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目的优选长牡蛎肉的中性蛋白酶、木瓜蛋白酶、菠萝蛋白酶三酶复合水解工艺,通过动物实验研究其水解物的抗疲劳效果。方法以水解度为考察指标,采用正交实验优选长牡蛎肉三酶复合水解的最佳工艺;水解物用昆明种小鼠试验,观察负重游泳时间、耐缺氧能力,测定抗疲劳相关生化指标,根据实验结果判断抗疲劳效果。结果影响长牡蛎肉水解效果的主要因素是酶用量(P<0.05);中性蛋白酶、木瓜蛋白酶、菠萝蛋白酶最佳条件分别为:pH7.0、6.5、7.0;温度55、55、50℃;料水比1∶4、1∶4、1∶8;酶用量4 000、4 000、12 000 U.g-1;时间3、3、5 h。不连续聚丙烯酰胺凝胶垂直板电泳结果表明,水解物主要成分相对分子质量(Mr)300~1 000。与对照组相比,长牡蛎肉水解物高剂量组小鼠耐缺氧能力明显升高(P<0.01)、运动后血乳酸及血清尿素氮含量明显降低(P<0.01)、肝脏组织超氧化物歧化酶(SOD)活力明显升高(P<0.01),中、高剂量组小鼠肝糖原含量明显升高(P<0.01)、负重游泳时间明显延长(P<0.01)、红细胞(Hb)SOD活力明显升高(P<0.01)。结论长牡蛎肉三酶复合水解物经动物实验证明具有较好的抗疲劳效果。
OBJECTIVE To study the hydrolysis process of neutral protease, papain and bromelain in long-oyster meat and to study the anti-fatigue effect of hydrolyzate by animal experiments. Methods The degree of hydrolysis was taken as the index of investigation. Orthogonal experiment was carried out to optimize the compound enzyme hydrolysis process of long-oyster meat. Hydrolyzate was used in Kunming mice to observe the weight-bearing swimming time, hypoxia tolerance and anti-fatigue related biochemical indexes , According to the experimental results to determine the anti-fatigue effect. Results The main factors influencing the hydrolysis of long-oyster meat were enzyme dosage (P <0.05). The optimum conditions of neutral protease, papain and bromelain were pH7.0, 6.5 and 7.0, temperature 55, 55 and 50 ℃, Water ratio of 1: 4,1: 4,1: 8; enzyme dosage 4 000,4 000,12 000 Ug-1; time 3, 3, 5 h. Vertical polyacrylamide gel electrophoresis results show that the main components of the hydrolyzate relative molecular mass (Mr) 300 ~ 1000. Compared with the control group, the hypoxia tolerance of the high-dose oyster meat hydrolyzate high-dose group was significantly increased (P <0.01), and the levels of serum lactate and serum urea nitrogen were significantly decreased (P <0.01) (P <0.01). The contents of hepatic glycogen in the medium and high dose groups were significantly increased (P <0.01), the weight-bearing swimming time was significantly longer (P <0.01), and the levels of red blood cells ) SOD activity was significantly increased (P <0.01). Conclusion The results showed that the three-enzyme hydrolyzate of long-oyster meat had good anti-fatigue effect by animal experiments.