Bacterial Enzyme Assay for Mucin Glycan Degradation

Toshihiko Katoh; Hisashi Ashida

doi:10.1007/978-1-0716-3670-1_28

Bacterial Enzyme Assay for Mucin Glycan Degradation

Methods Mol Biol. 2024:2763:337-344. doi: 10.1007/978-1-0716-3670-1_28.

Authors

Toshihiko Katoh¹, Hisashi Ashida²

Affiliations

¹ Graduate School of Biostudies, Kyoto University, Kyoto, Japan. tkatoh@lif.kyoto-u.ac.jp.
² Faculty of Biology-Oriented Science and Technology, Kindai University, Kinokawa, Wakayama, Japan.

PMID: 38347423
DOI: 10.1007/978-1-0716-3670-1_28

Abstract

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.

Keywords: Bifidobacterium bifidum; LC-MS/MS; Mucin; O-Glycan; Sialidase; Sulfoglycosidase; TLC.

MeSH terms

Acetylglucosamine / metabolism
Bacteria / metabolism
Carbohydrate Metabolism
Mucins* / metabolism
N-Acetylneuraminic Acid* / metabolism
Polysaccharides / metabolism
Sulfates

Substances

Mucins
N-Acetylneuraminic Acid
Acetylglucosamine
Polysaccharides
Sulfates