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Targeting Biofilms: Enzymatic Phage Engineering for Dispersal

This study explores using engineered enzymatic bacteriophages to disperse biofilms. Biofilms are known for their antibiotic resistance and pose challenges in various industries. The research focuses on degrading the essential adhesin in bacterial biofilms using engineered phages with specific genes. The results show significant reductions in biofilm cell counts, indicating promising potential for this approach. The study outlines a practical application of phage engineering without altering replication, highlighting the effectiveness of the method.

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Targeting Biofilms: Enzymatic Phage Engineering for Dispersal

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  1. Dispersing biofilms with engineeredenzymatic bacteriophage Doug Tischer

  2. The Problem with Biofilms • Increases resistant to antibiotics • Pose a problem for cleaning medical devices, water pipe, food contamination from industrial devices etc • EPS (Extracellular Polymeric Substance) • Polysaccharides • Proteins • Nucleic Acids • Lipids

  3. The Idea • Adhesin polymeric β-1,6-N-acetyl-D-glucosamine essential for bacterial adhesion to biofilm • DspB degrades polymeric β -1,6-N-acetyl-D-glucosamine • T3 gene 1.2 allows replication in F-plasmid-containing E. coli Figure taken from Lu, 2007

  4. Construction • T7wt • T7DspB • T7control • Φ10 promotor expresses only during infection • 10B is a capsid protein • S-tag easily detected in western blot Figure taken from Lu, 2007

  5. Results Crystal Violet Staining CFU Counts PFU Counts • No replication (no T3 gene 1.2) Inoculation • T3 gene 1.2 greatly enhances phage efficacy in F-plasmid containing E. coli • DspB enhances plaque destruction • Engineered phage is replicating • PFU = Plaque forming unit Figure taken from Lu, 2007

  6. Results cont. Treated Untreated • 4.5 orders of magnitude drop = 99.997% drop in cell count • Scanning Electron Microscope of cell plaques • Significant disruption of plaques Figure taken from Lu, 2007

  7. My thoughts Liked Disliked • Very practical application • Showed phages can be engineered with several genes of interest without hindering replication • Successful proof of principle • Relatively simple construction • No new behavioral tricks • Only protein expression • No use of premade parts

  8. References • Lu TK, Collins JJ (2007) PNAS 104:11197–11202 • Mark McCormick and Robert Mierendorf. “S•Tag: A Multipurpose Fusion Peptide for Recombinant Proteins.” Novagen Inc.

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