The present study was aimed at to investigate the bioactive compounds from exudate of a fungus, Pleurotus ostreatus. Exudate as liquid droplets on the mycelium was checked for antimicrobial activity against bacterial strain, Bacillus subtilus. Biochemical techniques, thin layer chromatography, high performance liquid chromatography and gas chromatography/mass spectrometry, were employed to study fungal exudate. A total of fifteen different metabolites were detected in the sample by GC/MS. The detected metabolites could be classified into chemical groups of 2 esters, 4 alcohols, 3 benzoic acids, ketone, aldehyde, phenyl and amide groups containing compounds. Some of these may be classified into aroma containing compounds.