Wild mushroom Tylopilus felleus antioxidant, anticholinesterase and oxidant status and heavy metal contents

Abstract

Mushrooms play essential roles in natural ecosystems due to their ecological functions and bioactive potential, and many species are also valued for nutritional and medicinal purposes. In the present study, the antioxidant and oxidant status, anticholinesterase activity, and heavy metal contents of the fruiting bodies of the wild mushroom Tylopilus felleus (Bull.) P. Karst. were investigated. Total antioxidant status (TAS) and total oxidant status (TOS) were determined using Rel Assay commercial kits, and the oxidative stress index (OSI) was calculated accordingly. Anticholinesterase activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were evaluated based on the Ellman method, with galantamine used as a reference inhibitor. Heavy metal contents, including Fe, Zn, Cu, Pb, and Ni, were quantified using an atomic absorption spectrophotometer (Agilent 240FS AA). The TAS, TOS, and OSI values of T. felleus were found to be 3.427±0.147, 20.512±0.266, and 0.601±0.031, respectively. Elemental analysis revealed Fe, Zn, Cu, Pb, and Ni contents of 119.57±12.78, 21.07±1.69, 44.10±2.07, 2.89±0.47, and 0.82±0.011 mg/kg, respectively, all of which were within ranges reported in the literature. The mushroom extract exhibited moderate inhibitory activity against AChE (IC₅₀: 192.16 ± 3.15 μg/mL) and BChE (IC₅₀: 236.99±1.22 μg/mL) compared to galantamine. Overall, the findings suggest that T. felleus represents a potential natural source of antioxidant compounds with supplementary anticholinesterase activity.

Keywords

• Antioxidant
• Anticholinesterase
• Bitter bolete
• Medicinal mushroom
• Wild mushroom

References

1. Ahmed, A., Taj, M.K., Yongliang, Z., Taj, I., Hassani, T.M., Sajid, M., Ahmed, Z., Azizullah, A., Mengal, M.A., & Liping, S. (2015). Chemical composition and antioxidant properties of eight wild edible Boletaceae mushrooms growing in Yunnan Province of China. Jokull Journal, 65(12), 146–167.
2. Andrade, J.M., Pachar, P., Trujillo, L., & Cartuche, L. (2022). Suillin: A mixed type acetylcholinesterase inhibitor from Suillus luteus which is used by Saraguros indigenous, southern Ecuador. PLoS One, 17(5), e0268292. https://doi.org/10.1371/journal.pone.0268292
3. Baba, H., Sevindik, M., Gürgen, A., Krupodorova, T., & Akata, I. (2025). Pharmacological potential of Russula grata: Antioxidant, neuroprotective, and anticancer properties. Studies in Fungi, 10(1), e027. https://doi.org/10.48130/sif-0025-0027
4. Bal, C. (2018). A study on antioxidant properties of Gyrodon lividus. Eurasian Journal of Forest Science, 6(2), 40–43. https://doi.org/10.31195/ejejfs.425071
5. Bal, C., Sevindik, M., Akgul, H., & Selamoglu, Z. (2019). Oxidative stress index and antioxidant capacity of Lepista nuda collected from Gaziantep/Turkey. Sigma, 37(1), 1–5.
6. Dundar, A., Okumus, V., Ozdemir, S., Celik, K. S., Boğa, M., & Ozcagli, E. (2016). Determination of cytotoxic, anticholinesterase, antioxidant and antimicrobial activities of some wild mushroom species. Cogent Food & Agriculture, 2(1), 1178060. https://doi.org/10.1080/23311932.2016.1178060
7. Ellman, G.L., Courtney, K.D., Andres, V. Jr., & Featherstone, R.M. (1961). A new and rapid colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology, 7(2), 88–95. https://doi.org/10.1016/0006-2952(61)90145-9
8. Emsen, B., Guven, B., & Kaya, A. (2019). Antioxidant and antigenotoxic potential of Lycoperdon molle Pers., a wild edible mushroom. Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 22(5), 724–732. https://doi.org/10.18016/ksutarimdoga.vi.523756

