高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

用液相等电聚焦电泳纯化藻蓝蛋白亚基

黄峙1,2, 杨芳1, 郑文杰1, 郭宝江2   

  1. 1. 暨南大学生命科学技术学院, 广州 510632; 2. 华南师范大学生命科学学院, 广州 510631

  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-06-10 发布日期:2006-06-10
  • 通讯作者: 郭宝江

Purification and Preparation of Active Subunits of C-phycocyanin by a Protocol of Liquidphase Isoeletric Focusing

HUANG Zhi1,2, YANG Fang1, ZHENG Wen-Jie1, GUO Bao-Jiang2*   

  1. 1. Department of Biotech, Life Science College, Jinan University, Guangzhou 510632, China;
    2. Biotechnology Research Institute, South China Normal University, Guangzhou 510631, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-06-10 Published:2006-06-10
  • Contact: GUO Bao-Jiang

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被引次数

28

摘要： 以纯藻蓝蛋白(C-phycocyanin, C-PC)为材料, 采用Rotofor系统进行液相等电聚焦(Liquid-phase isoeletric focusing, LP-IEF)电泳纯化C-PC的α, β亚基, 探讨蛋白质亚基纯化的制备电泳(Preparative eletrophoresis)技术. 结果显示, 样品经2次等电聚焦电泳后, C-PC 的α, β亚基分别浓集在pH=4.9和pH=4.1附近, 平板超薄等电聚焦(Slab ultra thin IEF)和SDS-PAGE电泳鉴定表明分别为高纯度的C-PC α, β亚基. 提示LP-IEF是分离纯化等电点差异蛋白质活性亚基的简便有效的方法.

关键词: 蛋白质亚基, 液相等电聚焦电泳, 等电点, 藻蓝蛋白, 制备电泳

Abstract: A process of liquidphase isoeletric focusing(LP-IEF) was developed and conducted in a 8.0 mol/L urea medium in Rotofor system for isolation, purification and preparation the α and β subunits of C-PC(αβ)3 purified from Spirulina platensis. After two steps of pre- and re- fractionating, collected fractions with α or β subunit were identified and characterized by SDSPAGE and slab ultra thin IEF. Its absorption and fluorescence spectra were determined. The results show that the α and β subunits were concentrated at about pH=4.9 and pH=4.1 respectively and the total recovery of α and β subunits was 392%. The α and β subunits of C-PC with a high purity as well as activity maintaining were obtained easily by this LP-IEF protocol, suggesting that it should be an ordinary but effective method for purification and preparation of protein subunits with different pI values.

Key words: Protein subunit, Liquidphase preparative IEF(LP-IEF), Isoelectric point(pI), C-phycocyanin(C-PC), Preparative electrophoresis

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