本研究以金钱龟肉糜为原料，通过可控的酶解技术制备抗肿瘤活性肽，研究中分别用胰蛋白酶、碱性蛋白酶、木瓜蛋白酶和复合蛋白酶protamex对金钱龟肉糜进行酶解并结合超滤初步分离得到了12种酶解产物，MTT实验表明其中胰蛋白酶酶解所得分子量小于3000 u的多肽E其抗肿瘤效果最强，当浓度为1 mg/mL时，对人肝癌细胞(HepG-2)和人乳腺癌细胞(MCF-7)的抑制率分别为92.95%和67.08%。将E利用SephadexG-15进一步纯化，经MTT实验筛选得到两组活性较强的多肽组份E1和E2，对HepG-2的抗增殖效果来看E1（IC50为77.29 ?g/mL）强于阳性对照5-氟尿嘧啶（IC50为136.27 ?g/mL），对于MCF-7的抗增殖效果来看E2(IC50为214.17 ?g/mL)同样要强于阳性对照5-氟尿嘧啶（IC50为245.82 ?g/mL）。本研究中，抗肿瘤活性肽首次从金钱龟中分离得出，对于以金钱龟为原料的抗肿瘤药物和保健品的开发具有重要指导意义。

Here, minced Cuora trifasciata meat was used as a raw material from which peptides with anti-tumor activities were prepared by controlled enzymolysis. Alcalase, papain, trypsin, and protamex were used to carry out the hydrolysis of minced Cuora trifasciata meat, and 12 different molecular weight hydrolyzates were obtained after ultrafiltration. A 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay showed that hydrolyzate E, produced from tryptic digestion with a molecular weight of <3000 u, exhibited the best anti-tumor effect. When the concentration was 1 mg/mL, the anti-proliferation rates of hydrolyzate E against human liver cancer cells (HepG-2) and human breast cancer cells (MCF-7) were 92.95% and 67.08%, rpesectively. Hydrolyzate E was further purified by Sephadex G-15 gel filtration, and two polypeptide fractions (E1 and E2) with strong activity were selected based on an MTT assay. E1 had a stronger anti-proliferative effect on HepG-2 cells (IC50 = 77.29 μg/mL) than the positive control 5-fluorouracil (IC50 = 136.27 μg/mL), while E2 had a stronger anti-proliferative effect on MCF-7 cells (IC50 = 214.17 μg/mL) than the positive control 5-fluorouracil (IC50 = 245.82 μg/mL). In this study, peptides with anti-tumor activity were isolated from Cuora trifasciata for the first time, which will have important implications in the development of anti-tumor drugs and health products using Cuora trifasciata as the raw material.

国家863科技计划项目子课题（2014AA022004）