Trametes lactinea (T. lactinea), a macrofungus belonging to the family of Polyporaceae was investigated for its ability to inhibit hyaluronidase, lipoxygenase and xanthine oxidase activities in vitro. Results showed that extracts from the supernatant and mycelia of T. lactinea significantly inhibited the activities of hyaluronidase and lipoxygenase with more than 80% inhibition. The presence of total phenolics and flavonoids were also determined in the fungus extracts. The highest total phenolics and flavonoids were found in the supernatant of T. lactinea culture. Linear regression analysis carried out has found no correlation between the bioactive compounds (phenolics and flavonoids) and the inhibition of the enzyme activities, p > 0.05. The gas chromatography mass spectrometry (GCMS) analysis carried out has identified the presence of other bioactive compounds in the extracts that may act as the enzyme inhibitors.