仿刺参肠酶解条件的优化及其抗疲劳活性的研究

Study on the Optimization of Hydrolysis Conditions of Intestine of Sea Cucumber Apostichopus joponicus and Its Anti-fatigue Activity

以水解度为主要指标,筛选了碱性蛋白酶、中性蛋白酶、风味蛋白酶、复合蛋白酶、木瓜蛋白酶、胰蛋白酶、动物蛋白酶和水产蛋白酶对仿刺参肠的酶解效果,比较了p H、温度、时间、料液比和加酶量等单因素对水解度的影响,在单因素试验的基础上采用响应面试验设计对酶解条件进行了优化,并对最佳工艺下的水解产物高、中、低剂量对小鼠抗疲劳活性进行了研究。试验结果表明,水产蛋白酶对参肠蛋白的水解效果优于其它蛋白酶;响应面优化得到的最优水解条件为:温度50℃,酶的添加量2.3%,料液比1︰8.6(g/m L),p H 7.1,水解时间3.1 h,其水解度达到18.6%;参肠蛋白水解物的低中高剂量组均能显著延长小鼠的游泳时间,具有抗疲劳活性。

The degree of hydrolysis was taken as the main index to screen alcalase, neutrase, flavourzyme, protamex, papain, trypsin, animal protease and aquatic protease for hydrolysis of intestine of sea cucumber Apostichopus joponicus. Based on the effect of single factors, pH, enzymolysis temperature, hydrolyzation time, the ratio of material to solvent and dosage of enzyme on the degree of hydrolysis, the analysis of response surface method were carried out to optimize the hydrolysis process, and mice anti-fatigue activity of the hydrolysates at low, middle, high dosage obtained under optimal conditions were carried out. The results showed that the degree of aquatic protease hydrolysis was obviously higher than other enzymes tested. By the RSM transactional analysis, the optimum enzymolysis conditions were optimized as followings： enzymolysis temperature 50 ℃, the ratio of enzyme to substrate 2.3%, the ratio of material to solvent 1 ： 8.6 （g/mL）, pH 7.1 and time 3.1 h, respectively. The DH was up to 18.6% under the optimum conditions. All of the hydrolysates at low, middle, high dosage could significantly prolong the time of loaded-swimming, so it has significant anti-fatigue effect.