9. Erel, O. (2005). A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry, 38(12), 1103–1111. https://doi.org/10.1016/j.clinbiochem.2005.08.008
10. Erel, O. (2004). A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry, 37(4), 277–285. https://doi.org/10.1016/j.clinbiochem.2003.11.015
11. Falandysz, J., Kunito, T., Kubota, R., Bielawski, L., Mazur, A., Falandysz, J.J., et al. (2007). Selected elements in brown birch scaber stalk Leccinum scabrum. Journal of Environmental Science and Health A, 42, 2081–2088. https://doi.org/10.1080/10934520701626993
12. Falandysz, J., Kunito, T., Kubota, R., Gucia, M., Mazur, A., Falandysz, J.J., et al. (2008). Some mineral constituents of parasol mushroom (Macrolepiota procera). Journal of Environmental Science and Health B, 43, 187–192. https://doi.org/10.1080/03601230701795247
13. Gürgen, A., & Sevindik, M. (2022). Application of artificial neural network coupling multiobjective particle swarm optimization algorithm to optimize Pleurotus ostreatus extraction parameters. Journal of Food Processing and Preservation, 46(11), e16949. https://doi.org/10.1111/jfpp.16949
14. Gürgen, A., & Sevindik, M. (2025). Optimization of biological activities of Agaricus species: An artificial intelligence assisted approach. Scientific Reports, 15(1), 23147. https://doi.org/10.1038/s41598-025-08118-8
15. Hamed, A.A., El-Shiekh, R.A., Mohamed, O.G., Aboutabl, E.A., Fathy, F.I., Fawzy, G.A., et al. (2023). Cholinesterase inhibitors from an endophytic fungus Aspergillus niveus Fv-er401: Metabolomics, isolation and molecular docking. Molecules, 28(6), 2559. https://doi.org/10.3390/molecules28062559
16. Karaltı, İ., Eraslan, E.C., Sarıdoğan, B.G. Ö., Akata, I., & Sevindik, M. (2022). Total antioxidant, antimicrobial, antiproliferative potentials and element contents of wild mushroom Candolleomyces candolleanus (Agaricomycetes) from Turkey. International Journal of Medicinal Mushrooms, 24(12), 69–76. https://doi.org/10.1615/intjmedmushrooms.2022045389
17. Korkmaz, A.F., Gürgen, A., Krupodorova, T., Sevindik, M., & Akata, I. (2025). Artificial intelligence-assisted optimization of extraction enhances the biological activity of Phylloporia ribis. Scientific Reports, 15(1), 41206. https://doi.org/10.1038/s41598-025-25130-0
18. Krupodorova, T., & Sevindik, M. (2020). Antioxidant potential and some mineral contents of wild edible mushroom Ramaria stricta. AgroLife Scientific Journal, 9, 186–191.
19. Lee, E.J., & Jang, H.D. (2004). Antioxidant activity and protective effect of five edible mushrooms on oxidative DNA damage. Food Science and Biotechnology, 13(4), 443-449
20. Lee, J.E., Lee, I.S., Kim, K.C., Yoo, I.D., & Yang, H.M. (2017). ROS scavenging and anti-wrinkle effects of clitocybin A isolated from the mycelium of the mushroom Clitocybe aurantiaca. Journal of Microbiology and Biotechnology, 27(5), 933–938. https://doi.org/10.4014/jmb.1702.02050
21. Ma, L., Chen, H., Dong, P., & Lu, X. (2013). Anti-inflammatory and anticancer activities of extracts and compounds from the mushroom Inonotus obliquus. Food Chemistry, 139(1–4), 503–508. https://doi.org/10.1016/j.foodchem.2013.01.030
22. Macáková, K., Opletal, L., Polášek, M., Samková, V., & Jahodář, L. (2009). Free-radical scavenging activity of some European Boletales. Natural Product Communications, 4(2), 1934578X0900400219.
23. Manzi, P., Aguzzi, A., & Pizzoferrato, L. (2001). Nutritional value of mushrooms widely consumed in Italy. Food Chemistry, 73(3), 321–325. https://doi.org/10.1016/S0308-8146(00)00304-6
