NaCl盐析提取酶解液中Ⅱ型胶原的工艺

Salt-out for extractingtype Ⅱ collagen from enzyme hydrolysate

采用响应面分析法对酶解液中Ⅱ型胶原的盐析条件进行优化,并通过SDS-PAGE电泳及圆二色谱对其纯度和结构进行鉴定。结果表明:酶解液中Ⅱ型胶原的最佳盐析条件为NaCl浓度3.5mol/L、22.3℃、31.08h,在此条件下Ⅱ型胶原回收率为90.88%;采用盐析法从酶解液中制备的Ⅱ型胶原,具有较高纯度并保持着完整的3股螺旋结构。

Response surface methodology was used to optimize technology of salt-out of type II collagen from enzymatic hydrolysate.SDS-PAGE and circular dichroism were used to determine the purity and the structure of type Ⅱ collagen.The results showed that the optimum parameters for NaCl salting out were as follows：extraction time 31.08 h,NaCl concentration 3.5 mol/L,temperature 22.3 ℃,and the recovery ratio 90.88%.Type II collagen with salting-out methodology is essentially free from contaminating proteins and keeps characteristic of triple helical conformation.