TY - JOUR T1 - Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown JF - bioRxiv DO - 10.1101/835843 SP - 835843 AU - Lucy I. Crouch AU - Marcelo V. Liberato AU - Paulina A. Urbanowicz AU - Arnaud Baslé AU - Christopher A. Lamb AU - Christopher J. Stewart AU - Katie Cooke AU - Mary Doona AU - Stephanie Needham AU - Richard R. Brady AU - Janet E. Berrington AU - Katarina Madunic AU - Manfred Wuhrer AU - Peter Chater AU - Jeffery P. Pearson AU - Robert Glowacki AU - Eric C. Martens AU - Fuming Zhang AU - Robert J. Linhardt AU - Daniel I. R. Spencer AU - David N. Bolam Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/11/09/835843.abstract N2 - The human gut microbiota (HGM) are closely associated with health, development and disease. The thick intestinal mucus layer, especially in the colon, is the key barrier between the contents of the lumen and the epithelial cells, providing protection against infiltration by the microbiota as well potential pathogens. The upper layer of the colonic mucus is a niche for a subset of the microbiota which utilise the mucin glycoproteins as a nutrient source and mucin grazing by the microbiota appears to play a key role in maintaining barrier function as well as community stability. Despite the importance of mucin breakdown for gut health, the mechanisms by which gut bacteria access this complex glycoprotein are not well understood. The current model for mucin degradation involves exclusively exo-acting glycosidases that sequentially trim monosaccharides from the termini of the glycan chains to eventually allow access to the mucin peptide backbone by proteases. However, this model is in direct contrast to the Sus paradigm of glycan breakdown used by the Bacteroidetes which involves extracellular cleavage of glycans by surface located endo-acting enzymes prior to import of the oligosaccharide products. Here we describe the discovery and characterisation of endo-acting family 16 glycoside hydrolases (GH16s) from prominent mucin degrading gut bacteria that specifically target the oligosaccharide side chains of intestinal mucins from both animals and humans. These endo-acting O-glycanases display β1,4-glactosidase activity and in several cases are surface located indicating they are involved in the initial step in mucin breakdown. The data suggest a new paradigm for mucin breakdown by the microbiota and the endo-mucinases provide a potential tool to explore changes that occur in mucin structure in intestinal disorders such as inflammatory bowel disease and colon cancer. ER -