Agricultural and Environmental Chemistry Graduate Group, University of California, Davis, CA, USA.
Department of Chemistry, University of California, Davis, CA, USA.
Nat Commun. 2020 Aug 7;11(1):3963. doi: 10.1038/s41467-020-17778-1.
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.
多糖是自然界中最丰富的生物分子，但就其化学结构和生物功能而言，它们是了解最少的。多糖不能简单地进行测序，因为它们通常具有高度支化且缺乏均匀的结构。此外，由于质量 spectrometry 技术无法直接分析大型聚合结构，因此这一问题已在核酸和蛋白质中得到解决。尽管限制性内切酶可用于高级基因组分析，胰蛋白酶可用于高级蛋白质组学分析，但目前还没有用于通用多糖消化的等效酶。我们描述了一种从多糖中产生寡糖的化学方法的开发和应用。释放的寡糖通过具有高灵敏度、准确性和通量的先进液相色谱-质谱（LC-MS）方法进行表征。该技术首先用于通过寡糖指纹图谱鉴定多糖。接下来，确定食物和粪便中的多糖组成，进一步说明了该技术在食品和临床研究中的实用性。
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