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Site-specific immobilization of lysozyme upon affinity chromatography resin by forecasting lysine activity and controlling pH and epoxy group density
Zhang, Yan; Zhang, Guifeng; Feng, Jing; Kong, Yingjun; Gao, Jianping; Li, Yang; Su, Zhiguo
2019-05-10
Source PublicationJOURNAL OF CHROMATOGRAPHY A
ISSN0021-9673
Volume1592Pages:192-196
AbstractHere, 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.
KeywordAffinity chromatography resin Lysozyme Immobilization site HPLC-MS Molecular simulation
DOI10.1016/j.chroma.2019.02.066
Language英语
WOS KeywordORIENTED IMMOBILIZATION ; PROTEIN
Funding ProjectNational 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 Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
Funding OrganizationNational Key Technology R&D Programs of China ; Guangzhou People's Livelihood Science and Technology Project of China
WOS IDWOS:000465148200021
PublisherELSEVIER SCIENCE BV
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/28204
Collection中国科学院过程工程研究所
Corresponding AuthorZhang, Guifeng
AffiliationChinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, 1 Bei Er Tiao, Beijing 100190, Peoples R China
Recommended Citation
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.
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