鸡关节软骨Ⅱ型胶原的制备

Preparation of Pure TypeⅡCollagen from Chicken Articular Cartilage

本文以鸡关节软骨为原料制备Ⅱ型胶原。在酸性条件下添加不同浓度的胃蛋白酶酶解鸡关节软骨Ⅱ型胶原三股螺旋链的端肽,使Ⅱ型胶原由不溶性转变为可溶性,研究底物粘度随酶解时间的变化,确定最佳的酶浓度及酶解时间。采用DEAE-Sephalose CL 6B离子交换柱和去离子水重复洗涤两种方法对酶解样品进行纯化制得Ⅱ型胶原,研究表明,胃蛋白酶最适添加浓度1%(W/W),最佳酶解时间24h。两种纯化方法的Ⅱ型胶原提取率分别为67%和81%。SDS-PAGE电泳、高效液相色谱、氨基酸成分分析及紫外吸收光谱测定结果证实了由鸡关节软骨所制备的Ⅱ型胶原具有较高纯度。

This paper reported the preparation of the type Ⅱ collagen from chicken articular cartilage. The type Ⅱ collagen was digested by various pepsin concentrations in acid solution. By investigating viscosity changes of type Ⅱ collagen, the optimal pepsin concentration and digestion time have been obtained in this process. To get the purer type Ⅱ collagen, DEAE-sephalose CL 6B column chromatographer was applied and repeatedly deionized in water elution. The results suggested that the optimal digestion conditions are 1% pepsin for 24 h. The extracton rate of type Ⅱ collagen by two methods are 67% and 81% respectively. Analysis results of SDS-PAGE electrophoresis, RP-HPLC, amino acid components and violet chromatography showed that the prepared type Ⅱ collagen by this method is a high purity product.