Bioactive Potential of Balkan <i>Fomes fomentarius</i> Strains: Novel Insights into Comparative Mycochemical Composition, Antioxidant, Anti-Acetylcholinesterase, and Antiproliferative Activities
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Fomes fomentarius (L.) Fr., known for its medicinal properties since the 15th century, has been traditionally used across Europe, including the Balkan region, for tinder, spiritual practices, wound healing, and hemostasis. This study investigates the mycochemical profile of three Balkan strains, focusing on micro- and macroelements, polyamines (PAs), and phenolic compounds, alongside the bioactivity of hot water (H2O), chloroform (CHCl3), hydroethanolic (EtOH), and hydromethanolic (MeOH) extracts. Micro- and macroelements were analyzed via atomic absorption spectrophotometry (AAS), while PAs were quantified using HPLC-FD. Phenolic profiles were determined by LC-MS/MS, with total phenolic content (TPC) assessed using the Folin-Ciocalteu method. Antioxidant potential was evaluated through DPPH, ABTS, NO scavenging, lipid peroxidation inhibition, and FRAP assays, in addition to anti-acetylcholinesterase and antiproliferative activity assessments. This study represents the first investigation of polyamine profiles in F. fomentarius, with total PA levels ranging from 32.67 to 2910.09 nmol/g d.w. The Bosnian strain exhibited the highest PA levels, with spermidine (SPD) concentrations between 899.96 to 2910.09 nmol/g d.w. LC-MS/MS analysis identified several bioactive phenolics, including amentoflavone, baicalein, chrysoeriol, esculetin, and scopoletin, which are reported here for the first time in this fungus. H2O and EtOH extracts from Croatia and Serbia showed higher TPC content, correlating with notable antioxidant activity. EtOH and MeOH extracts demonstrated significant anti-acetylcholinesterase and antiproliferative activities, emphasizing their medicinal potential. Our findings highlight the therapeutic potential of polar extracts from Balkan F. fomentarius as natural antioxidants with applications in oxidative stress-related chronic diseases.