T4 lysozyme fused with cellulose-binding module for antimicrobial cellulosic wound dressing materials

Area of Science:

• Biotechnology
• Materials Science
• Microbiology

Background:

• Bacterial infections pose significant challenges in wound care, necessitating advanced antimicrobial strategies.
• Endolysins, such as T4 lysozyme (T4Lyz), represent a promising new class of antimicrobials with diverse applications.
• The development of effective antimicrobial wound dressings is crucial for managing wound infections and addressing antibiotic resistance.

Purpose of the Study:

• To produce and evaluate T4 lysozyme (T4Lyz) fused with a Cellulose Binding Module (CBM) for immobilization onto cellulosic wound dressings.
• To assess the antimicrobial activity and stability of the T4Lyz-CBM fusion protein, both free and immobilized.
• To present a novel strategy for creating advanced antimicrobial wound dressing materials.

Main Methods:

• Genes for T4Lyz-CBM and T4Lyz were cloned and expressed in Escherichia coli, followed by purification using cation exchange chromatography.
• The lytic and bactericidal activities of T4Lyz-CBM were determined using viable count plating assays against various bacteria.
• Immobilization capacity of T4Lyz-CBM onto cellulosic wound dressing gauze was quantified, and binding of unmodified T4Lyz was assessed.

Main Results:

• The T4Lyz-CBM fusion protein demonstrated significant bactericidal activity against Gram-positive and Gram-negative bacteria.
• Immobilization of T4Lyz-CBM onto cellulosic gauze was achieved with a capacity of 5.5 μg/mg, while unmodified T4Lyz showed no binding.
• The immobilized T4Lyz-CBM retained its bactericidal activity and exhibited enhanced stability compared to the free enzyme, particularly in dry storage or with polyol stabilizers.

Conclusions:

• Fusion of T4 lysozyme with a Cellulose Binding Module enables facile and irreversible binding to cellulosic wound dressings, preserving antimicrobial activity.
• This strategy offers a viable method for developing innovative antimicrobial wound dressing materials.
• The approach holds potential for application with other antimicrobial enzymes and peptides to combat antibiotic resistance.