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New-Generation Antibacterial Agent-Cellulose-Binding Thermostable TP84_Endolysin.

Małgorzata Ponikowska1,2, Joanna Żebrowska1, Piotr M Skowron1

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Researchers engineered a fusion protein combining a cellulose-binding domain with thermostable endolysin TP84_28. This novel antibacterial agent targets biofilms and resistant bacteria, enhancing efficacy at specific sites.

Keywords:
TP-84antimicrobialbacteriophagecellulose-binding domainendolysinthermophage

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Area of Science:

  • Biotechnology and Molecular Biology
  • Antimicrobial Resistance Research
  • Enzyme Engineering

Background:

  • Increasing antibiotic resistance necessitates novel antibacterial strategies.
  • Bacterial biofilms pose significant challenges in clinical and industrial settings.
  • Efficient delivery of antibacterial agents to target sites remains a critical issue.

Purpose of the Study:

  • To develop a targeted antibacterial agent by fusing a cellulose-binding domain (CBD) to a thermostable endolysin.
  • To evaluate the antibacterial activity, thermostability, and biofilm removal capabilities of the fusion protein.
  • To assess the potential of CBD-endolysin fusion proteins for localized antibacterial applications.

Main Methods:

  • Cloning of the endolysin TP84_28 gene with a CBD into an expression vector.
  • Biosynthesis and purification of the Cellulose-Binding Thermostable TP84_Endolysin (CBD_TP84_28_His) in *Escherichia coli*.
  • Assays for enzymatic activity (turbidity reduction, spot assay), biofilm removal, thermostability, and cellulose-binding properties.

Main Results:

  • CBD_TP84_28_His demonstrated effective lytic activity against thermophilic bacteria and significant biofilm reduction.
  • The fusion protein exhibited high thermostability, retaining activity up to 73 °C.
  • Cellulose-binding properties were confirmed, enabling targeted delivery to cellulose-based materials.

Conclusions:

  • Fusion of CBDs to thermostable endolysins offers a promising strategy for developing targeted antibacterial agents.
  • CBD_TP84_28_His shows potential for applications in wound dressings, medical devices, and food packaging.
  • This approach could lead to novel recombinant proteins to combat antibiotic resistance and biofilm infections.