Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag
Li, Wenjun1,3; Pu, Yang2; Gao, Na4; Tang, Zhihong1; Song, Lufei1; Qin, Song1
2017-03-01
发表期刊SAUDI JOURNAL OF BIOLOGICAL SCIENCES
ISSN1319-562X
卷号24期号:3页码:451-458
摘要Allophycocyanin plays a key role for the photon energy transfer from the phycobilisome to reaction center chlorophylls with high efficiency in cyanobacteria. Previously, the high soluble self-assembled bioengineering allophycocyanin trimer with N-terminus polyhistidine from Synechocystis sp. PCC 6803 had been successfully recombined and expressed in Escherichia coli strain. The standard protocol with immobilized metal-ion affinity chromatography with chelating transition metal ion (Ni2+) was used to purify the recombinant protein. Extensive optimization works were performed to obtain the desired protocol for high efficiency, low disassociation, simplicity and fitting for large-scale purification. In this study, a 3(3) full factorial response surface methodology was employed to optimize the varied factors such as pH of potassium phosphate (X-1), NaCl concentration (X-2), and imidazole concentration (X-3). A maximum trimerization ratio (Y-1) of approximate A(650 nm)/A(620 nm) at 1.024 was obtained at these optimum parameters. Further examinations, with absorbance spectra, fluorescence spectra and SDS-PAGE, confirmed the presence of bioengineering allophycocyanin trimer with highly trimeric form. All these results demonstrate that optimized protocol is efficient in purification of bioengineering allophycocyanin trimer with Histag. (C) 2017 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
关键词Efficient Purification Bioengineering Allophycocyanin Trimer Optimization Histag
DOI10.1016/j.sjbs.2017.01.011
项目资助者Science Foundation of the Chinese Academy of Sciences(XDA1102040300) ; Ocean Public Welfare Scientific Research Project, State Oceanic Administration of China(201205027) ; National Key Technology R&D Program of China(2013BAB01B01) ; Scientific Research Foundation of Ludong University(ly2014041)
收录类别SCI
关键词[WOS]ION AFFINITY-CHROMATOGRAPHY ; C-PHYCOCYANIN ; EXPERIMENTAL-DESIGN ; PHOTOSYSTEM-II ; OPTIMIZATION ; PEPTIDES ; ALPHA ; PHYCOBILISOME ; BIOSYNTHESIS ; EQUILIBRIUM
文章类型Article
语种英语
WOS研究方向Life Sciences & Biomedicine - Other Topics
WOS记录号WOS:000402474900001
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文献类型期刊论文
条目标识符http://ir.yic.ac.cn/handle/133337/22521
专题海岸带生物学与生物资源利用所重点实验室
作者单位1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China
2.Ludong Univ, Sch Agr, Yantai 264025, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Chinese Acad Sci, South China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China
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Li, Wenjun,Pu, Yang,Gao, Na,et al. Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag[J]. SAUDI JOURNAL OF BIOLOGICAL SCIENCES,2017,24(3):451-458.
APA Li, Wenjun,Pu, Yang,Gao, Na,Tang, Zhihong,Song, Lufei,&Qin, Song.(2017).Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag.SAUDI JOURNAL OF BIOLOGICAL SCIENCES,24(3),451-458.
MLA Li, Wenjun,et al."Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag".SAUDI JOURNAL OF BIOLOGICAL SCIENCES 24.3(2017):451-458.
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