酶解工艺对Ⅱ型胶原蛋白分子结构及提取率的影响

The Research of Purification and Characteration of TypeⅡ Collagen

以鸡胸软骨为原料制备Ⅱ型胶原蛋白,在酸性条件下添加胃蛋白酶酶解鸡胸软骨Ⅱ型胶原三股螺旋链的端肽,采用园二色谱研究酶解时间、温度及胃蛋白酶浓度对Ⅱ型胶原二级结构及提取率的影响,从而确定酶解工艺,并通过原子力显微镜及红外光谱对Ⅱ型胶原的分子结构进行表征。结果表明,酶解时间、温度对Ⅱ型胶原二级结构有显著影响,在胃蛋白酶作用温度为20℃,反应时间为32h,浓度为1%时,Ⅱ型胶原保持着比较完整的三股螺旋结构和较高的提取率。

This paper reported the preparation of type Ⅱ collagen from chick sternal cartilage. Pepsin was applied to digest the telopeptide of type Ⅱ collagen, and the effect of time, temperature and pepsin concentration on the secondary structure of type Ⅱ collagen were researched using circular diehroism. Atomic Force Microscope and FT-IR were applied to identify the structure of type Ⅱ collagen. The results suggest that time and temperature cause significant impact on the secondary structure of type Ⅱ collagen. Type Ⅱ collagen of higher extraction rate and better stability were obtained with the extraction time of 32h and 1% pepsin concentration at 37℃.