双孢蘑菇多酚氧化酶的分离、纯化及特性分析  

Extraction,Purification and Property of Polyphenol Oxidase from Agaricus bisporus

作　　者：朱继英[1] 王健[1] 王相友[1] 

机构地区：[1]山东理工大学农业工程与食品科学学院,山东淄博255049

出　　处：《园艺学报》2012年第4期763-768,共6页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30871757);国家农业科技成果转化资金项目(2010GB2C600253)

摘　　要：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566.57U·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25.5kD。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8~12nm之间。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.

关 键 词：双孢蘑菇 多酚氧化酶 纯化 分子形态 

分 类 号：S646.11[农业科学—蔬菜学]