文章基本信息
- 标题：Molecular Design and Synthesis of a Novel Substrate for Assaying Lysozyme Activity
- 作者：Megumi Matsui ; Haruka Kono ; Makoto Ogata 等
- 期刊名称：Journal of Applied Glycoscience
- 印刷版ISSN：1344-7882
- 电子版ISSN：1880-7291
- 出版年度：2018
- 卷号：65
- 期号：3
- 页码：31-36
- DOI：10.5458/jag.jag.JAG-2018_003
- 语种：English
- 出版社：The Japanese Society of Applied Glycoscience
- 摘要：
A novel substrate {Galβ1,4GlcNAcβ1,4GlcNAc-β- p NP [Gal(GlcNAc)2-β- p NP]} for assaying lysozyme activity has been designed using docking simulations and enzymatic synthesis via β-1,4-galactosyltransferase-mediated transglycosylation from UDP-Gal as the donor to (GlcNAc)2-β- p NP as the acceptor. Hydrolysis of the synthesized Gal(GlcNAc)2-β- p NP and related compounds using hen egg-white lysozyme (HEWL) demonstrated that the substrate was specifically cleaved to Gal(GlcNAc)2 and p -nitrophenol ( p NP). A combination of kinetic studies and docking simulation was further conducted to elucidate the mode of substrate binding. The results demonstrate that Gal(GlcNAc)2-β- p NP selectively binds to a subsite of lysozyme to liberate the Gal(GlcNAc)2 and p NP products. The work therefore describes a new colorimetric method for quantifying lysozyme on the basis of the determination of p NP liberated from the substrate.
- 关键词：lysozyme;enzyme assay;kinetics;chitin oligosaccharide derivatives;enzymatic synthesis
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