温和条件下分离马胶鲨皮胶原蛋白的方法及其部分特性研究  

Studies of the characterization and isolation of collagen from scomberomorus ripho nins skins under some appropriate condition

作　　者：吴文惠[1] 姝亚[1] 包斌[1] 陆魏[1] 何培民[1] 张艳[1] 张兴群[2] 缪辉南[3] 曹张军[2] 

机构地区：[1]上海水产大学食品学院,上海200090 [2]东华大学生物工程系 [3]第二军医大学基础医学院生物化学和分子生物学教研室

出　　处：《中华航海医学与高气压医学杂志》2007年第3期133-136,共4页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：上海市重点学科建设项目;上海市教委重点研究项目资助(T1102;06412)

摘　　要：目的探讨分离马胶鲨皮胶原蛋白的方法以及马胶鲨皮胶原蛋白的部分生物化学特性。方法马胶鲨皮经清洗、浸泡、匀浆、离心得上清液。沉淀重复上述的提取过程,合并上清液,加硫酸铵沉淀胶原蛋白,回收的沉淀溶于磷酸盐缓冲液,经葡聚糖凝胶过滤后,蛋白溶出液被冷冻干燥。用聚丙烯酰胺凝胶电泳法、差示扫描量热仪法、圆二色谱仪法表征了通过上述磷酸盐缓冲液溶胀分离法获得的胶原蛋白的特性。结果用磷酸盐缓冲液溶胀分离法从马胶鲨皮分离得到胶原蛋白,胶原蛋白的回收率为鲜重的3.2%～3.4%;分离的胶原蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上呈现清晰的三条谱带,分子质量分别为205 000、134 000、118 000;马胶鲨皮胶原蛋白的最高热变性温度为47℃;分离的马胶鲨皮胶原蛋白在空间结构上以β-折叠为主,并包含有大量的无规卷曲。结论本研究所制备的高纯度胶原蛋白可用于生物医学材料的进一步研究。Objective To investigate the method of isolating collagen from scomberomorus ripho nins skins and to study its biochemical properties. Methods The properties of collagen from scomberomorus ripho nins skins were obtained by soaking,centrifuging, sedimentating,filtrating and frozen-drying. Then, the molecular weight, thermal denatured temperature and circular dichroism of purified collagen were determined by polyacrylamide gel electrophoresis, differential scanning thermo-equipment and circular dichroism chromatography. Results The results showed that fabricated collagen was composed of a high weight chain （205 000 ） and two low weight chains （134 000,118 000）. Its maximal thermal denature temperature was 47 ℃. The collagen from scomberomorus ripho nins skins was soaked in phosphate buffer and extracted in the solution, the recovery rate of which was 3.2% -3.4%. At the same time, the spatial structure of collagen were primarily β-fold and the random structure. Conclusions These findings suggest that the fabricated collagen can be served as a biomedical material for further research.

关 键 词：马胶鲨皮 胶原蛋白 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳 变性温度 二级结构 

分 类 号：R318.08[医药卫生—生物医学工程]