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Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins
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Characterization of a phiBP endolysin encoded by the Paenibacillus polymyxa CCM 7400 phage

Jana Ugorcakova1, Livia Medzova1, Barbora Solteszova1

  • 1Department of Genomics and Biotechnology, Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 845 51 Bratislava, Slovakia.

FEMS Microbiology Letters
|June 19, 2015
PubMed

Insights

The Paenibacillus polymyxa phage phiBP endolysin requires its cell wall-binding domain for full lytic activity against Paenibacillus bacteria. This binding domain is crucial for the endolysin

Area of Science:

  • Bacteriophage biology
  • Enzymology
  • Microbial genetics

Background:

  • Endolysins are phage-encoded enzymes that degrade bacterial cell walls for progeny release.
  • The Paenibacillus polymyxa phage phiBP endolysin (gp1.2) possesses a modular structure with catalytic and cell wall-binding domains.
  • Understanding endolysin domain function is key to developing novel antibacterial agents.

Purpose of the Study:

  • To investigate the functional roles of the catalytic and cell wall-binding domains of phiBP endolysin.
  • To determine the necessity of the cell wall-binding domain for endolysin activity and substrate specificity.
  • To confirm the hydrolytic capability of phiBP endolysin on various bacterial cell walls.

Main Methods:

  • Cloning and recombinant expression of full-length phiBP endolysin and its domain fragments in Escherichia coli.
  • Purification of recombinant proteins using affinity chromatography.
  • Assays to evaluate lytic activity against cell wall substrates from Paenibacillus, Bacillus, Corynebacterium, and E. coli.
  • Specific binding assays using the cell wall-binding domain fused to green fluorescent protein.

Main Results:

  • The full-length phiBP endolysin exhibited lytic activity, while the isolated catalytic domain showed reduced activity.
  • The cell wall-binding domain was essential for the full lytic activity of the phiBP endolysin.
  • The binding domain demonstrated specific binding to the cell surface of Paenibacillus polymyxa CCM 7400 and other Paenibacillus strains.

Conclusions:

  • The cell wall-binding domain is indispensable for the optimal hydrolytic function of phiBP endolysin.
  • phiBP endolysin specifically targets and degrades the cell walls of Paenibacillus species.
  • These findings highlight the importance of domain cooperation for endolysin efficacy.
Keywords:
bacteriophagecell lysisendolysinfluorescence microscopy

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