Natural compounds with biological activity are normally present in plants, mushrooms and their natural sources. Applied mycology is one of the most stimulating and rapidly evolving areas of the biological sciences. Hence the present study focussed on exploring Microporus affinis (Blume & T. Nees) Kunt., the least explored and edible bracket fungus. Chemical characterization by GC-MS analysis resulted in the presence of 47 bioactive compounds. 9, 12- Octadecadienoic acid (Z,Z)- methyl ester, Ergosterol, Monolinolein, Thiacremonone, Stellasterol, n- Hexadecanoic acid, Ribitol, Maltol etc., were the leading compounds. Because of the presence of various bioactive compounds which have been already reported to possess antitumor, antioxidant and anticancer activities, M. affinis extract has been tested for in vitro anticancer efficacy on DLD1 cell lines (cultured in DMEM medium) using MTT assay. It resulted in the decrease of percentage of viability as the increase in concentration of the extract. Apoptosis was determined by using Acridine orange and Ethidium bromide staining. Thus, the taxa, M. affinis can be recommended for further anticancer assays for validation.