Optimization of Extraction of Flavonoid, Total Phenolic, Antioxidant, and Antimicrobial Compounds from Ganoderma Lucidum by Maceration Method

Document Type : Research Article


Department of Food Science and Technology, Faculty of Agriculture, Varamin-Pishva Branch, Islamic Azad University, Varamin, I.R. IRAN


Plants are a rich source of phenolic and flavonoid compounds which are among the most important natural antioxidants. The aim of this study was to optimize the extraction of flavonoid, total phenolic, antioxidant, and antimicrobial compounds from Ganoderma (G) Lucidum by the maceration method. To do so, independent variables including temperature, extraction time, and type of solvent along with Box-Behnken Response Surface Methodology (RSM) were used. The results of single optimization of the independent variables showed the highest flavonoid content (15.19 mg/g) with 100% desirability at an extraction time of 48 h, the temperature of 60 °C by using ethanol solvent. The highest total phenolic content (16.96 mg/g) with 99.96% desirability was observed at an extraction time of 26 h, a temperature of 60 °C by using ethanol solvent. The highest amount of antioxidant compounds (3.03 mg/g) or the lowest IC50 value (mg/mL) with 100% desirability was found at an extraction time of 48 h at a temperature of 60 °C by using ethanol solvent. The results of simultaneous optimization of the extraction conditions by maceration method showed the highest flavonoid content (15.20 mg/g), total phenolic content (16.01 mg/g), and the lowest IC50 (3.03 mg/mL) with 98.157% desirability at extraction time of 48 h, the temperature of 60 °C using ethanol solvent. The highest mean Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of G. lucidum extract obtained by the maceration method were 2500 and 5000 μg/mL, respectively against Clostridium perfringens and Escherichia E. coli. The predicted optimized treatment had superior antimicrobial activity against Staphylococcus aureus with a non-growth halo (Zone of inhibition) diameter of 10.60 mm as compared to C. perfringens and E. coli. The results revealed that the G. lucidum extract obtained by the maceration method could be introduced as an antioxidant and antimicrobial source in marketable food products.


