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

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. Plaques produced by 21 Ps. aeruginosa-specific phages were screened for the presence of haloes, a...

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Bibliographic Details
Published in:Journal of applied microbiology 2010-02, Vol.108 (2), p.695-702
Main Authors: Glonti, T, Chanishvili, N, Taylor, P.W
Format: Article
Language:English
Subjects:
alginate lyase
alginates
Alginates - metabolism
alginic acid
Bacteria
Bacterial Capsules - metabolism
bacteriophage
Bacteriophages - enzymology
Biofilms
Biological and medical sciences
Cystic Fibrosis - microbiology
exopolysaccharide depolymerase
Fundamental and applied biological sciences. Psychology
Glucuronic Acid - metabolism
Hexuronic Acids - metabolism
Microbiology
Podoviridae
Pseudomonas aeruginosa
Pseudomonas aeruginosa - virology
Pseudomonas Infections - microbiology
Viral Plaque Assay
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Summary: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. 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. Infection of CF strains of Ps. aeruginosa by phage PT-6 involves hydrolysis of the exopolysaccharide secreted by the host. 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.
ISSN:1364-5072
1365-2672
DOI:10.1111/j.1365-2672.2009.04469.x