title: Muropeptide rescue in Bacillus subtilis involves sequential hydrolysis by beta-N-acetylglucosaminidase and N-acetylmuramyl-L-alanine amidase.
abstract: We identified a pathway in Bacillus subtilis that is used for recovery of N-acetylglucosamine (GlcNAc)-N-acetylmuramic acid (MurNAc) peptides (muropeptides) derived from the peptidoglycan of the cell wall. This pathway is encoded by a cluster of six genes, the first three of which are orthologs of Escherichia coli genes involved in N-acetylmuramic acid dissimilation and encode a MurNAc-6-phosphate etherase (MurQ), a MurNAc-6-phosphate-specific transcriptional regulator (MurR), and a MurNAc-specific phosphotransferase system (MurP). Here we characterized two other genes of this cluster. The first gene was shown to encode a cell wall-associated beta-N-acetylglucosaminidase (NagZ, formerly YbbD) that cleaves the terminal nonreducing N-acetylglucosamine of muropeptides and also accepts chromogenic or fluorogenic beta-N-acetylglucosaminides. The second gene was shown to encode an amidase (AmiE, formerly YbbE) that hydrolyzes the N-acetylmuramyl-L-Ala bond of MurNAc peptides but not this bond of muropeptides. Hence, AmiE requires NagZ, and in conjunction these enzymes liberate MurNAc by sequential hydrolysis of muropeptides. NagZ expression was induced at late exponential phase, and it was 6-fold higher in stationary phase. NagZ is noncovalently associated with lysozyme-degradable particulate material and can be released from it with salt. A nagZ mutant accumulates muropeptides in the spent medium and displays a lytic phenotype in late stationary phase. The evidence for a muropeptide catabolic pathway presented here is the first evidence for cell wall recovery in a Gram-positive organism, and this pathway is distinct from the cell wall recycling pathway of E. coli and other Gram-negative bacteria.
authors.affiliation: Fachbereich Biologie, Molecular Microbiology, University of Constance, 78457 Constance, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Litzinger
authors.forename: Amanda
authors.initials: A
authors.lastname: Duckworth
authors.forename: Katja
authors.initials: K
authors.lastname: Nitzsche
authors.forename: Christian
authors.initials: C
authors.lastname: Risinger
authors.forename: Valentin
authors.initials: V
authors.lastname: Wittmann
authors.forename: Christoph
authors.initials: C
authors.lastname: Mayer
mesh_terms: D000619:Aminohydrolases / Q000235:genetics / Q000378:metabolism*; D001412:Bacillus subtilis / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000502:physiology; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D010457:Peptidoglycan / Q000235:genetics / Q000378:metabolism; D048828:beta-N-Acetyl-Galactosaminidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan; D048828:beta-N-Acetyl-Galactosaminidase; D000619:Aminohydrolases
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