The present study mainly focused on identifying the efficiency of polypore mushrooms and exploring their medicinal value. In this experiment, the antioxidant and antimicrobial potentials of three mushrooms Trametes pubescens (Schumach.) Pilat, Spongipellis pachyodon (Pers.) Kotl. & Pouzar and Inonotus hispidus (Bull.) P. Karst. were tested along with their qualitative and quantitative analysis. The antioxidant and antimicrobial activities of methanolic crude extracts were determined using the Frap assay (ferric reducing antioxidant power) and agar well diffusion method respectively. The methanolic extracts of all three mushrooms revealed the presence of alkaloids, carbohydrates, proteins, phenolics, flavonoids, saponins, terpenoids, and cardiac glycosides. The antioxidant activity of Trametes pubescens, Spongipellis pachyodon, and Inonotus hispidus found to be 19.83±0.5, 38.5±0.8 and 39.5± 0.9 mg Fe (Ⅱ)/g of sample, respectively. The antimicrobial activity was tested against Staphylococcus aureus, Bacillus subtilis and Psuedomonas species. The inhibition zone was measured, in which all three species show maximum inhibition against Staphylococcus aureus. The correlation coefficient showed a positive relationship between total flavonoid content and antioxidant activity (r = 0.9). The Fourier Transformed infrared (FTIR) profiling indicates the presence of functional groups in these species for its pharmaceutical activities.