Molecular Design and Synthesis of a Novel Substrate for Assaying Lysozyme Activity.

IF 1.2 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of applied glycoscience Pub Date : 2018-08-20 eCollection Date: 2018-01-01 DOI:10.5458/jag.jag.JAG-2018_003

Megumi Matsui, Haruka Kono, Makoto Ogata

{"title":"Molecular Design and Synthesis of a Novel Substrate for Assaying Lysozyme Activity.","authors":"Megumi Matsui, Haruka Kono, Makoto Ogata","doi":"10.5458/jag.jag.JAG-2018_003","DOIUrl":null,"url":null,"abstract":"A novel substrate {Galβ1,4GlcNAcβ1,4GlcNAc-β-pNP [Gal(GlcNAc)2-β-pNP]} 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-pNP as the acceptor. Hydrolysis of the synthesized Gal(GlcNAc)2-β-pNP and related compounds using hen egg-white lysozyme (HEWL) demonstrated that the substrate was specifically cleaved to Gal(GlcNAc)2 and p-nitrophenol (pNP). 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-pNP selectively binds to a subsite of lysozyme to liberate the Gal(GlcNAc)2 and pNP products. The work therefore describes a new colorimetric method for quantifying lysozyme on the basis of the determination of pNP liberated from the substrate.","PeriodicalId":14999,"journal":{"name":"Journal of applied glycoscience","volume":"65 3","pages":"31-36"},"PeriodicalIF":1.2000,"publicationDate":"2018-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/bf/a7/JAG-65-031.PMC8056892.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of applied glycoscience","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.5458/jag.jag.JAG-2018_003","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2018/1/1 0:00:00","PubModel":"eCollection","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

A novel substrate {Galβ1,4GlcNAcβ1,4GlcNAc-β-pNP [Gal(GlcNAc)₂-β-pNP]} 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-pNP as the acceptor. Hydrolysis of the synthesized Gal(GlcNAc)₂-β-pNP and related compounds using hen egg-white lysozyme (HEWL) demonstrated that the substrate was specifically cleaved to Gal(GlcNAc)₂ and p-nitrophenol (pNP). 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-pNP selectively binds to a subsite of lysozyme to liberate the Gal(GlcNAc)₂ and pNP products. The work therefore describes a new colorimetric method for quantifying lysozyme on the basis of the determination of pNP liberated from the substrate.

Abstract Image

查看原文本刊更多论文

检测溶菌酶活性的新型底物的分子设计与合成

通过对接模拟设计了一种用于检测溶菌酶活性的新型底物 {Galβ1,4GlcNAcβ1,4GlcNAc-β-pNP [Gal(GlcNAc)2-β-pNP]}，并通过β-1,4-半乳糖基转移酶介导的转糖基化将 UDP-Gal 作为供体与 (GlcNAc)2-β-pNP 作为受体进行酶法合成。用母鸡卵白溶菌酶（HEWL）水解合成的Gal（GlcNAc）2-β-pNP和相关化合物的结果表明，底物被特异性地裂解为Gal（GlcNAc）2和对硝基苯酚（pNP）。为了阐明底物的结合模式，研究人员进一步将动力学研究和对接模拟相结合。结果表明，Gal(GlcNAc)2-β-pNP 可选择性地与溶菌酶的一个位点结合，从而释放出 Gal(GlcNAc)2 和 pNP 产物。因此，该研究描述了一种新的比色法，可根据从底物中释放出的 pNP 的测定结果对溶菌酶进行定量。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of applied glycoscience BIOCHEMISTRY & MOLECULAR BIOLOGY-

自引率

9.10%

发文量