Abstract: In order to extract total protein of Hippocampus and optimize conditions for enzymatic hydrolysis, we extracted the total protein of Hippocampus by the method of water extraction of dried Hippocampus. Papain, alkaline protease, neutral protease and trypsin were used respectively under the different pH, time, temperature and E/S conditions to conduct single factor test, and finally the enzymatic hydrolysis conditions were optimized by orthogonal test. The enzymatic hydrolysis condition and protein content under different conditions were determined by SDS-PAGE and BCA, respectively. The optimal enzymolysis conditions were determined according to the degree of proteolysis shown in the charts. The results showed that 1.056 g total protein was extracted from 10 g dried Hippocampus, and its protein concentration was 0.106 g/mL. The best enzyme was alkaline protease. The best enzymatic conditions were pH 9, E/S 5%, hydrolysis temperature 50℃and hydrolysis time 3 h, and the degree of hydrolysis of the Hippocampus protein was 96.9%. The optimal enzymatic hydrolysis conditions have been determined, and the study laid a certain foundation for the study of biological activity of hippocampal polypeptide.

Key words: Hippocampus, protein, hydrolysis conditions, optimization

摘要： 为了提取海马总蛋白并确定最佳的酶解条件，将海马粉碎后采用水提法提取海马总蛋白。分别选用木瓜蛋白酶、碱性蛋白酶、中性蛋白酶和胰蛋白酶，在不同酶解pH、时间、温度和E/S条件下进行单因素和正交试验法对海马总蛋白的酶解条件进行优化。利用聚丙烯酰胺凝胶电泳和BCA法来分别检测不同条件下的酶解情况及其蛋白含量，根据蛋白水解度来确定其最佳的酶解条件。实验结果表明：从10 g干海马中提取总蛋白为1.056 g，蛋白浓度为0.106 g/mL。海马总蛋白的最佳水解酶为碱性蛋白酶，其最佳酶解条件为：pH 9,E/S为5%,温度为50℃,时间为3 h，获得海马总蛋白的最大水解度为96.9%。研究获得了海马总蛋白及其最佳酶解条件，并为海马多肽生物活性研究奠定了基础。

关键词: 海马, 蛋白, 酶解条件, 优化