鱿鱼软骨Ⅱ型胶原蛋白提取方法及结构分析

Extracting Technology and Structure Characterization of Type Ⅱ Collagen from Squid Cartilage

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)Na OH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,Na Cl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 k Da,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 itsstructure. Methods Freeze - dry squid cartilage and pulverize them to 200 mesh powder, pretreat the powderwith 4 % （ w /w ） NaOH solution and 0. 5 M EDTA solution to remove polysaccharides and deliming, and then pepsinhydrolys, NaCl dialyzed with purified water and freeze - dried are used to obtain the type II collagen. The collagenwas 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 threea l 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.