鱼腥藻PCC7120核膜连接蛋白ApcE体内重组研究

In vivo Reconstitution of Anabaena sp.PCC7120 Core-membrane Linker Protein ApcE with Phycocyanobilin

为了研究鱼腥藻PCC7120核-膜连接蛋白ApcE(1-240)脱辅基蛋白与藻蓝胆素的连接机制,通过体内重组方式得到色素蛋白PCB-ApcE(1-240)。吸收光谱、荧光光谱分析表明,核-膜连接蛋白ApcE(1-240)与藻蓝胆素进行了正确的体内重组。ApcE(1-240)脱辅基蛋白可与藻蓝胆素体内自催化共价连接,获得的色素蛋白溶于含有4 mol/L尿素的磷酸钾缓冲体系中,并具有最大吸收峰(λm ax=660 nm)和荧光发射峰(λm ax=668 nm)。

The deletion mutated core-membrane linker protein ApcE （1-240） trom Anabaena sp. PCC7120 were overexpressed in E. coli. The activities of expressed proteins were studied through in vivo reconstitution tests, where phycocyanobilin-biosynthesizing gene （ hol and pcyA） were transformed into E. coll. Through the absorption and the fluorescence spectra, the results show ApcE （1-240） could bind covalently phycocyanobilin （PCB） in an autocatalytic reaction. The presence of 4 mol/L urea is necessary to solubilize the resulting biliprotein, and it has absorption maxima （ λmax = 660 nm） and fluorescence maxima （ λmax = 668 nm）.