{"title":"perdeutero - N-sulfoheparosan C5-epimerization的1H NMR分析:一种直接测量固定化C5-epimerase活性的方法","authors":"Meng Qiao , Ruoyu Jiao , Xing Zhang , Zhengzhang Wu , Baoxing Shen , Bingzhi Li","doi":"10.1080/07328303.2021.1903026","DOIUrl":null,"url":null,"abstract":"<div><p>D-Glucuronyl C5-epimerase (C5-epi), which converts D-glucuronic acid to L-iduronic acid, is a crucial enzyme in the preparation of bioengineered heparin, which provides an alternative way to produce safer heparins. Herein, we provided an efficient way to measure the activity of cyanogen bromide-immobilized C5-epi based on proton nuclear magnetic resonance (NMR) analysis of perdeutero <em>N</em>-sulfoheparosan C5-epimerization. In this method, reactions catalyzed by C5-epi could be monitored by real-time 1 D NMR according to the peak intensity change of H-5 from IdoA and GlcA residues in the <sup>1</sup>H NMR spectrum, which is nondestructive of sample and also serves the kinetic property evaluation of immobilized C5-epi.</p></div>","PeriodicalId":1,"journal":{"name":"Accounts of Chemical Research","volume":null,"pages":null},"PeriodicalIF":16.4000,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/07328303.2021.1903026","citationCount":"1","resultStr":"{\"title\":\"1H NMR analysis of perdeutero N-sulfoheparosan C5-epimerization: a direct way to measure the activity of immobilized C5-epimerase\",\"authors\":\"Meng Qiao , Ruoyu Jiao , Xing Zhang , Zhengzhang Wu , Baoxing Shen , Bingzhi Li\",\"doi\":\"10.1080/07328303.2021.1903026\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>D-Glucuronyl C5-epimerase (C5-epi), which converts D-glucuronic acid to L-iduronic acid, is a crucial enzyme in the preparation of bioengineered heparin, which provides an alternative way to produce safer heparins. Herein, we provided an efficient way to measure the activity of cyanogen bromide-immobilized C5-epi based on proton nuclear magnetic resonance (NMR) analysis of perdeutero <em>N</em>-sulfoheparosan C5-epimerization. In this method, reactions catalyzed by C5-epi could be monitored by real-time 1 D NMR according to the peak intensity change of H-5 from IdoA and GlcA residues in the <sup>1</sup>H NMR spectrum, which is nondestructive of sample and also serves the kinetic property evaluation of immobilized C5-epi.</p></div>\",\"PeriodicalId\":1,\"journal\":{\"name\":\"Accounts of Chemical Research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":16.4000,\"publicationDate\":\"2020-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1080/07328303.2021.1903026\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Accounts of Chemical Research\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://www.sciencedirect.com/org/science/article/pii/S0732830322001379\",\"RegionNum\":1,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Accounts of Chemical Research","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/org/science/article/pii/S0732830322001379","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 1
摘要
d -葡萄糖醛酸c5 - epimase (C5-epi)是制备生物工程肝素的关键酶，可将d -葡萄糖醛酸转化为l -伊杜醛酸，为生产更安全的肝素提供了另一种方法。本研究提供了一种基于质子核磁共振(NMR)分析的方法来测定溴化氰固定c5 -外聚体的活性。该方法利用1H NMR光谱中IdoA和GlcA残基H-5的峰强度变化，可以实时监测C5-epi催化的反应，对样品无破坏性，也可用于评价固定化C5-epi的动力学性质。
1H NMR analysis of perdeutero N-sulfoheparosan C5-epimerization: a direct way to measure the activity of immobilized C5-epimerase
D-Glucuronyl C5-epimerase (C5-epi), which converts D-glucuronic acid to L-iduronic acid, is a crucial enzyme in the preparation of bioengineered heparin, which provides an alternative way to produce safer heparins. Herein, we provided an efficient way to measure the activity of cyanogen bromide-immobilized C5-epi based on proton nuclear magnetic resonance (NMR) analysis of perdeutero N-sulfoheparosan C5-epimerization. In this method, reactions catalyzed by C5-epi could be monitored by real-time 1 D NMR according to the peak intensity change of H-5 from IdoA and GlcA residues in the 1H NMR spectrum, which is nondestructive of sample and also serves the kinetic property evaluation of immobilized C5-epi.
期刊介绍：
Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance.
Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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