用三嗪染料配基吐温80—磷酸盐液固萃取体系分离纯化乳酸脱氢酶

目的：利用三嗪类染料Cibacron Blue F3G-A修饰的吐温80，与磷酸钾盐构建染料配基吐温80－盐液－固亲和萃取体系，从猪心肌匀浆液中分离纯化乳酸脱氢酶。方法：成相染料配基吐温80浓度为15％（其中吐温80中含染料配基吐温80为10％）；成相磷酸钾盐浓度为1．49mol/L(pH8.50);匀浆液加入量为体系的1％，一次成相正萃取。将吐温80相保温溶解，改变磷酸钾盐浓度为0．91mol/L,二次成相反萃取。结果：一次成相正萃取吐温80相平均酶活收得率97％，纯化8．0倍。二次成相反萃取，盐水相酶活收得率近100％，实现了酶与吐温80相的分离。结论：三嗪染料配基吐温80－磷酸盐液－固萃取体系,分离纯化乳酸脱氢酶，具有良好选择性和分离纯化效果。对其进一步研究与开发，具有较大实际应用价值。

Partitioning of Lactate Dehydrogenase from Porcine Cardia-muscle Based on Triazine Modified-Tween80-K_2HPO_4/KH_2PO_4 Liquid-Solid Extraction

ABSTRAC The partitioning of lactate dehydrogenase(LDH) from porcine cardia muscle using triazine modified Tween80 K 2HPO 4/KH 2PO 4 liquid solid extraction was studied. Under the appropriate separation conditions, the Conc. of Tween 80 was 15%,with triazine dye Tween 80 in which 10%, the Conc. of phosphate, 1.49 mol/L, pH 8.50, the homogenization added to the system,0.1 ml. The enzyme recovery in Tween80 phase was 97%, and the purification fold was 8.0. With the Conc. of phosphate adjusted to 0.91 mol/L, the dissolved Tween 80 re formed the liquid solid two phase system, and the enzyme recovery in salt aqueous phase was 99%, which simplified the following processing of LDH.