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Publication Abstract from PubMed

beta-N-acetylhexosaminidases (EC3.2.1.52), which belong to the glycosyl hydrolase family GH20, are important enzymes for oligosaccharides modification. Numerous microbial beta-N-acetylhexosaminidases have been investigated for applications in biology, biomedicine and biotechnology. Akkermansia muciniphila is an anaerobic intestinal commensal bacterium which possesses specific beta-N-acetylhexosaminidases for gut mucosal layer colonization and mucin degradation. In this study, we assessed the in vitro mucin glycan cleavage activity of the A. muciniphila beta-N-acetylhexosaminidase Am2136 and demonstrated its ability that hydrolyzing the beta-linkages joining N-acetylglucosamine to a wide variety of aglycone residues, which indicated that Am2136 may be a generalist beta-N-acetylhexosaminidase. Structural and enzyme activity assay experiments allowed us to probe the essential function of the inter-domain interactions in beta23-beta33. Importantly, we revealed that the hydrolysis activity of Am2136 was enhanced by nucleotides. We further speculated that this activation mechanism might be associated with the conformational motions between domain III and IV. To our knowledge, this is the first report of nucleotide effector regulated beta-N-acetylhexosaminidase, to reveal its novel biological functions. These findings contribute to understanding the distinct properties within the GH20 family and lay a certain foundation to develop controllable glycan hydrolyzing catalysts.Abbreviations: OD600 - optical cell densities at 600 nm; LB - Luria-Bertani; IPTG - isopropyl beta-D-1-thiogalactopyranoside; PMSF - phenylmethanesulfonyl fluoride; rmsd - root mean square deviation; GlcNAc - N-acetyl-beta-D-glucosamine; GalNAc - N-acetyl-beta-D-galactosamine; Gal - galactose.

Nucleotide binding as an allosteric regulatory mechanism for Akkermansia muciniphila beta-N-acetylhexosaminidase Am2136.,Li CC, Yi H, Wang YM, Tang XY, Zhu YB, Song YJ, Zhao NL, Huang Q, Mou XY, Luo GH, Liu TG, Yang GL, Zeng YJ, Wang LJ, Tang H, Fan G, Bao R Gut Microbes. 2022 Jan-Dec;14(1):2143221. doi: 10.1080/19490976.2022.2143221. PMID:36394293^[2]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.