用染料修饰吐温80液—固萃取体系从猪心肌匀浆液中分离纯化心肌黄酶

用三嗪类染料 Ｃｉｂａｃｒｏｎ Ｂｌｕｅ Ｆ３Ｇ－Ａ修饰的吐温８０，与吐温８０、硫酸被构成液－固萃取体系，从猪心肌匀浆液中分离纯化心肌黄酶。研究了吐温８０染料修饰物在吐温８０相中所占的比例、分相盐浓度、溶液的酸度、匀浆液的加入量等对匀浆液中酶及杂蛋白在两相中分配的影响。在室温条件下，酶选择性地进入吐温８０固相，杂蛋白主要留在盐水相。匀浆液中心肌黄酶的酶活力平均收得率为８１．４％，一步纯化倍数为６．６。降低盐浓度，提高盐水相酸度，能使酶从吐温８０固相反萃到盐水相。

SEPARATION AND PURIFICATION OF DIAPHORESES FROM HOMOGENATE OF PORK HEART MUSCLE USING LIQUID-SOLID EXTRACTION SYSTEM OF TWEEN 80 MODIFIED BY TRIAZINE DYE

The separation and purification of diaphorase from the homogenate of pork heart muscle was carried out using the liquid-solid extraction system consisted with Tween 80 modified by triazine dye Cibacron Blue F3G-A,Tween 80 and sulfate. The content of Tween 80 modified by dye in the Tween 80 phase,the salt concentration for forming two phases, the solution acidity and the amounts of ho- mogenate were tested for their effect on the distribution behavior between two phases of the enzyme and other proteins. Under the condition of indoor temperature,the enzyme can be extracted selectively into Tween 80 solid phase and a large of other proteins remains in the salt aqueous phase. The average activity recovery of diaphorase from the homogenate is 81. 4 %,and purification fold for one time is 6. 6. A large of diaphorase can be reverse extracted into the salt aqueous phase from the Tween 80 solid phase by decreasing the salt concentration and increasing the acidity of the salt aqueous phase.