双孢蘑菇多酚氧化酶的分离、纯化及特性分析

 通过分步盐析，DEAE-Cellulose-52阴离子交换层析和Sephadex G-100凝胶柱层析，对双孢蘑菇中的多酚氧化酶（PPO）进行了分离纯化，并对在分离纯化过程中蛋白得率及酶活性进行了测定。最终得到纯化倍数为103.2，比活为566.57 U · mg^-1，蛋白得率为0.47%的酶液。经SDS-PAGE电泳检验，所得到的PPO呈单一蛋白带，分子质量为25.5 kD。利用扫描探针显微镜对该酶的分子形态进行了观察，发现双孢蘑菇中的PPO分子呈椭球状，分子高度在8 ~ 12 nm之间。

Extraction,Purification and Property of Polyphenol Oxidase from Agaricus bisporus

Polyphenol oxidase(PPO)from Agaricus bisporus was extracted and purified by salting-out,DEAE-Cellulose-52 anion exchange chromatography and Sephadex G-100 gel column chromatography. The protein yield and PPO activity were analyzed during purification. The final purification fold,specific activity and protein yield were 103.2,566.57 U · mg-1 and 0.47%,respectively. The SDS-PAGE analysis showed that the PPO was a single protein band with a molecular weight of about 25.5 kD. The observation by scanning probe microscope showed that the PPO from Agaricus bisporus was ellipsoidal,with a diameter of about 8–12 nm.