大肠杆菌AE109青霉素G酰化酶的分离纯化及性质研究

The Purification and Properties of Penicillin G Acylase From E.coli AE109

由发酵培养液所得大肠杆菌AE109菌体,先经高渗休克处理,继经D-苯甘氨酸-Sepharose 4B和DEAE-纤维素柱层析分离纯化得到青霉素G酰化酶,酶制品在非变性条件下的聚丙烯酰胺凝胶电泳上呈一条区带,而且可以结晶。在SDS变性条件下解离为α和β两个亚基。 酶性质的研究结果表明,由大肠杆菌工程菌AE109菌株所得青霉素G酰化酶与其亲本大肠杆菌AS1.76菌株所得青霉素G酰化酶性质相同。

Penicillin G acylase has been isolated and purified from E.coli AE109 by treating with high osmotic shock followed by hydrophobic interaction chromatography and DEAE-cellulose chromatography .Electrophoresis analysis shown that the enzyme preparation has a high purity and can be crystallized. The enzyme was dissociaertd into subunit α and β on denaturing SDS-PAGE.Studies on the enzyme properties shown that the penicillin G acylase from E, coli AE109 strain is identical with the enzyme from one of the parent strain E. coli As 1.76.