Investigating Sortase A inhibitory potential of herbal compounds using integrated computational and biochemical approaches

•The global health threat is growing because multi-drug-resistant bacteria are causing significant harm to mortality, morbidity, and healthcare costs.•ESKAPE pathogens are responsible for most nosocomial infections and can “escape” the biocidal action of antimicrobial agents.•The ESKAPE acronym stands for six pathogens namely Enterococcus faecium (E.faecalis), Staphylococcus aureus (S.aureus), Klebsiella pneumoniae (K.pneumoniae), Acinetobacter baumannii (A.baumanii), Pseudomonas aeruginosa (P.aeruginosa), and Enterobacter species (ESKAPE) which primarily increase the accountability of multi-drug resistance (MDR).•EfSrtAΔN59, a transpeptidase significant for the adhesion and virulence of Enterococcus faecalis (E. faecalis), presents an attractive target for disrupting biofilm formation–a key factor in persistent infections. Multi-drug resistance in bacteria is emerging as a major global health challenge, causing substantial harm in terms of mortality, morbidity, and financial strain on healthcare systems. These bacteria are constantly acquiring new virulence factors and drug-resistance mechanisms, which highlights the critical need for innovative antimicrobial medicines and identification of new therapeutic targets, such as Sortase A (EfSrtAΔN59). EfSrtAΔN59, a transpeptidase significant for the adhesion and virulence of Enterococcus faecalis (E. faecalis), presents an attractive target for disrupting biofilm formation—a key factor in persistent infections. This study investigates the inhibitory effects of two natural flavonoids- Rutin Trihydrate and Quercetin, on EfSrtAΔN59 and biofilm formation in E. faecalis. With in vitro enzymatic assays and biofilm quantification techniques, we demonstrate that both compounds significantly attenuate EfSrtAΔN59 activity, thereby hindering bacterial biofilm formation. Rutin Trihydrate and Quercetin exhibited strong binding affinities to the EfSrtAΔN59 enzyme, as confirmed by molecular docking and MD simulation studies. This was further substantiated by a notable reduction in biofilm biomass in bacterial cultures treated with these compounds. These findings highlight the potential of Rutin Trihydrate and Quercetin as promising candidates for the development of novel anti-virulence therapies aimed at mitigating E. faecalis infections, thereby offering a compelling alternative to traditional antibiotics.