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Gewählte Publikation:

Anzengruber, J; Courtin, P; Claes, IJ; Debreczeny, M; Hofbauer, S; Obinger, C; Chapot-Chartier, MP; Vanderleyden, J; Messner, P; Schäffer, C; .
(2014): Biochemical characterization of the major N-acetylmuramidase from Lactobacillus buchneri.
Microbiology. 2014; 160(Pt 8):1807-1819 FullText FullText_BOKU

Bacterial cell wall hydrolases are essential for peptidoglycan remodelling in regard to bacterial cell growth and division. In this study, peptidoglycan hydrolases (PGHs) of different Lactobacillus buchneri strains were investigated. First, the genome sequence of L. buchneri CD034 and L. buchneri NRRL B-30929 was analysed in silico for the presence of PGHs. Of 23 putative PGHs with different predicted hydrolytic specificities, the glycosyl hydrolase family 25 domain-containing homologues LbGH25B and LbGH25N from L. buchneri CD034 and NRRL B-30929, respectively, were selected and characterized in detail. Zymogram analysis confirmed hydrolysing activity on bacterial cell walls for both enzymes. Subsequent reversed-phase HPLC and MALDI-TOF MS analysis of the peptidoglycan breakdown products from L. buchneri strains CD034 and NRRL B-30929, and from Lactobacillus rhamnosus GG, which served as a reference, revealed that LbGH25B and LbGH25N have N-acetylmuramidase activity. Both enzymes were identified as cell wall-associated proteins by means of immunofluorescence microscopy and cellular fractionation, as well as by the ability of purified recombinant LbGH25B and LbGH25N to bind to L. buchneri cell walls in vitro. Moreover, similar secondary structures mainly composed of beta-sheets and nearly identical thermal stabilities with T-m values around 49 degrees C were found for the two N-acetylmuramidases by far-UV circular dichroism spectroscopy. The functional and structural data obtained are discussed and compared to related PGHs. In this study, a major N-acetylmuramidase from L. buchneri was characterized in detail for the first time.
Autor*innen der BOKU Wien:
Anzengruber Julia
Debreczeny Monika
Hofbauer Stefan
Messner Paul
Obinger Christian
Schäffer Christina
Find related publications in this database (using NML MeSH Indexing)
Bacterial Proteins/chemistry*;Bacterial Proteins/genetics;Bacterial Proteins/metabolism;Cell Wall/chemistry;Cell Wall/enzymology;Cell Wall/genetics;Enzyme Stability;Glycoside Hydrolases/chemistry*;Glycoside Hydrolases/genetics;Glycoside Hydrolases/metabolism;Lactobacillus/chemistry;Lactobacillus/enzymology*;Lactobacillus/genetics;Peptidoglycan/metabolism;Protein Structure, Secondary;Protein Transport;

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