Abstract: As the energy crisis intensifies worldwide, the development of cellulose has become an important method to solve the energy problem. This study is to improve the enzymatic hydrolysis efficiency of bamboo fiber material for technical parameters optimization. With sympodial bamboo after steam explosion pretreatment as raw material, a comprehensive investigation of 4 main factors, including temperature, pH, the amount of enzyme, beta-glucosidase/filter paper which affected the enzymatic hydrolysis of cellulose, was conducted to grasp bamboo enzymolysis optimizing conditions of various factors. An orthogonal optimization test was conducted with the four factors to achieve the goal of integration and optimization of enzymatic hydrolysis process of bamboo bioethanol. By analysis of the optimization test result, the optimum condition for enzymatic hydrolysis of bamboo was obtained as follows: reaction temperature was 50℃, initial pH was 5.2, substrate concentration was 10%, Tween-20 was 0.3%, the amount of enzyme was 35 IU/g, beta-glucosidase enzyme activity/ filter paper ratio was 1.0, hydrolysis time was 48 h. After fermentation condition was determined, fermentation amplified verification experiment was performed in 2.5 fermentor. 30.37 g/L of reducing sugars was generated, and the saccharification rate was 57.49%. The results suggest that the optimum condition established is useful and would benefit optimizing enzymatic catalytic hydrolysis and can provide scientific basis for bamboo fermentation process in the future.