This study engineered a recombinant lysostaphin (Lst) variant, Lst-ELP(V)₉₀, as a therapeutic candidate for prosthetic explant biofilm infections caused by methicillin-resistant Staphylococcus aureus (MRSA). Compared with the wild-type (WT) lysostaphin, the recombinant construct exhibited markedly improved stability and attenuated immunogenicity, establishing a robust foundation for downstream drug development.