Mushrooms of the genera Hericium and Inonotus are valuable sources of bioactive polyphenols with notable antioxidant and antimicrobial properties. This study examined the total polyphenol content , antioxidant potential, and antimicrobial activity of ethanol and methanol extracts obtained from submerged mycelial cultures of Hericium erinaceus 2530 and Inonotus obliquus 2512. The total polyphenol content (TPC) was quantified using the Folin–Ciocalteu method, and antioxidant capacity was assessed by the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay. Fourier Transform Infrared spectroscopy was applied to identify the principal functional groups of phenolic constituents. Marked strain-dependent variations were observed in both polyphenol content and antioxidant performance, with Inonotus obliquus 2512 showing the highest activity and Hericium erinaceus 2530 demonstrating moderate but significant effects. Both species exhibited measurable antibacterial activity against Staphylococcus aureus and Escherichia coli. These findings underscore the potential of submerged mushroom cultures as sustainable sources of natural antioxidants and antimicrobial agents for prospective nutraceutical and pharmaceutical applications.