Site-specific immobilization of lysozyme upon affinity chromatography resin by forecasting lysine activity and controlling pH and epoxy group density
文献类型：期刊论文
| 作者 | Zhang, Yan; Zhang, Guifeng; Feng, Jing; Kong, Yingjun; Gao, Jianping; Li, Yang; Su, Zhiguo |
| 刊名 | JOURNAL OF CHROMATOGRAPHY A
 |
| 出版日期 | 2019-05-10 |
| 卷号 | 1592页码:192-196 |
| ISSN号 | 0021-9673 |
| 关键词 | Affinity chromatography resin Lysozyme Immobilization site HPLC-MS Molecular simulation |
| DOI | 10.1016/j.chroma.2019.02.066 |
| 英文摘要 | Here, a site-specific immobilization strategy for lysozyme was discussed. First, we calculated the surface micro-environment of lysozyme as a model protein and forecasted the nucleophilic attack activity order of six lysine residues within lysozyme, namely, K96> K97 and K33>K1 , K13, and K116. Second, lysozyme was immobilized on agarose resin (epoxy group density 22.34 mu mol/g) and incubated at pH 9.5 for 12 h. We then compared the peptide maps of free and immobilized lysozyme and established a quantitative detection method for immobilization sites. Third, we studied the effect of immobilization conditions such as epoxy group density and pH upon immobilization sites. When lysozyme was immobilized upon resin with a high epoxy group density (22.34 mu mol/g) at pH 9.5 for 12 h, multiple lysozyme immobilization sites were found, including K33, K96 and K97; when lysozyme was immobilized upon resin with a low epoxy group density (11.36 mu mol/g) at pH 9.5 for 12 h, only one lysozyme immobilization site was found, K96. The pH also had an effect upon immobilization sites. When immobilization was performed at pH >= 10.5 with low epoxy group density resin (11.36 mu mol/g) for 12 h, the immobilization sites included at least K33, K96 and K97; when immobilization was performed at pH 9.5 with other conditions remaining unchanged, the only lysozyme immobilization site found was at K96. These experimental results were highly consistent with the forecasted results, revealing some regularities of immobilization site control for lysozyme upon affinity chromatography resin. (C) 2019 Elsevier B.V. All rights reserved. |
| WOS关键词 | ORIENTED IMMOBILIZATION ; PROTEIN |
| 资助项目 | National Key Technology R&D Programs of China[2018YFA0108203] ; National Key Technology R&D Programs of China[2018YFC1106402] ; Guangzhou People's Livelihood Science and Technology Project of China[201803010086] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry |
| 语种 | 英语 |
| 出版者 | ELSEVIER SCIENCE BV |
| WOS记录号 | WOS:000465148200021 |
| 资助机构 | National Key Technology R&D Programs of China ; Guangzhou People's Livelihood Science and Technology Project of China |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/28204]  |
| 专题 | 中国科学院过程工程研究所 |
| 通讯作者 | Zhang, Guifeng |
| 作者单位 | Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, 1 Bei Er Tiao, Beijing 100190, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Yan,Zhang, Guifeng,Feng, Jing,et al. Site-specific immobilization of lysozyme upon affinity chromatography resin by forecasting lysine activity and controlling pH and epoxy group density[J]. JOURNAL OF CHROMATOGRAPHY A,2019,1592:192-196. |
| APA | Zhang, Yan.,Zhang, Guifeng.,Feng, Jing.,Kong, Yingjun.,Gao, Jianping.,...&Su, Zhiguo.(2019).Site-specific immobilization of lysozyme upon affinity chromatography resin by forecasting lysine activity and controlling pH and epoxy group density.JOURNAL OF CHROMATOGRAPHY A,1592,192-196. |
| MLA | Zhang, Yan,et al."Site-specific immobilization of lysozyme upon affinity chromatography resin by forecasting lysine activity and controlling pH and epoxy group density".JOURNAL OF CHROMATOGRAPHY A 1592(2019):192-196. |
入库方式： OAI收割
来源：过程工程研究所
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