Mushrooms have been essential to the human diet because they contain balanced chemical components and some biologically active substances. In this work, we investigated the phenolics, essential oils, metal contents, antioxidant, antibacterial, DNA protective, and enzyme inhibition activities for Clitocybe geotropa, Ramaria aurea, Rhizopogon luteolus (RL), Russula delica (RD), Verpa bohemica, and Marasmius oreades mushrooms. Results exhibited a higher content for citric and succinic acids in all tested kinds. Further, we detected a high content of cis‐9‐oleic acid, linoleate, and cis‐11‐eicosanoate. All mushroom species contain a significant percentage of both Cu and Zn. Moreover, RL and RD recorded the highest phenolic and flavonoid contents. Furthermore, all samples showed standard to good antioxidant activity, and the same is true for the antibacterial and DNA protective activities. Enzyme inhibition activity was generally high and significantly higher against the urease than the thiourea. We applied molecular docking between the highest phenolic molecules with the urease to determine the mushroom extracts’ high inhibition mechanism. In conclusion, all mushroom species revealed a variety in chemical content that is probably related to their multi‐bioactivity.