A Bacteriophage Holin-Derived Antimicrobial Peptide with Membrane-Disruptive Activity and Therapeutic Efficacy Against MDR Pseudomonas aeruginosa Burn Wound Infections

Introduction: Multidrug-resistant (MDR) Pseudomonas aeruginosa is a major cause of burn wound infections and represents a significant antimicrobial resistance challenge. Antimicrobial peptides (AMPs) derived from bacteriophage proteins offer a promising alternative to conventional antibiotics. This study aimed to identify and evaluate a novel bacteriophage holin-derived AMP against MDR P. aeruginosa.

Methods: An artificial intelligence-guided approach was employed to identify AMP, a 12-amino-acid cationic peptide derived from a Pseudomonas bacteriophage holin protein. Antibacterial, antibiofilm, membrane-disruption, stability, hemocompatibility, and cytotoxicity assays were performed. Therapeutic efficacy was further evaluated using a carbomer hydrogel formulation in a murine burn wound infection model caused by MDR P. aeruginosa.

Results: AMP exhibited potent antibacterial and antibiofilm activity against P. aeruginosa, achieving complete bacterial killing within 90 min. Mechanistic studies demonstrated concentration- and time-dependent membrane permeabilization and disruption. The peptide showed minimal cytotoxicity, high hemocompatibility, and retained activity under physiologically relevant salt, serum, and temperature conditions. In vivo, topical administration of AMP-hydrogel significantly reduced bacterial burden, accelerated wound closure, enhanced collagen deposition, and favorably modulated inflammatory cytokine responses compared with untreated controls.

Conclusions: The findings highlight bacteriophage holins as an underexplored source of antimicrobial peptides and demonstrate the potential of a novel AI-designed holin-derived peptide as a stable, biocompatible, and effective therapeutic candidate for managing MDR P. aeruginosa wound infections within a One Health framework addressing antimicrobial resistance.