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
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摘要: 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。Abstract: 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%,w