牛肝脏透明质酸酶提取工艺优化

Optimization of extraction process of hyaluronidase from bovine liver

以牛肝脏为原料,确定分离纯化牛肝脏透明质酸酶的最优条件。实验采用Plackett-Burman设计、最陡爬坡实验、响应面中心旋转组合实验确定的最佳提取条件为:乙酸-乙酸钠浸提液pH4.69,NaCl浓度0.22mol/L,提取时间为10.82h,该条件下牛肝脏透明质酸酶比活力为33.27U/g。采用硫酸铵盐析和SephacrylS-100初步纯化,得到牛肝脏透明质酸酶比活力为452.81U/g。

As the raw material,hyaluronidase was isolated from bovine liver,the optimistic conditions was investigated. Plackett -Burman experiment,steepest ascent experiment and Central composite design were performed. The results showed that the optimistic isolation parameter were pH4.69 of acetate buffer,0.22mol/L NaCl and extraction time 10.82h. Under these conditions,the specific activity of hyaluronidase from bovine liver reached 33.27U/g. By ammonia sulfate salting-out and Sephacryl S-100 preliminary purification,the specific activity reached 452.81U/g.