藻红蓝蛋白α-亚基体外重组体系的优化  

Improvement of reconstitution in vitro of phycoerythrocyanin α-subunit

作　　者：周明[1] 武栋[2] 赵开弘[2] 

机构地区：[1]华中科技大学环境科学与工程学院,武汉430074 [2]华中科技大学生命科学与技术学院,武汉430074

出　　处：《华中师范大学学报（自然科学版）》2002年第4期494-497,共4页Journal of Central China Normal University：Natural Sciences

基　　金：国家自然科学基金资助项目(30270326和90201001).

摘　　要：对我们建立的藻红蓝蛋白α-亚基体外重组体系进行了优化.实验表明:让表达的不具有组氨酸亲和标记的藻红蓝蛋白α-亚基连接/异构酶E亚基(简称PecE)与具有组氨酸亲和标记的藻红蓝蛋白α-亚基连接/异构酶F亚基(简称Histag-PecF)共同变性再复性,然后通过Ni2+金属鳌合亲和层析提纯PecE/Histag-PecF复合物,该PecE/Histag-PecF具有更好的催化活性.Reconstitution in vitro of phycoerythrocyanin αsubunit was improved further in this work. E subunit of phycoerythrocyanin lyase/isomerase with no histidine affinity tag (PecE) and F subunit of phycoerythrocyanin lyase/isomerase with histidine affinity tag (HistagPecF) were denatured and renatured together, and then the complex of PecE/HistagPecF was purified with Ni2+ chelating affinity chromatography. In this way, more active PecE/HistagPecF was obtained. In this enzymatic reconstitution, it is more safe that 2.5 mmol/L dithiothreitol (DTT) replaced 5 mmol/L βmercaptoethanol (ME), which is necessary cofactor in this enzymatic reaction.

关 键 词：α-亚基 体外重组体系 藻红蓝蛋白 连接/异构酶 基因重组 藻类植物 色素蛋白 

分 类 号：Q784[生物学—分子生物学] Q949.2