Bacterial Enzyme Assay for Mucin Glycan Degradation

Bacterial sialidase and sulfoglycosidase may act on the acidic modifications of mucin O-glycans, producing sialic acid and 6-sulfated N-acetylglucosamine, respectively. Assays for these enzymes, using mucin as a substrate, are enabled by the detection and/or quantification of the free monosaccharides that are released by these enzymes. This chapter describes enzyme reactions with mucin, detection by thin-layer chromatography of sialic acid, and quantification of 6-sulfated N-acetylglucosamine by liquid chromatography-tandem mass spectrometry.