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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

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【作者】 尹汉萍徐杰曾秋莲王再花叶庆生

【Author】 YIN Han- ping,XU Jie,ZENG Qiu - lian,WANG Zai-hua ,YE Qing- sheng(College of Life Science, South China Normal University, Guangdong Key Laboratory of Biotechnologyfor Plant Development, Guangzhou 510631, China; State Key Laboratory of Plant Physiologyand Biochemistry, China Agricultural University, Beijing 100094, China)

【机构】 华南师范大学生命科学学院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东广州 510631 中国农业大学植物生理学与生物化学国家重点开放实验室北京 100094广东省植物发育生物工程重点实验室广东广州 510631广东广州 510631

【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.

【Abstract】 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.

【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
  • 【文献出处】 华南师范大学学报(自然科学版) ,Journal of South China Normal University(Natural Science) , 编辑部邮箱 ,2004年03期
  • 【分类号】Q946
  • 【下载频次】282
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