用三嗪染料修饰吐温80液-固萃取乳酸脱氢酶的研究

Partitioning of Lactate Dehydrogenase from Porcine Cardia-Muscle Based on Triazine Dye-Tween 80-(NH_4)_2SO_4 Liquid-Solid Affinity Extraction System

利用三嗪类染料 Cibacron Blue F3G- A修饰吐温 80 ,与硫酸铵构建染料聚合物 -盐的液 -固亲和萃取体系 ,从猪心肌匀浆液中分离纯化乳酸脱氢酶。结果表明 ,吐温 80浓度 12 % ,含染料吐温 80为 5 % ;硫酸铵浓度1.81mol/ L ;p H 7.4 ;匀浆液加入量为体系的 1%时 ,吐温 80相平均酶活收率 93% ,纯化倍数 9.2。研究了乳酸脱氢酶与吐温 80相分离的条件 ,将吐温 80相保温溶解 ,改变硫酸铵浓度为 0 .9m ol/ L ,二次成相反萃 ,盐水相酶活收率90 % 。

The partitioning of lactate dehydrogenase (LDH) from porcine cardia muscle using triazine dye Tween 80 (NH 4) 2SO 4 liquid solid extraction was studied. The optimal conditions for extraction of LDH were obtained. The concentration of phase forming Tween 80 was 12%, containing 5% of triazine dye Tween 80. The concentration of phase forming salt, (NH 4) 2SO 4, was 1.81 mol/L, and pH 7.4. When the percentage of porcine cardia muscle homogenization added to the system was 1%, the target enzyme was extracted to the Tween 80 phase. The enzyme recovery and the purification were 93% and 9.2 fold, respectively. The separation of LDH from the polymer phase was also studied. When the concentration of (NH 4) 2SO 4 was adjusted to 0.9 mol/L, the dissolved Tween 80 could reform liquid solid two phase system and the enzyme recovery in salt aqueous phase was 90%. Thus the following process might be simplified.