Nisin, a promising antimicrobial peptide, forestalls the methicillin-resistant Staphylococcus aureus biofilm network via reactive oxygen species generation.

Staphylococcal infections have been reported to be a significant global threat to the effective management of public healthcare due to their drug resistance property. This attribute has further been complicated by their robust biofilm-forming potential. This escalating threat of biofilm-associated infections necessitates innovative and promising therapeutic strategies. Hence, in the present study, the biofilm threat of methicillin-resistant Staphylococcus aureus (MRSA) has been challenged by Nisin, a natural lantibiotic produced by Lactococcus lactis. This compound showed a promising antibacterial effect with minimum inhibitory concentrations (MICs) of 150 µg/ml against MRSA. Furthermore, a series of experiments has been conducted to confirm the antibiofilm potential of Nisin against MRSA. Towards this direction, the sub-MIC dose of Nisin (40 µg/mL) was found to inhibit biofilm formation by ~ 51% for MRSA. To support this finding, extracellular polymeric substance (EPS) was measured under the Nisin-treated and untreated conditions of MRSA. It was observed that Nisin could destabilise the MRSA biofilm by reducing the EPS production to an extent of ~ 55%. Mechanistic studies further demonstrated that Nisin was found to increase the intracellular accumulation of reactive oxygen species (ROS), which could lead to the alteration of cell membrane permeability. Additionally, Nisin attenuated staphyloxanthin production (~ 54%), hemolytic ability (~ 26%), and fibrinogen clumping ability (~ 27%) of MRSA, suggesting its interference in the virulence profile of MRSA. Collectively, these findings suggest Nisin's dual role as a promising Staphylococcal biofilm inhibitor and virulence factor suppressor, making it a viable option for the treatment of MRSA-linked infections. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-025-04597-8.