用液-固萃取体系从啤酒酵母粗提液中分离纯化葡糖-6-磷酸脱氢酶(英文)

Separation and Purification of Glucose 6 Phosphate Dehydrogenase from Rough Extract of Flesh Beer Yeast by Using Liquid Solid Extraction System *

在一定条件下，吐温８０的磷酸盐水溶液可分成聚合物固相和盐水相．酶、蛋白质等生物活性物质可在两相间进行分配．实验结果表明，吐温８０浓度和溶液ｐＨ的变化，以及提高盐浓度均会改变生物酶及蛋白质在两相中的分配．选择合适的条件，酶和蛋白质可获得分离，酶活力也得到纯化．研究了吐温８０浓度、盐浓度和溶液ｐＨ对啤酒酵母粗提液中葡糖－６－磷酸脱氢酶分配的影响．结果表明，选择吐温８０浓度１５％，总盐浓度３．０ｇ（Ｋ２ＨＰＯ４∶ＫＨ２ＰＯ４为５∶１），可得到好的分离结果．此时，一次纯化倍数１．７．酶活平均收得率为８９．０％．

Under given conditions, the aqueous solution of Tween 80 and phosphate can form polymeric solid phase and salt aqueous phase. Enzyme and protein, biological active material, can be distributed between the two phases. Experiment has shown that with the changes of Tween 80 concentration and pH of the solution, the increase of salt concentration, biological enzyme and protein will make some different distribution in the two phase. Under conditions chosen suitably , enzyme and protein can be separated and activity of enzyme also can be purified. In this paper, based on effect of Tween 80 concentration, salt concentration and pH on distribution of G 6 PDH from rough extract of flesh beer yeast have been investigated. The results indicated that appropriate Tween 80 concentration (15 %) and total salt weight (3.0 g ;K 2HPO 4 ∶ KH 2PO 4 = 5∶1) chosen can obtain better separation results. The purification fold is 1.7 a time. The average yield of enzyme activity is 89.0 %.