Related Experiment Videos
Bacteriophage and associated polysaccharide depolymerases--novel tools for study of bacterial biofilms
K A Hughes1, I W Sutherland, J Clark
1Institute of Cell and Molecular Biology, University of Edinburgh, UK. K.Hughes@BAS.ac.uk
Journal of Applied Microbiology
|September 29, 1998
Summary
Lytic bacteriophages targeting Gram-negative biofilm bacteria were isolated and found to produce enzymes that degrade bacterial exopolysaccharides. These bacteriophages and their enzymes are valuable tools for studying biofilms and offer potential applications in biofilm control.
Area of Science:
- Microbiology
- Virology
- Biochemistry
Background:
- Bacteriophages are viruses that infect bacteria and are widespread in the environment.
- Biofilms are complex communities of microorganisms that can cause significant problems in industrial and medical settings.
- Exopolysaccharide (EPS) production is a key feature of many biofilm-forming bacteria, contributing to biofilm structure and protection.
Purpose of the Study:
- To isolate and characterize lytic bacteriophages targeting Gram-negative biofilm bacteria.
- To investigate the enzymatic activity of these bacteriophages, particularly their ability to degrade exopolysaccharides.
- To explore the potential applications of bacteriophages and their associated enzymes in biofilm research.
Main Methods:
- Isolation of lytic bacteriophages from sewage targeting specific bacterial strains (Pseudomonad 1.15 and Enterobacter agglomerans).
- Characterization of bacteriophage plaque morphology and enzymatic activity on solid media.
- Partial purification of phage-associated polysaccharase enzymes and analysis of polysaccharide digests.
- Enzymatic assays to determine the mode of action (endo-glycanohydrolase activity).
Main Results:
- Widespread occurrence of lytic bacteriophages against Gram-negative biofilm bacteria was confirmed.
- Isolated bacteriophages produced large plaques with halos, indicating polysaccharide depolymerase activity.
- Phage-associated enzymes degraded bacterial exopolysaccharides into presumed repeat units or oligomers.
- Enzymatic activity was associated with both phage particles and bacterial lysates.
Conclusions:
- Bacteriophages and their associated enzymes are effective tools for degrading bacterial exopolysaccharides.
- These phage-derived enzymes act as endo-glycanohydrolases, specifically targeting EPS.
- The isolated bacteriophages and their enzymes offer highly specific tools for studying biofilms and have potential applications in biofilm management.