Evaluation of the phenolic profile and biological activities of Boletus speciosus extracts optimized with response surface methodology and artificial neural networks-genetic algorithm

In this study, the antioxidant, anticholinesterase, and antiproliferative activities, as well as the phenolic component profiles, of extracts obtained under optimum conditions from Boletus speciosus using Response Surface Methodology (RSM) and Artificial Neural Networks-Genetic Algorithm (ANN-GA) approaches were comprehensively evaluated. Antioxidant capacity was determined by total antioxidant level (TAS), total oxidant level (TOS), oxidative stress index (OSI), DPPH radical scavenging activity, and FRAP reducing power tests. Anticholinesterase activity was evaluated through acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition, while the antiproliferative effect was investigated in vitro in A549, MCF-7, and DU-145 cell lines at concentrations ranging from 25 to 200 µg/mL. Quantitative analysis of phenolic components was performed using LC-MS/MS. The results showed that extracts obtained by the ANN-GA method exhibited significantly higher TAS, DPPH, and FRAP values, and lower TOS and OSI values ​​compared to RSM extracts. In anticholinesterase analyses, ANN-GA extracts were found to have a stronger inhibitory effect on both AChE and BChE. In antiproliferative analyses, it was determined that ANN-GA extracts caused a more pronounced dose-dependent decrease in cell viability in all cell lines. LC-MS/MS analyses revealed that various phenolic compounds, especially gallic acid, protocatechuic acid, caffeic acid, quercetin, and catechin hydrate, were present in higher amounts in ANN-GA extracts. In conclusion, it was determined that B. speciosus has a rich phenolic content, and that the ANN-GA optimization approach, in particular, enhanced the antioxidant, anticholinesterase, and antiproliferative activities by increasing the extraction of phenolic compounds. These findings demonstrate that B. speciosus is a functionally and biologically valuable natural resource and offers significant potential for advanced biomedical research.