用液相等电聚焦电泳纯化藻蓝蛋白亚基  

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

作　　者：黄峙[1] 杨芳[1] 郑文杰[1] 郭宝江[2] 

机构地区：[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

出　　处：《高等学校化学学报》2006年第6期1051-1054,共4页Chemical Journal of Chinese Universities

基　　金：广东省自然科学基金重点项目(批准号:05103295);教育部重点项目(批准号:01141)资助

摘　　要：以纯藻蓝蛋白(C-phycocyan in,C-PC)为材料,采用Rotofor系统进行液相等电聚焦(L iqu id-phase iso-eletric 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 eletro-phoresis)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 u ltra th in IEF)和SDS-PAGE电泳鉴定表明分别为高纯度的C-PCα,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.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 platertsis. After two steps of preand 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 a and fl subunits was 39.2%. The a and fl subunits of C-PC with a high purity as well as activity maintaining were obtained easily by this LP-IEF protocol, sugges- ting that it should be an ordinary but effective method for purification and preparation of protein subunits with different pI values.

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 

分 类 号：Q539[生物学—生物化学] O629[理学—有机化学]