菠菜中高活性高等电点的乙醇酸氧化酶同工酶纯化方法的改进  

A MODIFIED METHOD FOR PURIFYING GLYCOLATE OXIDASE WITH HIGH ACTIVITY AND HIGH ISOELECTRIC POINT FROM GREEN LEAVES OF SPINACIA OLERACEA

作　　者：尹汉萍[1] 徐杰[1] 曾秋莲[1] 王再花[1] 叶庆生[1] 

机构地区：[1]华南师范大学生命科学学院

出　　处：《华南师范大学学报（自然科学版）》2004年第3期109-114,共6页Journal of South China Normal University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(39800009);广东省自然科学基金资助项目(021105);国农业大学植物生理学与生物化学国家重点开放实验室基金资助项目

摘　　要：通过缩短DEAE-celluloses柱长度,提高其洗脱液pH,可快速地从菠菜绿叶中纯化得到乙醇酸氧化酶同工酶,证实其具高活性113.0 U/mg·min和高等电点(pI)10.6.纯化后产率为0.14％,纯化倍数为66.5.经SDS-PAGE和毛细管电泳(CE-SDS)均只有40kD单带,说明已纯化至电泳纯.By shorting the length of DEAE-cellulose column and increasing the pH of elution solution, one kind of glycolate oxidase isozyme is purified rapidly from green leaves of spinach. Glycolate oxidase isozyme displays high specific activity about 113.0 U/mg · min protein and high electronic point (pI) of 10.6 assayed in acetate cellulose membrane electrophoresis under denaturing condition. A 66.5 - fold of purification and a yield of 0.14% are reached from the crude during this purification procedure. Only one band appeared when this enzyme preparation is adjusted to SDS - PAGE and to CE - SDS under denaturing condition, indicating this glycolate oxidase isozyme has been purified to apparent homogeneity.

关 键 词：菠菜 等电点 乙醇酸氧化酶同工酶 纯化 毛细管电泳 

分 类 号：Q55[生物学—生物化学]