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菠菜中不同等电点的乙醇酸氧化酶的提取和特性
引用本文:杨净,徐杰,董宇亮,郭晋雅,曾秋莲,陆满芳.菠菜中不同等电点的乙醇酸氧化酶的提取和特性[J].华南师范大学学报(自然科学版),2005,0(4):91-98.
作者姓名:杨净  徐杰  董宇亮  郭晋雅  曾秋莲  陆满芳
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,广东广州,510631
基金项目: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)
摘    要:从不同批次的菠菜绿叶中可纯化出5种不同活性和等电点(pI)的乙醇酸氧化酶(CO).SDSPAGE表明它们均有40kD亚基,它们的比活会随蛋白浓度的增加而上升.第一种CO的pI〈8.3,比活为0.47U/mg,PAGE测定其全酶相对分子量(Mr)为280kD,280kD较稳定.第二种CO的pI〈8.3,但比活为3.44U/mg,PACE测定其Mr为560kD,但很快解聚为280kD.解释了前人报告CO的Mr大多为280kD.第三种CO的pI≈8.3,比活为8.4U/mg,放置后pI会下降至〈8.3,此时用PAGE测出其Mr为560kD.第四种CO的pI〉8.3,比活为27.8U/mg,放置pI也会下降为8.3.第五种CO的pI〉10.2,活性高达113U/mg,放置后其pI也会下降至〈8.3,由于CO的pI会下降,暗示低pI的CO可能并不存在于植物体内,而是实验中的娇作物.

关 键 词:乙醇酸氧化酶  纯化  pI  Mr  菠菜
文章编号:1000-5463(2005)04-0091-08
收稿时间:2005-06-06
修稿时间:2005年6月6日

PURIFICATION OF GLYCOLATE OXIDASE WITH DIFFERENT ISOELECTRIC POINTS FROM SPINACIA OLERACEA
YANG Jing,XU Jie,DONG Yu-liang,GUO Jin-ya,ZHENG Qiu-lian,LU Man-fang.PURIFICATION OF GLYCOLATE OXIDASE WITH DIFFERENT ISOELECTRIC POINTS FROM SPINACIA OLERACEA[J].Journal of South China Normal University(Natural Science Edition),2005,0(4):91-98.
Authors:YANG Jing  XU Jie  DONG Yu-liang  GUO Jin-ya  ZHENG Qiu-lian  LU Man-fang
Affiliation:College of Life Sciences, South China Normal University, Guangdong Key Lab of Biotechnology for Plant Development, Guangzhou , 510631 ,China
Abstract:Five kinds of glycolate oxidase(GO) with different pIs and specific activities were purified from green leaves of Spinacia oleracea.All of them contained(40 kD) subunit,their specific activities increased as protein concentration.One kind of GO displayed a specific activity of(0.47 U/mg),Mr of(280 kD),and a low pI(<8.3) assayed by PAGE.The second kind of GO had a pI<8.3,a specific activity of(3.44 U/mg) and a Mr of(560 kD).This GO was unstable and dissociate a product with Mr of(280 kD).Among them,GO with(280 kD) was stable that explained(280 kD) was always reported as the Mr of GO previously.The third kind of GO revealed a specific activity of(8.4 U/mg) and a pI about 8.3.This GO was labile and produced an artifact with pI<8.3 and Mr of(560 kD) after storage.The fourth kind of GO had a pI greater than 8.3 and a specific activity of(27.8 U/mg).Its pI also fell to about 8.3.The fifth kind of GO revealed a pI greater than 10.2 and a specific activity of(113 U/mg),its pI also decreased to the value lower than 8.3. In view of the decrease of pls after storage,it was most likely that not all of GO existed in plant indeed,some might be the artifact during the experiment,in despite of about ten kinds of GO with different pIs were reported previously.
Keywords:glycolate oxidase  purification  pI  Mr  spinach
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