Library Subscription: Guest
Begell Digital Portal Begell Digital Library eBooks Journals References & Proceedings Research Collections
International Journal of Medicinal Mushrooms
IF: 1.423 5-Year IF: 1.525 SJR: 0.431 SNIP: 0.661 CiteScore™: 1.38

ISSN Print: 1521-9437
ISSN Online: 1940-4344

International Journal of Medicinal Mushrooms

DOI: 10.1615/IntJMedMushrooms.2019031508
pages 755-763

Bioactivity-Guided Isolation of Acetylcholinesterase Inhibitor from Ganoderma mediosinense (Agaricomycetes)

Arshdeep Kaur
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala, Punjab, India
Kudrat Randhawa
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala, Punjab, India
Varinder Singh
Chitkara College of Pharmacy, Chitkara University, Punjab, India
Richa Shri
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala, Punjab, India


The involvement of high acetylcholinesterase (AChE) activity and oxidative stress in the brain is well documented in the progression of dementia. Thus, finding a drug with both AChE-inhibitory and antioxidant activities could be beneficial in treating dementia. We previously reported the AChE-inhibitory and antioxidant-mediated antiamnestic effects of a hydromethanol extract from Ganoderma mediosinense (HME), which we evaluated using in vitro and in vivo models. Mycochemical screening of HME showed the presence of phenols. Building on those findings, this study was designed to isolate the compound responsible for the AChE-inhibitory and antioxidant activities of G. mediosinense. The HME was fractionated sequentially with solvents−namely, hexane, chloroform, and ethyl acetate. The prepared fractions were evaluated by using Ellman and DPPH-inhibitory assays in vitro. Among the fractions, the ethyl acetate fraction showed appreciable AChE-inhibitory (half-maximal inhibitory concentration [IC50] 0.81 ± 0.04 mg/mL) and DPPH scavenging (IC50 2.04 ± 0.77 μg/mL) activities; therefore it was subjected to flash chromatography, which separated 9 subfractions. Subfraction 7 showed marked activity in inhibiting AChE (IC50 0.10 ± 0.02 mg/mL) and free radicals (IC50 1.22 ± 0.04 μg/mL). Purification of subfraction 7 yielded a white compound, AK1. This was characterized as gallic acid by using Fourier-transform infrared, nuclear magnetic resonance, and mass spectral studies. This study shows that gallic acid, a well-recognized phenolic acid, is responsible for the AChE-inhibitory and DPPH scavenging activities of G. mediosinense.


  1. Awasthi M, Singh S, Pandey VP, Dwivedi UN. Alzheimer's disease: an overview of amyloid beta dependent pathogenesis and its therapeutic implications along with in silico approaches emphasizing the role of natural products. J Neurol Sci. 2016;361:256-71.

  2. Auld DS, Kornecook TJ, Bastianetto S, Quirion R. Alzheimer's disease and the basal forebrain cholinergic system: relations to P-amyloid peptides, cognition, and treatment strategies. Prog Neurobiol. 2002;68(3):209-45.

  3. Colovic MB, Krstic DZ, Lazarevic-Pasti TD, Bondzic AM, Vasic VM. Acetylcholinesterase inhibitors: pharmacology and toxicology. Curr Neuropharmacol. 2013;11(3):315-35.

  4. Singh M, Kaur M, Kukreja H, Chugh R, Silakari O, Singh D. Acetylcholinesterase inhibitors as Alzheimer therapy: from nerve toxins to neuroprotection. Eur J Med Chem. 2013;70:165-88.

  5. Wachtel-Galor S, Yuen J, Buswell JA, Benzie IFF. Ganoderma lucidum (lingzhi or reishi): a medicinal mushroom. In: Benzie IFF, Wachtel-Galor S, editors. Herbal medicine: biomolecular and clinical aspects. 2nd ed. Boca Raton (FL): CRC Press; 2011.

  6. Chang ST, Buswell JA. Mushroom nutraceuticals. World J Microbiol Biotechnol. 1996;12(5):473-6.

  7. Kabir Y, Kimura S, Tamura T. Dietary effect of Ganoderma lucidum mushroom on blood pressure and lipid levels in spontaneously hypertensive rats (SHR). J Nutr Sci Vitaminol. 1988;34(4):433-8.

