Introduction: Bioactive proteins and peptides generated from fruit, vegetables, meat or fish have great potential as functionalfood or substitutes for antibiotics. In recent years it has also been demonstrated that the fungus kingdom could be a source of thesecompounds. The study investigated the bioactivity of an extract of the lignicolous fungus Trametes versicolor and its hydrolysate.Material and Methods: The fungus was collected in a mixed forest in October, extracted and hydrolysed. To inspect the proteinand peptide profiles before and after hydrolysis, matrix-assisted laser desorption/ionisation–time-of-flight mass spectrometricanalysis was performed. To evaluate the antioxidant properties of the preparations, 2,2-diphenyl-1-picrylhydrazyl (DPPH●) and2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS●+) radical scavenging assays were used. The activity of the fungusextract and hydrolysate against Aeromonas veronii, Bacillus cereus, Enterococcus faecalis, Enterococcus faecium, Escherichiacoli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella Typhimurium, Staphylococcus aureus, Staphylococcusepidermidis, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis was determined by the minimuminhibitory concentration and minimum bactericidal concentration values. Results: The extract and its hydrolysate showed almost100% ABTS●+ and DPPH● radical scavenging with a low half maximal inhibitory concentration. The water extract and hydrolysateof T. versicolor exhibited antimicrobial activity against two S. aureus strains, E. coli, P. aeruginosa and Salmonella Typhimurium.Conclusion: These results provide compelling evidence that the analysed fungus extract and its hydrolysate hold promise withtheir antibacterial and antioxidant properties.