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Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples
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Lytic enzyme discovery through multigenomic sequence analysis in Clostridium perfringens.

Jonathan E Schmitz1, Maria Cristina Ossiprandi, Kareem R Rumah

  • 1Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, NY 10065, USA. jschmitz@rockefeller.edu

Applied Microbiology and Biotechnology
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Summary

Phage lysins, enzymes that break down bacteria, show promise as new antibiotics. A newly studied lysin, PlyCM, effectively targets the pathogen Clostridium perfringens, highlighting genomic data

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Area of Science:

  • Microbiology
  • Biotechnology
  • Genomics

Background:

  • Phage lytic enzymes (lysins) are proteins that degrade bacterial cell walls.
  • Lysins are recognized for their potential as novel anti-infective agents against Gram-positive bacteria.
  • Advances in DNA sequencing have accelerated the discovery of new lysin-encoding genes in bacterial genomes.

Purpose of the Study:

  • To investigate the diversity of lysins in the pathogen Clostridium perfringens.
  • To characterize a novel muramidase, PlyCM, from C. perfringens strain ATCC 13124 for its potential as an enzybiotic.

Main Methods:

  • Computational mining of nine C. perfringens genomes to identify prophage lysins and lysin-like open reading frames (ORFs).
  • Recombinant expression and purification of the selected muramidase, PlyCM.
  • In vitro biochemical characterization of PlyCM, including pH/salt dependence and temperature stability, and testing its activity against various bacterial strains.

Main Results:

  • Several dozen lysin-encoding ORFs were identified across the nine C. perfringens genomes.
  • Purified PlyCM demonstrated potent lytic activity at low concentrations (μg/ml).
  • PlyCM was highly specific, showing activity against 23 of 24 tested C. perfringens strains with minimal activity against other species.

Conclusions:

  • PlyCM exhibits significant potential as a targeted enzybiotic agent against Clostridium perfringens.
  • Expanding genomic databases are valuable resources for discovering novel biotechnologically relevant proteins like PlyCM.
  • Phage lysins represent a promising avenue for developing new strategies to combat bacterial infections.