hydrolysis. Under such conditions, the reducing sugar yield reached (0.479±0.012)g/g .