A nonenzymatic method for cleaving polysaccharides to yield oligosaccharides for structural analysis
A nonenzymatic method for cleaving polysaccharides to yield oligosaccharides for structural analysis
复制标题
DOI：
10.1038/s41467-020-17778-1
复制
发表时间：
2020-08-07
影响因子：
16.6
通讯作者：
Lebrilla, Carlito B.
中科院分区：
文献类型：
--
作者：
Amicucci, Matthew J.;Nandita, Eshani;Lebrilla, Carlito B.
多糖是自然界中最丰富的生物分子，但在其化学结构和生物功能方面了解得最少。多糖不能简单地测序，因为它们通常是高度分支的，缺乏统一的结构。此外，大的聚合物结构不能通过质谱技术直接分析，这是多核苷酸和蛋白质已经解决的问题。虽然限制性内切酶具有先进的基因组分析，胰蛋白酶具有先进的蛋白质组学分析，但还没有用于通用多糖消化的等效酶。本文介绍了一种从多糖中制备低聚糖的化学方法的发展和应用。释放的低聚糖通过具有高灵敏度、准确度和通量的先进液相色谱-质谱（LC-MS）方法进行表征。该技术首次用于通过寡糖指纹图谱鉴定多糖。接下来，测定食物和粪便中的多糖组成，进一步说明该技术在食品和临床研究中的实用性。虽然基于质谱的蛋白质组学在很大程度上依赖于将蛋白质消化成肽，但对于多糖分析没有等效的策略。在这里，作者开发了一种化学方法，将聚分解为寡糖，并提出了一种通过寡糖指纹识别多糖的工作流程。
Polysaccharides are the most abundant biomolecules in nature, but are the least understood in terms of their chemical structures and biological functions. Polysaccharides cannot be simply sequenced because they are often highly branched and lack a uniform structure. Furthermore, large polymeric structures cannot be directly analyzed by mass spectrometry techniques, a problem that has been solved for polynucleotides and proteins. While restriction enzymes have advanced genomic analysis, and trypsin has advanced proteomic analysis, there has been no equivalent enzyme for universal polysaccharide digestion. We describe the development and application of a chemical method for producing oligosaccharides from polysaccharides. The released oligosaccharides are characterized by advanced liquid chromatography-mass spectrometry (LC-MS) methods with high sensitivity, accuracy and throughput. The technique is first used to identify polysaccharides by oligosaccharide fingerprinting. Next, the polysaccharide compositions of food and feces are determined, further illustrating the utility of technique in food and clinical studies. While mass spectrometry-based proteomics largely relies on digesting proteins into peptides, there is no equivalent strategy for polysaccharide analysis. Here, the authors develop a chemical approach to break down poly- into oligosaccharides and present a workflow to identify polysaccharides by oligosaccharide fingerprinting.