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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
Source PublicationINTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume88Pages:88-92
SubtypeArticle
AbstractAllophycocyanin (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.
KeywordAllophycocyanin Escherichia Coli Thermostability Fluorescence
DOI10.1016/j.ijbiomac.2016.03.050
Indexed BySCI
Language英语
WOS IDWOS:000376800600011
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/126221
Collection实验海洋生物学重点实验室
Affiliation1.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
First Author AffilicationKey Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Recommended Citation
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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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