Abstract: 【Objective】 To optimize the condition parameters of enzymatic hydrolysis and detoxification of cottonseed meal by gossypol degrading enzyme. 【Method】 Through the orthogonal design of 4 factors and 4 levels, the optimum conditions of hydrolysis of cottonseed meal were selected with the content of free gossypol as the index. 【Result】The optimum conditions for the detoxification of cottonseed meal by gossypol-degrading enzyme were as follows: Temperature 35℃, time 1.5 h, inoculation amount 3%, the substrate moisture 35%. 【Conclusion】The selected enzymatic detoxification parameters have significant effects. Under the conditions of optimal hydrolysis parameters, it was verified by experiments that the content of free gossypol in cottonseed meal decreased from 341.02 mg/kg to 37.01 mg/kg, and the degradation rate of free gossypol was 89.15%, which could be applied to the enzyme-catalyzed detoxification of free gossypol in cottonseed meal.