  8. Soccol CR, Bissoqui LY, Rodrigues C, Rubel R, Sella SR, Leifa F, de Souza Vandenberghe LP, Soccol VT. Pharmacological properties of biocompounds from spores of the lingzhi or reishi medicinal mushroom Ganoderma lucidum (Agaricomycetes): a review. Int J Med Mushrooms. 2016;18(9):757-67.

  9. Gerber AL, Smania A Jr, Monache FD, Biacchi N Jr, Smania ED. Triterpenes and sterols from Ganoderma australe (Fr.) Pat. (Aphyllophoromycetidae). Int J Med Mushrooms. 2000;2(4):303-11.

  10. Shi Y, Sun J, He H, Guo H, Zhang S. Hepatoprotective effects of Ganoderma lucidum peptides against D-galactosamine-induced liver injury in mice. J Ethnopharmacol. 2008;117(3):415-9.

  11. Tie L, Yang HQ, An Y, Liu SQ, Han J, Xu Y, Hu M, Li WD, Chen AF, Lin ZB, Li XJ. Ganoderma lucidum polysaccharide accelerates refractory wound healing by inhibition of mitochondrial oxidative stress in type 1 diabetes. Cell Physiol Biochem. 2012;29(3-4):583-94.

  12. Kaur R, Singh V, Shri R. Anti-amnesic effects of Ganoderma species: a possible cholinergic and antioxidant mechanism. Biomed Pharmacother. 2017;92:1055-61.

  13. Farnsworth NR. Biological and phytochemical screening of plants. J Pharm Sci. 1966;55(3):225-76.

  14. Ellman GL, Courtney KD, Andres V Jr, Feather-stone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7(2):88-95.

  15. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181:1199-200.

  16. Yiannopoulou KG, Papageorgiou SG. Current and future treatments for Alzheimer's disease. Ther Adv Neurol Disord. 2013;6(1):19-33.

  17. Francis PT, Palmer AM, Snape M, Wilcock GK. The cholinergic hypothesis of Alzheimer's disease: a review of progress. J Neurol Neurosurg Psychiatry. 1999;66(2):137-47.

  18. Pinto T, Lanctot KL, Herrmann N. Revisiting the cholinergic hypothesis of behavioral and psychological symptoms in dementia of the Alzheimer's type. Ageing Res Rev. 2011;10(4):404-12.

  19. Bennett S, Grant MM, Aldred S. Oxidative stress in vascular dementia and Alzheimer's disease: a common pathology. J Alzheimers Dis. 2009;17(2):245-57.

  20. Gella A, Durany N. Oxidative stress in Alzheimer disease. Cell Adh Migr. 2009;3(1):88-93.

  21. Garcia-Blanco A, Baquero M, Vento M, Gil E, Bataller L, Chafer-Pericas C. Potential oxidative stress biomarkers of mild cognitive impairment due to Alzheimer disease. J Neurol Sci. 2017;373:295-302.

  22. Luca M, Luca A, Calandra C. The role of oxidative damage in the pathogenesis and progression of Alzheimer's disease and vascular dementia. Oxid Med Cell Longev. 2015;2015:504678.

  23. Huang WJ, Zhang XI, Chen WW. Role of oxidative stress in Alzheimer's disease. Biomed Rep. 2016;4(5):519-22.

  24. Chanwitheesuk A, Teerawutgulrag A, Kilburn JD, Rakariyatham N. Antimicrobial gallic acid from Caesalpinia mimosoides Lamk. Food Chem. 2007;100(3):1044-8.

  25. Sarria-Villa RA, Gallo-Corredor JA, Paez MI. Isolation of catechin and gallic acid from Colombian bark of Pinus patula. Chem Sci J. 2017;8:174.

  26. Veljovic S, Veljovic M, Nikicevic N, Despotovic S, Radulovic S, Niksic M, Filipovic L. Chemical composition, anti-proliferative and antioxidant activity of differently processed Ganoderma lucidum ethanol extracts. J Food Sci Technol. 2017;54(5):1312-20.

  27. Kim MY, Seguin P, Ahn JK, Kim JJ, Chun SC, Kim EH, Seo SH, Kang EY, Kim SL, Park YJ, Ro HM. Phenolic compound concentration and antioxidant activities of edible and medicinal mushrooms from Korea. J Agric Food Chem. 2008;56:7265-70.

  28. Lin S, Ching LT, Ke X, Cheung PC. Comparison of the composition and antioxidant activities of phenolics from the fruiting bodies of cultivated Asian culinary-medicinal mushrooms. Int J Med Mushrooms. 2016;18(10):871-81.

  29. Gaur T, Rao PB. Analysis of antibacterial activity and bioactive compounds of the giant mushroom, Macrocybe gigantea (Agaricomycetes), from India. Int J Med Mushrooms. 2017;19(12):1083-92.

  30. Cilerdzic J, Stajic M, Vukojevic J, Duletic-Lausevic S. Oxidative stress and species of genus Ganoderma (higher Basidiomycetes). Int J Med Mushrooms. 2013;15(1):21-8.

  31. Roseiro LB, Rauter AP, Serralheiro ML. Polyphenols as acetylcholinesterase inhibitors: structural specificity and impact on human disease. Nutr Aging (Amst). 2012;1(2):99-111.

  32. Murray AP, Faraoni MB, Castro MJ, Alza NP, Cavallaro V. Natural AChE inhibitors from plants and their contribution to Alzheimer's disease therapy. Curr Neuropharmacol. 2013;11(4):388-413.

  33. Hajipour S, Sarkaki A, Farbood Y, Eidi A, Mortazavi P, Valizadeh Z. Effect of gallic acid on dementia type of Alzheimer disease in rats: electrophysiological and histological studies. Basic Clin Neurosci. 2016;7(2):97-106.

  34. Nagpal K, Singh SK, Mishra DN. Nanoparticle mediated brain targeted delivery of gallic acid: in vivo behavioral and biochemical studies for protection against scopolamine-induced amnesia. Drug Deliv. 2013;20(3-4):112-9.

Articles with similar content:

Investigations on Antioxidative Potential of Poroid Medicinal Mushroom Porodaedalea pini (Agaricomycetes)
International Journal of Medicinal Mushrooms, Vol.21, 2019, issue 6
Minni Singh, Niharika Kaushal, Navpreet Kaur, Gurpaul Singh Dhingra, Avneet Pal Singh
Evaluation of Bioactivities, Phenolic and Metal Content of Ten Wild Edible Mushrooms from Western Black Sea Region of Turkey
International Journal of Medicinal Mushrooms, Vol.21, 2019, issue 10
Demet Kizil, Semiha Yenigun, Tevfik Ozen, Ibrahim Turkekul, Hasan Cesur
Did the Iceman Know Better? Screening of the Medicinal Properties of the Birch Polypore Medicinal Mushroom, Piptoporus betulinus (Higher Basidiomycetes)
International Journal of Medicinal Mushrooms, Vol.17, 2015, issue 12
Maja Kozarski, Anita Klaus, Jovana Vunduk, Leonardus Johannes Lambertus Donatus Van Griensven, Predrag Petrovic, Miomir Niksic, Zeljko Zizak
Studies on the Triterpenoid Constituents of the Spores of Ganoderma lucidum (Curt.: Fr.) P. Karst. (Aphyllophoromycetideae)
International Journal of Medicinal Mushrooms, Vol.1, 1999, issue 2
De-Quan Yu, Ruo-Yun Chen
Protective Effect of Antioxidant Extracts from Grey Oyster Mushroom, Pleurotus pulmonarius (Agaricomycetes), Against Human Low-Density Lipoprotein Oxidation and Aortic Endothelial Cell Damage
International Journal of Medicinal Mushrooms, Vol.18, 2016, issue 2
Mohamad Hamdi Zainal Abidin, Noorlidah Abdullah, Nurhayati Zainal Abidin