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Biosynthesis, spectral properties and thermostability of cyanobacterial allophycocyanin holo-alpha subunits
Chen, Huaxin1,2; Liu, Qiuzi3; Zhao, Jin1,2; Jiang, Peng1,2
2016-07-01
发表期刊INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷号88页码:88-92
文章类型Article
摘要Allophycocyanin (APC) is generally used as fluorescent labels. In this study, apcA genes from a mesophilic cyanobacterium Synechocystis sp. PCC 6803 and a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 were cloned into expression vectors to construct pathway for biosynthesis of allophycocyanin holo-alpha subunits (named as holo-ApcAS for Synechocystis sp. PCC 6803 and holo-ApcAT for T. elongatus BP-1) in Escherichia coli. The two holo-ApcAs were successfully reconstituted in E. coli and purified by metal affinity chromatography. Spectral analysis showed that the two proteins have similar spectroscopic properties, with absorbance maximum both at 614 nm and emission maximum at 639 nm for holo-ApcAS and 638 nm for holo-ApcAT. At high temperature, the recombinant holo-ApcAT was much more stable than the recombinant holo-ApcAS. Holo-ApcAS was most fluorescent at pH 8.5 and stable in pH range of 6.0-9.0 while holo-ApcAT was most fluorescent at pH 6.0 and stable in pH range of 5.0-7.0, with residual fluorescence intensity no less than 90% of the maximum fluorescence. These findings will pave the way for further protein engineering to achieve high stable APC from extremophiles. (C) 2016 Elsevier B.V. All rights reserved.
关键词Allophycocyanin Escherichia Coli Thermostability Fluorescence
DOI10.1016/j.ijbiomac.2016.03.050
收录类别SCI
语种英语
WOS记录号WOS:000376800600011
引用统计
被引频次:1[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/126221
专题实验海洋生物学重点实验室
作者单位1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
3.Qingdao Univ Sci & Technol, Qingdao 266071, Peoples R China
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Chen, Huaxin,Liu, Qiuzi,Zhao, Jin,et al. Biosynthesis, spectral properties and thermostability of cyanobacterial allophycocyanin holo-alpha subunits[J]. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,2016,88:88-92.
APA Chen, Huaxin,Liu, Qiuzi,Zhao, Jin,&Jiang, Peng.(2016).Biosynthesis, spectral properties and thermostability of cyanobacterial allophycocyanin holo-alpha subunits.INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,88,88-92.
MLA Chen, Huaxin,et al."Biosynthesis, spectral properties and thermostability of cyanobacterial allophycocyanin holo-alpha subunits".INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES 88(2016):88-92.
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