Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag
Li, WJ; Pu, Y; Gao, N; Tang, ZH; Song, LF; Qin, S; cpra_yic@sina.com
2017
发表期刊SAUDI JOURNAL OF BIOLOGICAL SCIENCES
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.
部门归属[Li, Wenjun; Tang, Zhihong; Song, Lufei; Qin, Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China; [Pu, Yang] Ludong Univ, Sch Agr, Yantai 264025, Peoples R China; [Li, Wenjun] Univ Chinese Acad Sci, Beijing 100049, Peoples R China; [Gao, Na] Chinese Acad Sci, South China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China
关键词Efficient Purification Bioengineering Allophycocyanin Trimer Optimization Histag
资助项目LMB
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/16253
专题中科院海洋生物资源可持续利用重点实验室
通讯作者cpra_yic@sina.com
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GB/T 7714
Li, WJ,Pu, Y,Gao, N,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, WJ.,Pu, Y.,Gao, N.,Tang, ZH.,Song, LF.,...&cpra_yic@sina.com.(2017).Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag.SAUDI JOURNAL OF BIOLOGICAL SCIENCES,24(3),451-458.
MLA Li, WJ,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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