Engineering Dual-Target Chimeric Lysins for Synergistic Eradication of Porphyromonas gingivalis

Periodontitis is a chronic inflammatory disease driven by complex subgingival multispecies biofilms, in which Porphyromonas gingivalis plays a central role in coordinating microbial interactions. Together with host-associated factors, these microbial communities create dual constraints that limit antimicrobial efficacy. In this study, we engineered a library of recombinant lysins by fusing membrane-destabilizing peptides to the periodontal pathogen-derived lysin LysPd078 and identified four optimized variants (PlyPd06, PlyPd19, PlyPd27, and PlyPd44) with enhanced salt tolerance, environmental stability, and bactericidal activity. In a clinically derived polymicrobial oral biofilm, PlyPd44 at only 25 g/mL eradicated 96.6% of P. gingivalis. In a humanized oral microbiota mouse model of periodontitis, selected PlyPds significantly reduced inflammation and inhibited alveolar bone loss compared with LysPd078 and minocycline. Collectively, these results establish membrane-destabilizing peptide-enabled lysins as a promising platform for developing microenvironment-adapted precision antimicrobials for periodontitis.