Aims: Pholiota populnea is an edible macrofungus, distributed worldwide wherever cottonwood occurs. This study aimed to explore the potential antimicrobial properties of isolated lanostane triterpenes from P. populnea, namely pholiols A-D, E, G, H, J, L, and Q. Methods: A diverse range of microorganisms, including Gram-negative, and Gram-positive bacterial and fungal strains were employed to assess the antimicrobial activity of these natural triterpenes. The effects of the compounds on bacterial growth were determined by microdilution method. Results: Among the investigated compounds, pholiol C [3â-acetoxy-2á-(3-hydroxy-3-methylglutaroyloxy)-12â,25dihydroxylanosta-8-en-24-one] exhibited significant activity against Gram-positive bacteria, with minimum inhibitory concentrations (MIC) of 100 ěM against Streptococcus agalactiae, and 200 ěM against Staphylococcus aureus, the methicillin resistant S. aureus (MRSA), S. epidermidis, Enterococcus faecalis, and Bacillus subtilis. Additionally, pholiols G and E displayed notable inhibitory properties against the S. agalactiae strain, both exhibiting a MIC value of 200 ěM. Conclusion: These findings underscore the potential of the mushroom P. populnea as a noteworthy source of triterpenes possessing antimicrobial activity.  Kulcsszavak: pholiols, triterpenes, Pholiota populnea, antimicrobial activity