Bacteriophage-derived enzyme that depolymerizes the alginic acid capsule associated with cystic fibrosis isolates of Pseudomonas aeruginosa

J Appl Microbiol. 2010 Feb;108(2):695-702. doi: 10.1111/j.1365-2672.2009.04469.x. Epub 2009 Jul 13.

Abstract

Aims: To identify enzymes associated with bacteriophages infecting cystic fibrosis (CF) strains of Pseudomonas aeruginosa that are able to degrade extracellular alginic acids elaborated by the host bacterium.

Methods and results: Plaques produced by 21 Ps. aeruginosa-specific phages were screened for the presence of haloes, an indicator of capsule hydrolytic activity. Four phages produced haloed plaques, and one (PT-6) was investigated further. PT-6 was shown by electron microscopy to belong to Podoviridae family C1, to reduce the viscosity of four alginate preparations using a rolling ball viscometer and to release uronic acid-containing fragments from the polymers, as judged by spectrophotometry and thin layer chromatography. The alginase was partially purified by gel filtration chromatography and shown to be a 37 kDa polypeptide.

Conclusions: Infection of CF strains of Ps. aeruginosa by phage PT-6 involves hydrolysis of the exopolysaccharide secreted by the host.

Significance and impact of the study: The alginase produced by PT-6 has the potential to increase the well-being of CF suffers by improving the surface properties of sputum, accelerating phagocytic uptake of bacteria and perturbing bacterial growth in biofilms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alginates / metabolism*
  • Bacterial Capsules / metabolism*
  • Bacteriophages / enzymology*
  • Biofilms
  • Cystic Fibrosis / microbiology*
  • Glucuronic Acid / metabolism
  • Hexuronic Acids / metabolism
  • Pseudomonas Infections / microbiology*
  • Pseudomonas aeruginosa / virology*
  • Viral Plaque Assay

Substances

  • Alginates
  • Hexuronic Acids
  • Glucuronic Acid