Permethylation as a strategy for high-molecular-weight polysaccharide structure analysis by nuclear magnetic resonance—Case study of Xylella fastidiosa extracellular polysaccharide

IF 1.9 3区 化学 Q3 CHEMISTRY, MULTIDISCIPLINARY
Ikenna E. Ndukwe, Ian Black, Claudia A. Castro, Jiri Vlach, Christian Heiss, Caroline Roper, Parastoo Azadi
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引用次数: 0

Abstract

Current practices for structural analysis of extremely large-molecular-weight polysaccharides via solution-state nuclear magnetic resonance (NMR) spectroscopy incorporate partial depolymerization protocols that enable polysaccharide solubilization in suitable solvents. Non-specific depolymerization techniques utilized for glycosidic bond cleavage, such as chemical degradation or ultrasonication, potentially generate structural fragments that can complicate complete and accurate characterization of polysaccharide structures. Utilization of appropriate enzymes for polysaccharide degradation, on the other hand, requires prior structural knowledge and optimal enzyme activity conditions that are not available to an analyst working with novel or unknown compounds. Herein, we describe an application of a permethylation strategy that allows the complete dissolution of intact polysaccharides for NMR structural characterization. This approach is utilized for NMR analysis of Xylella fastidiosa extracellular polysaccharide (EPS), which is essential for the virulence of the plant pathogen that affects multiple commercial crops and is responsible for multibillion dollar losses each year.

Abstract Image

Abstract Image

核磁共振分析高分子量多糖结构的过甲基化策略——以苛化木藻胞外多糖为例。
目前通过溶液态核磁共振(NMR)波谱法对特大分子量多糖进行结构分析的实践包括部分解聚方案,使多糖在合适的溶剂中溶解。用于糖苷键裂解的非特异性解聚技术,如化学降解或超声,可能会产生结构片段,使多糖结构的完整和准确表征复杂化。另一方面,利用适当的酶进行多糖降解,需要事先的结构知识和最佳的酶活性条件,这对于研究新的或未知化合物的分析人员来说是不可用的。在这里,我们描述了一个应用过甲基化策略,允许完整的多糖的核磁共振结构表征完全溶解。该方法被用于对苛化木藻胞外多糖(EPS)的NMR分析,EPS对影响多种经济作物的植物病原体的毒力至关重要,每年造成数十亿美元的损失。
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来源期刊
CiteScore
4.70
自引率
10.00%
发文量
99
审稿时长
1 months
期刊介绍: MRC is devoted to the rapid publication of papers which are concerned with the development of magnetic resonance techniques, or in which the application of such techniques plays a pivotal part. Contributions from scientists working in all areas of NMR, ESR and NQR are invited, and papers describing applications in all branches of chemistry, structural biology and materials chemistry are published. The journal is of particular interest not only to scientists working in academic research, but also those working in commercial organisations who need to keep up-to-date with the latest practical applications of magnetic resonance techniques.
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