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植物乙醇酸氧化酶分离纯化方法的改进
引用本文:郭晋雅,董宇亮,杨净,徐杰,尹汗萍,曾秋莲.植物乙醇酸氧化酶分离纯化方法的改进[J].植物学报,2006,23(6):703-707.
作者姓名:郭晋雅  董宇亮  杨净  徐杰  尹汗萍  曾秋莲
作者单位: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
摘    要:通过缩短DEAE-Cellulose柱长度, 加快流速并采用pH8.8的80 mmol.L-1 Tris-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(54.6~197.0 U.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40 kD带,表明3种植物乙醇酸氧化酶的亚基大小无区别。

关 键 词:乙醇酸氧化酶  纯化  菠菜  菜心  豆角
收稿时间:2006-03-13
修稿时间:2006-07-13

Improvement of the Purification Method of Plant Glycolate Oxidase
Jinya Guo,Yuliang Dong,Jing Yang,Jie Xu,Hanping Yin,Qiulian Zeng.Improvement of the Purification Method of Plant Glycolate Oxidase[J].Bulletin of Botany,2006,23(6):703-707.
Authors:Jinya Guo  Yuliang Dong  Jing Yang  Jie Xu  Hanping Yin  Qiulian Zeng
Affiliation:College of Life Sciences, South China Normal University, Guangdong Key Lab of Biotechnology for Plant Development, Guangzhou 510631, China
Abstract: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 pI (>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.
Keywords:,
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