1887

Abstract

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 strains were investigated. First, the genome sequence of CD034 and NRRL B-30929 was analysed for the presence of PGHs. Of 23 putative PGHs with different predicted hydrolytic specificities, the glycosyl hydrolase family 25 domain-containing homologues GH25B and GH25N from 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 strains CD034 and NRRL B-30929, and from GG, which served as a reference, revealed that GH25B and GH25N have -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 GH25B and GH25N to bind to cell walls . Moreover, similar secondary structures mainly composed of β-sheets and nearly identical thermal stabilities with values around 49 °C were found for the two -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 -acetylmuramidase from was characterized in detail for the first time.

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2014-08-01
2019-10-19
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