The exo-β-N-acetylmuramidase NamZ from Bacillus subtilis is the founding member of a family of exo-lytic peptidoglycan hexosaminidases

Muller, Maraike; Calvert, Matthew; Hottmann, Isabel; Kluj, Robert Maria; Teufel, Tim; Balbuchta, Katja; Engelbrecht, Alicia; Selim, Khaled A.; Xu, Qingping; Borisova, Marina; Titz, Alexander; Mayer, Christoph

doi:10.1016/j.jbc.2021.100519

Title: The exo-β-N-acetylmuramidase NamZ from Bacillus subtilis is the founding member of a family of exo-lytic peptidoglycan hexosaminidases

Journal Article · Fri Mar 05 00:00:00 EST 2021 · Journal of Biological Chemistry

DOI:https://doi.org/10.1016/j.jbc.2021.100519· OSTI ID:1819671

Muller, Maraike ^[1]; Calvert, Matthew ^[2]; Hottmann, Isabel ^[1]; Kluj, Robert Maria ^[1]; Teufel, Tim ^[1]; Balbuchta, Katja ^[1]; Engelbrecht, Alicia ^[1];

^[3]; Xu, Qingping ^[4];

^[1];

^[2];

^[5]

Univ. of Tubingen (Germany)
Helmholtz Centre for Infection Research (HZI), Saarbrucken (Germany); Deutsches Zentrum fur Infektionsforschung (DZIF), Standort Hannover-Braunschweig (Germany); Saarland Univ., Saarbrucken (Germany)
Univ. of Tubingen (Germany); National Research Center, Giza (Egypt)
Argonne National Lab. (ANL), Lemont, IL (United States)
Univ. of Tubingen (Germany); Univ. of Auckland (New Zealand)

Endo-β-N-acetylmuramidases, commonly known as lysozymes, are well-characterized antimicrobial enzymes that catalyze an endo-lytic cleavage of peptidoglycan; i.e., they hydrolyze the β-1,4-glycosidic bonds connecting N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc). In contrast, little is known about exo-β-N-acetylmuramidases, which catalyze an exo-lytic cleavage of β-1,4-MurNAc entities from the non-reducing ends of peptidoglycan chains. Such an enzyme was identified earlier in the bacterium Bacillus subtilis, but the corresponding gene has remained unknown so far. We now report that ybbC of B. subtilis, renamed namZ, encodes the reported exo-β-N-acetylmuramidase. A ΔnamZ mutant accumulated specific cell wall fragments and showed growth defects under starvation conditions, indicating a role of NamZ in cell wall turnover and recycling. Recombinant NamZ protein specifically hydrolyzed the artificial substrate para-nitrophenyl β-MurNAc and the peptidoglycan-derived disaccharide MurNAc-β-1,4-GlcNAc. Together with the exo-β-N-acetylglucosaminidase NagZ and the exo-muramoyl-L-alanine amidase AmiE, NamZ degraded intact peptidoglycan by sequential hydrolysis from the non-reducing ends. A structure model of NamZ, built on the basis of two crystal structures of putative orthologs from Bacteroides fragilis, revealed a two-domain structure including a Rossmann-fold-like domain that constitutes a unique glycosidase fold. Thus, NamZ, a member of the DUF1343 protein family of unknown function, is now classified as the founding member of a new family of glycosidases (CAZy GH171). NamZ-like peptidoglycan hexosaminidases are mainly present in the phylum Bacteroidetes and less frequently found in individual genomes within Firmicutes (Bacilli, Clostridia), Actinobacteria, and γ-proteobacteria.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Organization:: National Institutes of Health (NIH); German Research Foundation (DFG)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1819671

Journal Information:: Journal of Biological Chemistry, Vol. 296; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: English

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
cell wall recycling
N-acetylglucosaminidase
N-acetylmuramidase
N-acetylmuramoyl amidase
Rossmann-fold
exo-lytic glycosidase
lysozyme
peptidoglycan hydrolase

Title: The exo-β-N-acetylmuramidase NamZ from Bacillus subtilis is the founding member of a family of exo-lytic peptidoglycan hexosaminidases

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