利用双水相萃取技术分离纯化尿激酶

摘要

An aqueous two-phase extraction system using ethanol/(NH4)2 SO4 to separate urokinase was established.Factors of ethanol/(NH4)2SO4 ratio,pH,inorganic salt and crude enzyme concentration influencing the purification were investigated.The results showed that 65％ cold ethanol,10％ (NH4)2SO4,pH8.0,crude enzyme 30％.Under the optimal conditions,no inorganic salt was added.Purification urokinase was up to 9 times,and the recovery was 92％.%建立以乙醇/(NH4)2SO4组成的双水相萃取体系分离纯化尿激酶的新途径.通过研究双水相萃取系统的各种影响因素:乙醇/(NH4)2SO4的组成比例、pH、无机盐的加入、粗酶的浓度等,探索以乙醇/(NH4)2SO4组成的双水相萃取体系纯化尿激酶的最佳条件.研究结果表明:双水相萃取系统冰乙醇体积分数为65％,(NH4)2SO4质量分数为10％,pH 8.0,酶加入量为30％,且不加入任何其他无机盐的条件下,尿激酶的纯化倍数可达到9.0倍,回收率最高达92％.