Efficient purification protocol for bioengineering allophycocyanin trimer with N-terminus Histag
文献类型：期刊论文
| 作者 | Li, WJ; Pu, Y; Gao, N; Tang, ZH; Song, LF; Qin, S |
| 刊名 | SAUDI JOURNAL OF BIOLOGICAL SCIENCES
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| 出版日期 | 2017 |
| 卷号 | 24期号:3页码:451-458 |
| 关键词 | Efficient purification Bioengineering allophycocyanin trimer Optimization Histag |
| 通讯作者 | cpra_yic@sina.com |
| 中文摘要 | 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. |
| 源URL | [http://ir.scsio.ac.cn/handle/344004/16253]  |
| 专题 | 南海海洋研究所_中科院海洋生物资源可持续利用重点实验室 |
| 推荐引用方式 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,&Qin, S.(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. |
入库方式： OAI收割
来源：南海海洋研究所
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