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| 藻蓝蛋白裂合酶催化藻红胆素(PEB)与CpcA和PecA共价偶联 | |
| 引用本文： | 张举成,伍贤军,张娟,刘卫,周明.藻蓝蛋白裂合酶催化藻红胆素(PEB)与CpcA和PecA共价偶联[J].云南大学学报(自然科学版),2009,31(6):632-637. |
| 作者姓名： | 张举成 伍贤军 张娟 刘卫 周明 |
| 作者单位： | 1. 红河学院理学院, 云南蒙自 661100; 2. 华中科技大学环境科学与工程学院, 湖北武汉 430074; 3. 华中农业大学农业微生物国家重点实验室, 湖北武汉 430070; 4. 华中农业大学生物质与生物能源研究中心, 湖北武汉 430070 |
| 摘 要： | 将由鱼腥藻Anabaenasp.PCC7120中cpcE/F构建的pCOLADuet-cpcE/F质粒和pACYCDuet-pebA质粒、pCDFDuet-hol-pebB质粒、pETDuet-pecA质粒、pETDuet-cpcA质粒转化入大肠杆菌,在IPTG的诱导下,实现了色素蛋白PEB-CpcA和PEB-PecA的成功表达.吸收光谱和荧光光谱研究表明,CpcE/F能够催化PEB共价连接到CpcA和PecA的Cys-84上形成色素蛋白PEB-PecA和PEB-CpcA,色素蛋白的最大吸收峰为555nm,最强荧光峰为570nm.Zn2+电泳分析结果表明,体内重组获得了目标色素蛋白,证明由cpcE/F所编码的蛋白裂合酶在体内重组中能够催化PEB与CpcA和PecA的共价偶联反应,生成非天然色素蛋白PEB-CpcA和PEB-PecA. |
| 关 键 词： | 藻蓝蛋白α-CPC 藻红蓝蛋白α-PEC 裂合酶CpcE/F 体内重组 藻红胆素 |
| 收稿时间： | 2009-4-25 |
Phycoerythrobilin covalent coupled to CpcA and PecA with phycocyanin lyase from Anabaena sp. PCC 7120 |
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| ZHANG Ju-cheng,WU Xian-jun,ZHANG Juan,LIU Wei,ZHOU Ming.Phycoerythrobilin covalent coupled to CpcA and PecA with phycocyanin lyase from Anabaena sp. PCC 7120[J].Journal of Yunnan University(Natural Sciences),2009,31(6):632-637. | |
| Authors: | ZHANG Ju-cheng WU Xian-jun ZHANG Juan LIU Wei ZHOU Ming |
| Institution: | 1. College of Science, Honghe University, Mengzi 661100, China; 2. College of Envi.Science &; Eng., Huazhong University of Science and Technology, Wuhan 430074, China; 3. The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; 4. Biomass and Bioenergy Research Centre, Huazhong Agricultural University, 430070 Wuhan, China |
| Abstract: | The pCOLADuet-cpcE/Fplasmid,which expressed the phycocyaninlyase encoded by cpcE/Fin Anabaena sp.PCC7120,and other plasmids,including pACYCDuet-pebA,pCDFD uet-hol-pebB,pETD uet-pecAor pETD uet-cpcA,were transformed together in E.coli BL21(DE3).After being induced by IPTG,the chromoproteins PEB-CpcA and PEB-PecA were successfully expressed in E.coli.The Absorption and fluorescence spectra showed that CpcE/Fcould catalyse the covalent attachment of PEBat Cys-84 of α-subunit of phycocyanin and phycoerythrocyanin to form the chromoprotein PEB-PecAand PEB-CpcA,respectively.Themaximal Absorption of PEB-PecA and PEB-CpcA was 555 nm,and the maximal fluorescence was 570 nm.SDS-PAGE analysis displayed the Zn2+-induced fluorescence of the bound chromophore under irradiation by 280 nm,which showed that PEB-PecA and PEB-CpcA were got with the chromophorylation in E.coli.The results showed that the lyase CpcE/F could catalyse the covalent attachment of PEB to CpcAand PecA. |
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