固定化酶催化合成头孢克洛与产物的分离纯化

Synthesis and Purification of Cefaclor with Immobilized Enzyme

应用来源于粪产碱杆菌的青霉素G酰化酶催化合成头孢克洛,并对生物转化反应过程进行了研究。在反应条件:pH 7.0、25℃、磷酸钠缓冲体系、加酶量3 U/mL和底物浓度60 mmol/L时,转化率达到85%,表明粪产碱杆菌青霉素G酰化酶具有较高的催化活力。研究了新型分离纯化方法,经过树脂分离的产物纯度超过95%,主要杂质含量小于0.5%,可以达到药典要求,为生物法合成头孢抗生素走向产业化奠定了基础。

Penicillin G acylase from Alcaligenes faecalis （AfPGA） was used for the biosynthesis of cephalosporins. Optimization of the reaction was accomplished and the optimal results were achieved in phosphate buffer of pH 7.0 at 25 ℃, with substrate concentration of 60 mmol/L and enzyme of 3 U/mL. Conversion of substrate reached 85%. The results proved that AfPGA was eximious on enzyme activity. Isolation method was studied and resin was applied for separation. The purity of product reached 95 % and the main by-product was less than 0.5 %. The results were according with the pharmacopoeia and made the biotransformation process possible for industrial application.