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

IF 1.2 4区化学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Journal of Carbohydrate Chemistry Pub Date : 2020-01-01 DOI:10.1080/07328303.2021.1903026

Meng Qiao , Ruoyu Jiao , Xing Zhang , Zhengzhang Wu , Baoxing Shen , Bingzhi Li

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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。

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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.

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来源期刊

Journal of Carbohydrate Chemistry 化学-生化与分子生物学

CiteScore

2.10

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： The Journal of Carbohydrate Chemistry serves as an international forum for research advances involving the chemistry and biology of carbohydrates. The following aspects are considered to fall within the scope of this journal: -novel synthetic methods involving carbohydrates, oligosaccharides, and glycoconjugates- the use of chemical methods to address aspects of glycobiology- spectroscopic and crystallographic structure studies of carbohydrates- computational and molecular modeling studies- physicochemical studies involving carbohydrates and the chemistry and biochemistry of carbohydrate polymers.