| 植物乙醇酸氧化酶分离纯化方法的改进 |
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| 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].植物学通报,2006,23(6):703-707. |
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| 作者姓名： | 郭晋雅 董宇亮 杨净 徐杰 尹汗萍 曾秋莲 |
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| 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 |
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| 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。
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| 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 |
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| Jinya Guo,Yuliang Dong,Jing Yang,Jie Xu,Hanping Yin,Qiulian Zeng.Improvement of the Purification Method of Plant Glycolate Oxidase[J].Chinese Bulletin of Botany,2006,23(6):703-707. |
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| Authors: | Jinya Guo Yuliang Dong Jing Yang Jie Xu Hanping Yin Qiulian Zeng |
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| 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 purifi- cation 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. |
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| Keywords: | glycolate oxidase purification Spinacia oleracea Brassica parachinensis Phaseolus vulgaris |
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