24. Mišković, J., Rašeta, M., Čapelja, E., Krsmanović, N., Novaković, A., Janjušević, L., & Karaman, M. (2021). Mushroom species Stereum hirsutum as a natural source of phenolics and fatty acids as antioxidants and acetylcholinesterase inhibitors. Chemistry & Biodiversity, 18(11), e2100409. https://doi.org/10.1002/cbdv.202100409
25. Mleczek, M., Budka, A., Siwulski, M., Mleczek, P., Budzyńska, S., Proch, J., et al. (2021). A comparison of toxic and essential elements in edible wild and cultivated mushroom species. European Food Research and Technology, 247(5), 1249–1262. https://doi.org/10.1007/s00217-021-03706-0
26. Murray, A.P., Biscussi, B., Cavallaro, V., Donozo, M., & Rodriguez, S.A. (2024). Naturally occurring cholinesterase inhibitors from plants, fungi, algae, and animals: A review of the most effective inhibitors reported in 2012 2022. Current Neuropharmacology, 22(10), 1621 1649. https://doi.org/10.2174/1570159X21666230623105929
27. Nguyen, T.K., Im, K.H., Choi, J., Shin, P.G., & Lee, T.S. (2016). Evaluation of antioxidant, anticholinesterase, and anti-inflammatory effects of culinary mushroom Pleurotus pulmonarius. Mycobiology, 44(4), 291–301. https://doi.org/10.5941/MYCO.2016.44.4.291
28. Nikkarinen, M., & Mertanen, E. (2004). Impact of geological origin on trace element composition of edible mushrooms. Journal of Food Composition and Analysis, 17(3 4), 301 310. https://doi.org/10.1016/j.jfca.2004.03.013
29. Nitha, B., Meera, C.R., & Janardhanan, K.K. (2007). Anti-inflammatory and antitumour activities of cultured mycelium of morel mushroom, Morchella esculenta. Current Science, 92(2), 235–239.
30. Pedneault, K., Angers, P., Gosselin, A., & Tweddell, R.J. (2008). Fatty acid profiles of polar and neutral lipids of ten species of higher basidiomycetes indigenous to eastern Canada. Mycological Research, 112(12), 1428–1434. https://doi.org/10.1016/j.mycres.2008.06.026
31. Ren, L., Perera, C., & Hemar, Y. (2012). Antitumor activity of mushroom polysaccharides: A review. Food & Function, 3(11), 1118–1130. https://doi.org/10.1039/c2fo10279j
32. Sarikurkcu, C., Copur, M., Yildiz, D., & Akata, I. (2011). Metal concentration of wild edible mushrooms in Soguksu National Park in Turkey. Food Chemistry, 128(3), 731 734. https://doi.org/10.1016/j.foodchem.2011.03.097
33. Sevindik, M. (2020). Antioxidant and antimicrobial capacity of Lactifluus rugatus and its antiproliferative activity on A549 cells. Indian Journal of Traditional Knowledge, 19(2), 423–427.
34. Sevindik, M. (2021). Anticancer, antimicrobial, antioxidant and DNA protective potential of mushroom Leucopaxillus gentianeus (Quél.) Kotl. Indian Journal of Experimental Biology, 59(5), 310–315.
35. Sevindik, M. (2025). Evaluation of antioxidant, anticholinesterase and antiproliferative potential of Artemisia herba-alba by artificial intelligence-assisted extraction optimization. Scientific Reports, 15(1), 37985. https://doi.org/10.1038/s41598-025-21832-7
36. Sevindik, M., & Akata, I. (2019). Antioxidant and oxidant potentials and element content of edible wild mushroom Helvella leucopus. Indian Journal of Natural Products and Resources, 10(4), 266–271.
37. Sevindik, M., Akgul, H., Selamoglu, Z., & Braidy, N. (2020). Antioxidant and antigenotoxic potential of Infundibulicybe geotropa mushroom collected from northwestern Turkey. Oxidative Medicine and Cellular Longevity, 2020, 5620484. https://doi.org/10.1155/2020/5620484
