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

被引：2

作者：

Ndukwe, Ikenna E. ^{[1
,4
]}

Black, Ian ^{[1
]}

Castro, Claudia A. ^{[2
]}

Vlach, Jiri ^{[1
]}

Heiss, Christian ^{[1
,3
]}

Roper, Caroline ^{[2
]}

Azadi, Parastoo ^{[1
]}

机构：

[1] Univ Georgia, Complex Carbohydrate Res Ctr, Athens, GA USA

[2] Univ Calif Riverside, Dept Microbiol & Plant Pathol, Riverside, CA USA

[3] Univ Georgia, Complex Carbohydrate Res Ctr, 315 Riverbend Rd, Athens, GA 30602 USA

[4] Amgen Inc, Pivotal Attribute Sci, Thousand Oaks, CA USA

来源：

MAGNETIC RESONANCE IN CHEMISTRY | 2024年 / 62卷 / 05期

基金：

美国国家卫生研究院;

关键词：

C-13; H-1; exopolysaccharide; high molecular weight; NMR; permethylation; solution-state NMR; Xylella fastidiosa; GLYCOSIDIC LINKAGES; SELECTIVE CLEAVAGE; EXOPOLYSACCHARIDE;

D O I：

10.1002/mrc.5413

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

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.

引用

页码：370 / 377

页数：8