A nonenzymatic method for cleaving polysaccharides to yield oligosaccharides for structural analysis

• Published in: Nature communications Vol. 11; no. 1; pp. 3963 - 12
• Main Authors: Amicucci, Matthew J., Nandita, Eshani, Galermo, Ace G., Castillo, Juan Jose, Chen, Siyu, Park, Dayoung, Smilowitz, Jennifer T., German, J. Bruce, Mills, David A., Lebrilla, Carlito B.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 07.08.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 140/58; 631/1647/296; 631/45/1268; 631/45/72/1205; Bacteria - metabolism; Biomolecules; Digestion; Equivalence; Fingerprinting; Food; Food composition; Galactose - analogs & derivatives; Genomic analysis; Glucans - chemistry; Glucans - metabolism; Humanities and Social Sciences; Humans; Infant; Ions; Liquid chromatography; Mannans - chemistry; Mannans - metabolism; Mass spectrometry; Mass spectroscopy; multidisciplinary; Oligosaccharides; Oligosaccharides - chemistry; Oxidation-Reduction; Peptides; Polymerization; Polynucleotides; Polysaccharides; Polysaccharides - metabolism; Production methods; Proteins; Proteomics; Saccharides; Science; Science (multidisciplinary); Scientific imaging; Spectroscopy; Structural analysis; Time Factors; Trypsin; Workflow; Xylans - chemistry; Xylans - metabolism
• ISSN: 2041-1723; 2041-1723
• 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. 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.