植物乙醇酸氧化酶分离纯化方法的改进  

Improvement of the Purification Method of Plant Glycolate Oxidase

作　　者：郭晋雅[1] 董宇亮[1] 杨净[1] 徐杰[1] 尹汗萍[1] 曾秋莲[1] 

机构地区：[1]华南师范大学生命科学学院,广东省植物发育生物工程重点实验室,广州510631

出　　处：《植物学通报》2006年第6期703-707,共5页Chinese Bulletin of Botany

基　　金：广东省自然科学基金(No.021105)

摘　　要：通过缩短DEAE-Cellulose柱长度,加快流速并采用pH8.8的80mmol.L-1Tris-HCl为洗脱液,可在9小时内快速地从菠菜、菜心和豆角绿叶中纯化得到乙醇酸氧化酶。该酶具高活性(54.6～197.0U.mg-1)及高等电点(pI>10.0)。产率为4.1%～71.5%,纯化倍数为21.6～122.68。经SDS-PAGE检测均有40kD带,表明3种植物乙醇酸氧化酶的亚基大小无区别。By reducing the length of diethylaminoethanol-cellulose and increasing the pH in elution solution, we developed an improved method for purifying glycolate oxidase with high isoelectric value （pI 〉 10.0） from green leaves of Spinacia oleracea, Brassica parachinensis, and Phaseolus vulgaris. Purified glycolate oxidase showed high specific activity （54.6-197.0 U·mg^-1） and high pl （〉 10.0）. We achieved 21.6-122.68-fold purification and 4.1%-71.5% yield from crude solution during this purification procedure. SDS-PAGE of the glycolate oxidases revealed a 40-kD band, which indicates that the subunit molecular weight of glycolate oxidases in the 3 different plants was identical.

关 键 词：乙醇酸氧化酶 纯化 菠菜 菜心 豆角 

分 类 号：Q946[生物学—植物学]