Abstract: Apostichopus japonicus spermary ( ASJS) was used as material to optimize enzymolysis technology according to the degree of hydrolysis, and antioxidant activities of hydrolysates were also investigated. The hydrolysis enzymes included PAP, PRO and FLA.The biuret method combined with TCA was used to determine degree of hydrolysis.Through single factor test and orthogonal test, the optimized conditions of each enzyme were obtained. And at the same time, the enzymolysis test used sole enzyme compared with two enzymes and three enzymes was carried out. Finally, antioxidant activities of different hydrolysates were determined in vitro.The results showed that the optimum conditions of enzymolysis technology with sole enzyme were as follows: PAP with the enzyme dosage 3.0%, temperature 70 ℃ and enzymolysis time 4 h, PRO with the enzyme dosage 3.0%, temperature 45 ℃ and enzymolysis time 4 h, FLA with the enzyme dosage 2.0%, temperature 45 ℃ and enzymolysis time 4 h.Under the optimum condition hydrolysis degrees of hydrolysis of PAP, PRO and FLA were 32.14%, 31.09% and 11.89%, respectively.The enzymolysis degrees of double enzyme and three enzymes were lower than sole PAP and PRO. The results of scavenging abilities of hydroxyl radical and superoxide anion radicals displayed that with concentrations of 5 mg/m L and10 mg/m L, the hydrolysate of ASJS hydrolyzed by PAP owned better ability than hydrolysates produced through PRO and FLA, double enzymes, three enzymes, and those from spawn and body wall hydrolyzed by PAP. In conclusion, PAP hydrolysate from ASJS exhibited good antioxidant activities, and it is promising to be utilized and exploited.