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

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

目的 利用三嗪类染料 Cibacron Blue F3 G-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 1 5 % (其中吐温 80中含染料配基吐温 80为 1 0 % ) ;成相磷酸钾盐浓度为 1 .49mol/L (p H 8.5 0 ) ;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 .91 mol/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 1 0 0 % ,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。

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.