38. Sevindik, M., Bal, C., Eraslan, E.C., Uysal, I., & Mohammed, F.S. (2023). Medicinal mushrooms: A comprehensive study on their antiviral potential. Prospects in Pharmaceutical Sciences, 21(2), 42–56. https://doi.org/10.56782/pps.141
39. Sevindik, M., Eraslan, E.C., Krupodorova, T.A., Krakhmalnyi, M., Uysal, I., Bal, C., & Wasser, S.P. (2025). The role of medicinal mushrooms in cancer treatment: Bioactive compounds and therapeutic potential. International Journal of Medicinal Mushrooms, 27(12), 1 24. https://doi.org/10.1615/intjmedmushrooms.2025060581
40. Sevindik, M., Gürgen, A., Khassanov, V.T., & Bal, C. (2024). Biological activities of ethanol extracts of Hericium erinaceus obtained as a result of optimization analysis. Foods, 13(10), 1560. https://doi.org/10.3390/foods13101560
41. Siwulski, M., Rzymski, P., Budka, A., Kalač, P., Budzyńska, S., Dawidowicz, L., et al. (2019). The effect of different substrates on the growth of six cultivated mushroom species and composition of macro and trace elements in their fruiting bodies. European Food Research and Technology, 245(2), 419–431. https://doi.org/10.1007/s00217-018-3174-5
42. Sutthibutpong, T., Posansee, K., Liangruksa, M., Termsaithong, T., Piyayotai, S., Phitsuwan, P., et al. (2024). Combining deep learning and structural modeling to identify potential acetylcholinesterase inhibitors from Hericium erinaceus. ACS Omega, 9(14), 16311 16321. https://doi.org/10.1021/acsomega.3c10459
43. Tel, G., Ozturk, M., Duru, M.E., & Turkoglu, A. (2015). Antioxidant and anticholinesterase activities of five wild mushroom species with total bioactive contents. Pharmaceutical Biology, 53(6), 824–830. https://doi.org/10.3109/13880209.2014.943245
44. Ünal, O., Gürgen, A., Krupodorova, T., Sevindik, M., Kabaktepe, Ş., & Akata, I. (2025). Optimization of Phellinus hartigii extracts: Biological activities and phenolic content analysis. BMC Complementary Medicine and Therapies, 25(1), 113. https://doi.org/10.1186/s12906-025-04851-9
45. Valverde, M.E., Hernández-Pérez, T., & Paredes-López, O. (2015). Edible mushrooms: Improving human health and promoting quality life. International Journal of Microbiology, 2015, 376387. https://doi.org/10.1155/2015/376387
46. Vetter, J. (1990). Mineral element content of edible and poisonous macrofungi. Acta Alimentaria, 19(1), 27–40.
47. Wang, Y., Mo, Y., Li, D., Xiang, C., Jiang, Z., & Wang, J. (2019). The main factors inducing postharvest lignification in king oyster mushrooms (Pleurotus eryngii): Wounding and ROS-mediated senescence. Food Chemistry, 301, 125224. https://doi.org/10.1016/j.foodchem.2019.125224
48. Wei, Y.M., Yang, L., Wang, H., Cai, C.H., Chen, Z.B., Chen, H.Q., et al. (2022). Triterpenoids as bivalent and dual inhibitors of acetylcholinesterase/butyrylcholinesterase from the fruiting bodies of Inonotus obliquus. Phytochemistry, 200, 113182. https://doi.org/10.1016/j.phytochem.2022.113182
49. Wu, D.M., Duan, W.Q., Liu, Y., & Cen, Y. (2010). Anti-inflammatory effect of the polysaccharides of golden needle mushroom in burned rats. International Journal of Biological Macromolecules, 46(1), 100–103. https://doi.org/10.1016/j.ijbiomac.2009.10.013
50. Wu, W.T., Hsu, T.H., Lee, C.H., & Lo, H.C. (2020). Fruiting bodies of Chinese caterpillar mushroom Ophiocordyceps sinensis (Ascomycetes) alleviate diabetes-associated oxidative stress. International Journal of Medicinal Mushrooms, 22(1), 15 29. https://doi.org/10.1615/intjmedmushrooms.2019033275