仿刺参精酶解工艺条件优化及体外抗氧化

Optimization of enzymolysis technology of Apostichopus japonicus spermary and antioxidant activities in vitro of hydrolysates

本研究以仿刺参精(Apostichopus japonicus spermary,ASJS)为材料,以水解度为指标优化了酶解工艺条件,并考察了酶解产物的体外抗氧化效果。实验选用木瓜蛋白酶(PAP)、复合蛋白酶(PRO)和风味蛋白酶(FLA)作为实验用酶,以双缩脲法结合三氯乙酸法测定水解度,通过单因素与正交实验,优化了单酶酶解工艺条件,并与双酶及三酶复配水解进行了比较实验,最后对不同的水解产物进行了体外抗氧化活性测定。结果表明:单酶最佳酶解工艺条件为PAP:酶添加量3.0%、温度70℃及反应时间4 h;PRO:酶添加量3.0%、温度45℃及反应时间4 h;FLA为:酶添加量2.0%、温度45℃及反应时间4 h。在最佳酶解工艺条件下PAP水解度为32.14%,PRO为31.09%,FLA为11.89%。双酶及三酶的复配水解工艺检测到的水解度均低于单PAP及单PRO。不同水解产物清除羟自由基和超氧阴离子自由基能力结果为,在样品浓度为5 mg/m L和10 mg/m L时,ASJS的PAP酶解产物好于PRO、FLA、双酶、三酶复配,同时也好于仿刺参卵及体壁PAP酶解产物。综上可知,ASJS的PAP水解产物显示出较好的体外抗氧化活性,可以进一步开发利用。

Apostichopusjaponicus 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/mL and 10 mg/mL, 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.