This study explores the bioactivity potential of four indigenous Ganoderma species from northern Serbia (G. applanatum, G. lucidum, G. pfeifferi, and G. resinaceum), comparing antioxidant and anti-acetylcholinesterase (anti-AChE) properties. The mycochemical profile, including ethanol (EtOH), water (H2O), and chloroform (CHCl3) extracts, was analysed, revealing significant phenolic compounds, such as 2,5-dihydroxybenzoic acid. Highest total phenolic content (TP) was in EtOH extracts (205 - 380 mg GAE/g d.w.), and total carbohydrate content peaked in G. pfeifferi CHCl3 extracts (1895 ± 0.29 mg GluE/g d.w., 439 ± 0.42 mg FruE/g d.w.). DPPH and FRAP activities were prominent in G. applanatum H2O extracts, while CHCl3 extracts excelled in the NO assay, and EtOH extracts were superior in the ABTS assay. The most effective AChE inhibitors were polar extracts from G. pfeifferi. These findings emphasise the potential therapeutic use of Ganoderma species, particularly G. pfeifferi, in combating oxidative stress and acetylcholine deficiencies.