Antimicrobial Activity and Integrated In Silico Evaluation of Bioactive Constituents from Trametes versicolor against Clinically Isolated Pathogens

The increasing burden of antimicrobial resistance has intensified the search for structurally diverse antimicrobial agents from natural sources. This study evaluated the cultivation performance, extraction yield, phytochemical profile, antimicrobial activity, and computational properties of bioactive constituents associated with Trametes versicolor using an integrated experimental and in silico workflow. Fruiting bodies were cultivated on unsupplemented and supplemented sawdust substrates, extracted with methanol, ethanol, and water, and evaluated against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa , and Candida albicans by disc diffusion and broth dilution assays. Minimum inhibitory concentration, minimum bactericidal or fungicidal concentration, and dose response analyses were incorporated into the antimicrobial assessment. Qualitative phytochemical screening and gas chromatography mass spectrometry were used for chemical characterization. Candidate ligands were subsequently evaluated by physicochemical and medicinal chemistry screening, ADMET prediction, molecular docking against the ATPase domain of Staphylococcus aureus DNA gyrase B, density functional theory, and 200 ns molecular dynamics simulations. Supplemented sawdust increased fresh yield from 102.60 ± 3.58 g to 373.80 ± 5.21 g and biological efficiency from 11.10 ± 0.46% to 40.30 ± 0.56%. The aqueous extract gave the highest crude extraction yield, whereas methanol produced the highest overall inhibition across the antimicrobial panel. The largest inhibition zone was 22.33 ± 2.08 mm against S. aureus. The antimicrobial response depended significantly on extract, concentration, microorganism, and their interactions. GC MS profiling revealed a mixture dominated by terpenoid and oxygenated constituents. Among the computational ligands, CID 5281515 produced the most favourable docking score against GyrB at − 7.3 kcal/mol, followed by CID 91457 and CID 3084954 at − 7.1 kcal/mol. The four ligands subjected to DFT analysis showed HOMO LUMO gaps of 3.322 to 6.479 eV. Molecular dynamics trajectories distinguished the conformational behaviour of the complexes, with CID 5281515 showing the most consistent overall profile among the selected ligands. The integrated findings support further isolation, structural confirmation, and target based experimental evaluation of T. versicolor constituents as potential antimicrobial leads.