离子交换与吸附

以弱碱性大孔阴离子树脂DEAE-E/H为载体固定化氨基酰化酶。通过对影响固定化结果的几个因素,如树脂的离子类型、pH值、温度,以及自由酶液浓度等进行系统研究,得到了适宜的固定化条件:将DEAE-E/H转化为Ac-型;自由酶液浓度120U/ml,pH值6.5;固定化温度为常温。在此条件下制备的固定化氨基酰化酶比酶活可达1200U/g~1500U/g,酶活保留率超过60%。DEAE-E/H作为固定化载体,具有价格低廉,物理性能好,固定化方法简便等特点,具有很好的工业应用前景。

Diethylaminoethyl hydroethylate (DEAE-E/H), a kind of macroporous weakly basic anion exchanger, was used as carrier to immobilize aminoacylase. Several factors, such as the type of DEAE-E/H, pH, temperature, concentration of free enzyme solution, and so on, were investigated systematically. The results showed that the IME activity was influenced markedly by pH, while affected rarely by temperature in certain range. Finally, the appropriate immobilization conditions were obtained, which were: DEAE-E/H was transformed into the type of Ac-, pH was about 6.5, temperature was normal temperature, and the concentration of free enzyme solution was 120U/ml. Under the optimum conditions, the immobilized aminoacylase on DEAE-E/H was prepared and the specific activity was assayed. It illustrated that the specific activity was up to 1200U/g~1500U/g, and the activity yield was more than 60%. DEAE-E/H, as the carrier of immobilization, had such advantages as low cost, good physical property, and simple method of immobilization, therefore it had good industrial potentiality.