Abstract: To establish sea cucumber viscera hydrolysis conditions and provide a reference for the industrial production of sea cucumber viscera active substance. Degree of hydrolysis and clearing DPPH free radicals were selected as the indexes to compare the effect of hydrolysis and product antioxidant activity of neutral protease,alkaline protease,papain.Neutral protease was selected as the best protease.The hydrolysis conditions was optimized by response surface methodology.The results showed the optimal hydrolysis conditions: enzyme dosage 2000 U/g,material / liquid 1 ∶ 12 g/mL,hydrolysis temperature of 50 ° C,the hydrolysis conditions were pH7.0. Under this selected technological parameter,the effects of different extraction time on antioxidant activity were compared. Found that the hydrolyzate had good antioxidant activity at 10 h. Half inhibitory concentrations of DPPH radical,hydroxyl free radical were 1.26 and 5.33 mg/mL. Using a neutral enzymatic hydrolysis of sea cucumber offal process optimization was reasonable and feasible.Hydrolyzate has stronger antioxidant activity.