鱿鱼软骨II型胶原蛋白提取方法及结构分析

摘要

目的：从鱿鱼软骨中分离纯化非变性II型胶原蛋白，并对其进行结构表征。方法将鱿鱼软骨冻干后粉碎成200目软骨粉，用4%（w/w）NaOH溶液去除多糖、0.5 M EDTA溶液脱灰对软骨进行前处理，采用胃蛋白酶水解提取，NaCl盐析用纯水透析后冷冻干燥得到II型胶原蛋白样品。通过氨基酸组成分析、SDS-PAGE电泳、紫外吸收光谱、电镜扫描等对制备的样品鉴定。结果制备的II型胶原蛋白具有3条α1链，分子量112 kDa，在224.5 nm波长处有最大紫外吸收，具有三螺旋结构特征。%Objective Extract and purify native type II collagen from squid cartilage,and characterize its structure. Methods Freeze-dry squid cartilage and pulverize them to 200 mesh powder,pretreat the powder with 4%(w/w)NaOH solution and 0. 5 M EDTA solution to remove polysaccharides and deliming,and then pep-sin hydrolys,NaCl dialyzed with purified water and freeze-dried are used to obtain the type II collagen. The colla-gen was characterized by amino acid composition analysis,SDS-PAGE electrophoresis,UV absorption spectra, and scanning electron microscopy. Results The type II collagen prepared from squid cartilage possesses threeα1 chain,and the molecular weight of it is 112 kDa. In addition,it shows maximum UV absorption at 224. 5 nm, and performs triple helix characteristic.