The global rise of multidrug-resistant Klebsiella pneumoniae underscores the urgent need for alternative therapeutic strategies. Bacteriophage-derived depolymerases have emerged as promising antimicrobial factors, selectively degrading bacterial capsules and impairing key pathogenic traits. We characterize a novel depolymerase, PRA33gp45, associated with the structural protein of bacteriophage vBKpnPPRA33. Bioinformatic structural analyses predicted endo-N-acetyl neuraminidase-like activity and canonical depolymerase domain architecture. The recombinant PRA33gp45 specifically hydrolysed capsular polysaccharides (CPS) of K27-serotype K. pneumoniae and produced characteristic halo zones on bacterial lawns, confirming its enzymatic activity. Capsule staining demonstrated rapid and progressive capsule degradation within 120 minutes of treatment. PRA33gp45 significantly inhibited biofilm formation, disrupted mature biofilms, and altered biofilm architecture as visualized by confocal microscopy. Depolymerase pre-treatment markedly reduced K. pneumoniae survival within A549 human lung epithelial cells, without exhibiting any cytotoxic effect and sensitized bacteria to complement-mediated killing in human serum. Finally, PRA33gp45 treatment of K. pneumoniae lowers morbidity and mortality in the Galleria mellonella larvae model. Collectively, these findings identify PRA33gp45 as a novel and highly specific depolymerase that diminishes K. pneumoniae virulence by targeting its protective capsule, impairing persistence as biofilm, and enhancing innate immune clearance. Its safety and efficacy suggest potential as an antimicrobial or adjuvant therapeutic agent against K27-type K. pneumoniae infections, particularly in the context of multidrug resistance and emerging pathogens.
Pelka et al. (Fri,) studied this question.