The improvements of 1-hexadecylpyridinium chloride monohydrate on the production of glucose by the enzymatic hydrolysis of pure cellulose, the effects of enzyme activity and enzyme kinetics have been investigated. The results showed that, the optimal enzymatic hydrolysis conditions of pure cellulose were as follows: enzyme dosage was 10 FPU/g, mass fraction of substrate was 1.25%, surfactant mass concentration was 5g/L, reaction time was 12h. The average conversion rate of cellulose was 59.91%, and improved 20.79% compared with the value of 39.12% without adding surfactant. The addition of 1-hexadecylpyridinium chloride monohydrate increased the maximum rate of enzymatic hydrolysis and the Michaelis constant，and the maximum reaction rate increased by 58.22%, the Michaelis constant increased by 1.01 times. The relative enzyme activity increased firstly and then decreased with time, the highest value of relative enzyme activity increased was 53.18% at 24h.