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Domain of Unknown Function 3380 (DUF3380) confers potent N-acetylmuramidase activity to a Salmonella phage endolysin

(2016)
Author
Organization
Abstract
Bacteriophage-encoded endolysins are highly diverse enzymes that cleave the bacterial peptidoglycan layer at the end of the lytic infection cycle to allow the viral progeny release. The specific activity and structure of these proteins have boosted their study as new antimicrobials against pathogens including multidrug resistant bacteria as well as the development of new biotechnological tools for bacterial diagnostics and detection, among others. Despite the wealth of applications relying on the use of endolysin, little is known about the enzymatic properties of these enzymes, especially in case of endolysins of bacteriophages infecting Gram-negative species. Only a limited number of studies have analyzed the peptidoglycan bond cleaved by endolysins and most annotations of enzymatic specificity only rely on sequence similarity. As a result, available databases contain inaccurate descriptions of biochemical specificities. In this study, we report the functional and biochemical characterization of the modular Salmonella phage endolysin Gp110 which comprises an uncharacterized Domain of Unknown Function (DUF3380; pfam11860) in its C-terminus and shows the highest specific activity (34,240 U/µM) compared to fourteen previously characterized endolysins active against peptidoglycan from Gram-negative bacteria (corresponding to a 1.7- to 364-fold higher activity). This endolysin showed an optimal enzymatic activity at pH 8 and an elevated thermal resistance. Reversed phase-HPLC analysis coupled to mass spectrometry showed that DUF3380 has N-acetylmuramidase (lysozyme) activity cleaving the β-(1,4) glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine residues. Gp110 is active against directly cross-linked peptidoglycan with various peptide stem compositions, making it an attractive enzyme to develop novel antimicrobial agents.

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Chicago
Rodríguez-Rubio, Lorena, Hans Gerstmans, Simon Thorpe, Stéphane Mesnage, Rob Lavigne, and Yves Briers. 2016. “Domain of Unknown Function 3380 (DUF3380) Confers Potent  N-acetylmuramidase Activity to a Salmonella Phage Endolysin.” In .
APA
Rodríguez-Rubio, L., Gerstmans, H., Thorpe, S., Mesnage, S., Lavigne, R., & Briers, Y. (2016). Domain of Unknown Function 3380 (DUF3380) confers potent  N-acetylmuramidase activity to a Salmonella phage endolysin. Presented at the EMBO Workshop Viruses of Microbes 2016.
Vancouver
1.
Rodríguez-Rubio L, Gerstmans H, Thorpe S, Mesnage S, Lavigne R, Briers Y. Domain of Unknown Function 3380 (DUF3380) confers potent  N-acetylmuramidase activity to a Salmonella phage endolysin. 2016.
MLA
Rodríguez-Rubio, Lorena et al. “Domain of Unknown Function 3380 (DUF3380) Confers Potent  N-acetylmuramidase Activity to a Salmonella Phage Endolysin.” 2016. Print.
@inproceedings{8623528,
  abstract     = {Bacteriophage-encoded endolysins are highly diverse enzymes that cleave the bacterial peptidoglycan layer at the end of the lytic infection cycle to allow the viral progeny release. The specific activity and structure of these proteins have boosted their study as new antimicrobials against pathogens including multidrug resistant bacteria as well as the development of new biotechnological tools for bacterial diagnostics and detection, among others. Despite the wealth of applications relying on the use of endolysin, little is known about the enzymatic properties of these enzymes, especially in case of endolysins of bacteriophages infecting Gram-negative species. Only a limited number of studies have analyzed the peptidoglycan bond cleaved by endolysins and most annotations of enzymatic specificity only rely on sequence similarity. As a result, available databases contain inaccurate descriptions of biochemical specificities.
In this study, we report the functional and biochemical characterization of the modular Salmonella phage endolysin Gp110 which comprises an uncharacterized Domain of Unknown Function (DUF3380; pfam11860) in its C-terminus and shows the highest specific activity (34,240 U/µM) compared to fourteen previously characterized endolysins active against peptidoglycan from Gram-negative bacteria (corresponding to a 1.7- to 364-fold higher activity). This endolysin showed an optimal enzymatic activity at pH 8 and an elevated thermal resistance. Reversed phase-HPLC analysis coupled to mass spectrometry showed that DUF3380 has N-acetylmuramidase (lysozyme) activity cleaving the β-(1,4) glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine residues. Gp110 is active against directly cross-linked peptidoglycan with various peptide stem compositions, making it an attractive enzyme to develop novel antimicrobial agents.},
  author       = {Rodríguez-Rubio, Lorena and Gerstmans, Hans and Thorpe, Simon and Mesnage, Stéphane and Lavigne, Rob and Briers, Yves},
  location     = {Liverpool},
  title        = {Domain of Unknown Function 3380 (DUF3380) confers potent  N-acetylmuramidase activity to a Salmonella phage endolysin},
  year         = {2016},
}