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 αsubunit 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 (HistagPecF) were denatured and renatured together, and then the complex of PecE/HistagPecF was purified with Ni2+ chelating affinity chromatography. In this way, more active PecE/HistagPecF was obtained. In this enzymatic reconstitution, it is more safe that 2.5 mmol/L dithiothreitol (DTT) replaced 5 mmol/L βmercaptoethanol (ME), which is necessary cofactor in this enzymatic reaction.
出处
《华中师范大学学报（自然科学版）》
CAS
CSCD
北大核心
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).
