摘要
为得到中华草龟抗肿瘤活性肽的最佳提取工艺,选用胰蛋白酶、木瓜蛋白酶和复合蛋白酶对龟肉和龟血分别进行酶解单因素和正交试验,并依据其对白血病6T-CEM细胞株,肺癌A549细胞株,肝癌Hep G-2细胞株和乳腺癌MCF-7细胞株的抗癌活性,确定了木瓜蛋白酶的龟肉水解产物对乳腺癌MCF-7细胞株表现出较强的敏感性,再通过正交试验的极差分析与方差分析得出龟肉最佳酶解工艺条件为pH值7.5、酶解温度60℃、加酶量(E/S)0.5%、料液比1∶3,此时抑癌率达到80%左右。对酶解时间对酶解产物的影响也进行了研究,得出酶解时间在8h时,龟肉酶解产物的抗肿瘤活性较强。此外,质谱结果也进一步表明,酶解8h所得小分子肽产物的丰度最大,这为后续抗肿瘤生物活性肽的分离纯化奠定了良好基础。
In order to obtain the optimal extraction technology of antitumor bioactive peptides of Chinemys reevesii, and trypsin, papain and compound protease were chose to conduct enzymatic hydrolysis single factor and orthogonal experiment with tortoise meat and tortoise blood, respectively. Acccording to anticancer activity of hydrolysis products on leukemia 6T - CEM cell, lung cancer A549 cell, liver cancer HepG - 2 cell and breast cancer MCF - 7 cell, tortoise meat hydrolysis products of papain showed stronger sensitivity on breast cancer MCF -7 cell. The optimum enzymolysis technology condition of tortoise meat was obtained through range analysis and variance analysis of orthogonal experiment. The results showed that pH 7.5, hydrolysis temperature 60℃ , enzyme dosage （E/S） 0.5% , solid - liquid ratio 1 ： 3 and the cancer - inhibition rate could reach 80%. The effect of enzymolysis time on enzymolysis products was also studied. When enzymolysis time was 8h, tortoise meat enzymolysis products showed stronger antitumor activity. In addition, the results of mass spectrum also indicated that when enzymolysis time was 8h, the abundance of small molecular peptides products was the maximum, the good foundation for further separation and purification of antitumor bioactive peptides was established.
出处
《肉类工业》
2017年第7期28-33,共6页
Meat Industry
基金
安徽省水产产业技术体系项目(皖农科【2016】84号)
关键词
中华草龟
抗癌
活性肽
酶解
Chinemys reevesii
anticarcinogenic bioactive peptides
enzymatic hydrolysis