Main Subjects

[1] Mandal V., Mohan Y., Hemalatha S., Microwave Assisted Extraction ñ "An Innovative & Promising Extraction Tool for Medicinal Plant Research. Pharmaco. Rev., 1: 8-14 (2007).
[2] Kunle O.F., Eghareba H.O., Ahmadu P.O., Standardization of Herbal Medicines-A Review, Int. J. Biodivers Conserv., 4(3):101-112 (2012).
[3] Hayek S.A., Gyawali R., &Ibrahim S.A., Antimicrobial Natural Products. InA. M_endez-Vilas (Ed.), Microbial Pathogens and Strategies for Combating Them, Sci., Technol. and Edu., 2: 910-921 (2013).
[4] Wang L., Weller C.L., Recent Advances in Extraction of Nutraceuticals from Plants, Trends. Food Sci. Technol., 17: 300-312 (2006).
[7] Azmir J., Zaidul I.S.M., Rahman M.M., Sharif K.M., Mohamed A., Sahena F., Jahurul M.H.A., Ghafoor K., Norulaini N.A.N., Omar A.K.M., Techniques for Extraction of Bioactive Compounds from Plant Materials: A Review, J. Food Eng., 117: 426-436 (2013).
[8] Akinyele B.J., Obameso, J.O., Oladunmoye M.K., Phytochemical Screening and Antimicrobial Potentials of Three Indigenous Wild Ganoderma Mushrooms from ondo State Nigeria, Niger. J. Microb., 25: 2280-2290 (2011).
[9] Sardroodian M., Mehrban Sang Atash M., Aryanfar A., Optimization and Comparison of Effect of Different Extraction Conditions (Maceration and Ultrasound) on the Chemical Composition of Elaeagnus Angustifolia L. Extract, Quarterly of Echopytochem Medicinal Herbs., 4(3): 79-94 (2016).
[10] Izadi  Z., Sorooshzadeh A., Modarres Sanavi A.M., Asnaashari M., Aghaalikhani M., Davoodi P.A., Effect of Extraction Method on Antimicrobial Property of Aerial Parts of Echinacea Purpurea L. Against Some Pathogenic Bacteria, J Rafsanjan Faculty Medical Sci., 13: 267-80 (2014).
[11] Dashtimakan, A., Roodbari F., Mohajerani M., Mahmoodi Otaghvari A., Kavoosian S., Zahedi Zh., Hasanzadeh N., Evaluation of the Antiviral Effect of Oliveria Decumbens Vents in Controlling Herpes Simplex Virus Type1, J. Uni. Medical. Sci., 26(2): 143-125 (2018).
[12] Mayzumi F., Okamoto H., Mizuno T., Cultivation of Reishi (Ganoderma lucidum). Food Rev. Int., 13: 365 – 82 (1997).
[14] Badalyan S.M., Gharibyan N.G., Kocharyan A.E., Perspective in the Usage of Bioactive Substances of Medicinal Mushrooms in the Pharmaceutical and Cosmetic Industry, Intl. J. Medic. Mushrooms, 9(3,4): 275 (2007).
[15] Yang X., Chen C.h., Mi K., Yang Q., The Potential Use of Limulus G Test Assay for Evaluation of Immunomodalatory Activity of Ganoderma Polysaccarides, Int J Medici Mushrooms., 9 (3,4) :219-20 (2007).
[16] Vilkhu K., Mawson R., Simons L., Bates D. Applications and Opportunities for Ultrasound Assisted Extraction in the Food Industry, A Review, Innov. Food Sci. & Emerg. Technol., 9(2): 161-169 (2008).
[17] Ebrahimzadeh M.A., Pourmorad F., Hafezi S., Antioxidant Activities of Iranian Corn Silk, Turkish J. Bio., 32: 43-49 (2008).
[18] Chang C., Yang M., Wen H., Chern J., Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Analys., 10:178-182 (2002).
[19] Yao X., Zhang D., Zu Y., Fu Y., Luo M., Gu C., Li C.M.F., Efferth T., Free Radical Scavenging Capability, Antioxidant Activity and Chemical Constituents of Pyrola Incarnata Fisch. Leaves, Indust Crops and Product., 49: 247– 255 (2013).
[21] Mello B.C.B.S., Petrus J.C.C., Hubinger M.D., Concentration of Flavonoids and Phenolic Compounds in Aqueous and Ethanolic Propolis Extracts Through Nanofiltration. J. Food Eng., 96:533-539 (2010).
[22] Wang T.­Y., Li Q., Bi K.­S., Bioactive Flavonoids in Medicinal Plants: Structure, Activity and Biological Fate, Asian J. Pharma. Sci., 13(1): 12-23 (2018).
[23] Liazid A., Guerrero R.F., Cantos-Villar E., Microwave Assisted Extraction of Anthocyanins from Grape Skin, Food Chem., 124(3): 1238-1243 (2011).
[24] Mokhtarpour A., Extraction of Phenolic Compounds and Tannins from Pistachio By-products, Annual Res. & Rev. in Biology., 4(8): 1330-1338 (2014).
[25] Lee O. H., Lee B.Y., Lee J., Assessment of Phenolics-Enriched Extract and Fractions of Olive Leaves and their Antioxidant Activities, J. Bioresource Technol., 101(10): 3751-3754 (2009).
[26] Saltarelli R., Ceccaroli P.,  Lotti M., Zambonelli A., Buffalini  M., Casadei L., Vallorani L., Stocchi V., Biochemical characterisation and antioxidant activity of Mycelium of Ganoderma lucidum from Central Italy, Food Chem., 116: 143-51 (2009).
[27] Toprakçı İ., Kurtulbaş E., Pekel A. G., Şahin S.,  Application of D‐Optimal Design for Automatic Solvent Extraction of Carotenoid from Orange Peel. J Food Process and Preserv., e15724 (2021).
[28] Galanakis C.M., Goulas V., Tsakona S., Manganarisl G.A., Gekas V., A Knowledge Base for the Recovery of Natural Phenols with Different Solvents, Int. J. Food Proper., 16(2): 382-396 (2013).
[30] Al-Farsi M., Lee C.Y., Optimization of Phenolics and Dietary Fiber Extraction from Date Seeds, Food Chem., 108(3): 977-985 (2008).
[32] Ciğeroğlu Z., Aras Ö., Pinto C. A., Bayramoglu M., Kırbaşlar Ş. İ., Lorenzo J. M., Şahin S., Optimization of Ultrasound‐Assisted Extraction of Phenolic Compounds from Grapefruit (Citrus Paradisi Macf.) Leaves Via D‐Optimal Design and Artificial Neural Network Design with Categorical and Quantitative Variables, J. Sci. Food and Agri., 98(12): 4584-4596 (2018).
[33] