鸡胸软骨中Ⅱ型胶原的制备工艺优化

Preparation Technology of Collagen Type Ⅱ from Chicken Sternal Cartilage

对鸡胸软骨Ⅱ型胶原的制备工艺进行优化。采用盐酸胍和Na Cl 2种抽提方法去除杂蛋白,然后用胃蛋白酶酶解并考察酶解温度、酶解时间和酶添加量对胶原提取率的影响。结果表明:4 mol/L的盐酸胍能有效去除杂蛋白;当酶解温度18℃、胃蛋白酶添加量1.59%、酶解43 h时提取率最高,且此条件下的胶原提取率为56.54%,且保持着完整的二级结构。经过酶解后制备的Ⅱ型胶原电泳图谱出现α、β2条带,无其他杂带,Ⅱ型胶原纯度较高。

This paper reported the preparation of the type Ⅱ collagen from chicken articular cartilage. Guanidine hydrochloride and Na Cl were used to remove the contaminating proteins, and the collagen was then digested with pepsin. The study investigated effects of reaction temperature, reaction time and concentration of enzyme on extraction efficiency. The results showed that guanidine hydrochloride at 4 mol/L removed heteropolysaccharide effectively. The optimal conditions for collagen type Ⅱ extraction were as follows: temperature, 18 ℃; gastric protein concentration, 1.59%; and digestion time, 43 h. Under these conditions, the extraction efficiency of collagen was 56.54%.