灵芝TR6号漆酶的分离纯化及性质研究  

Purification and Characterization of Fungal Laccase from Ganoderma lucidum Strain TR6

作　　者：韩君莉[1] 郭丽琼[1] 郑晓冰[2] 周伟坚[2] 林俊芳[1] 

机构地区：[1]华南农业大学生物质能研究所,华南农业大学食品学院广州510642 [2]华南农业大学食品学院,广州510642

出　　处：《应用与环境生物学报》2008年第1期99-103,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(No30671457);广东省科技攻关项目(2003C31207,2004B20101001);广东省自然科学基金课题(No5006680)资助~~

摘　　要：采用硫酸铵盐析、DEAE Sepharose Fast Flow柱层析和Sephadex G100凝胶柱层析对灵芝TR6号漆酶发酵液进行分离纯化，得到电泳纯漆酶．SDS-PAGE电泳结果显示，该漆酶表观分子量为62×10^3．Native-PAGE鉴定该漆酶由一种同工酶组成，为单体酶．试验结果表明，该漆酶的最适反应温度为60℃，与ABTS的最适反应pH为4．5，Km为0．188mmol／L，在pH6．0～8．0之间较稳定．Fe^2＋、SDS和Tris对该漆酶活性具有抑制作用，EDTA和Tween80则对该漆酶活性具有促进作用．The laccase from Ganoderma lucidum strain TR6 was purified by ammonium sulfate salting-out, DEAE Sepharose Fast Flow column chromatography and Sephadex G100 gel column chromatography, and the electrophoretic pure laccase was obtained. The result of SDS - PAGE showed that the apparent molecular weight of the laccase was 62×10^3. The result of native -PAGE indicated that the laccasc was a monomeric protein with only one isoenzyme. Experimental results revealed that the optimum temperature and pH of the laccase reacted with ABTS were 60℃ and 4.5, respectively, and its Km value was 0. 188 mmol/L. The laccase was stable at pH ranging from 6.0 to 8.0. The laccase activity was strongly inhibited by Fe^2＋ , SDS and Tris, while enhanced by EDTA and Tween 80. Fig 8, Tab 2, Ref 10

关 键 词：灵芝 漆酶 纯化 酶活 

分 类 号：Q814[生物学—生物工程] S567.31[农业科学—中草药栽培]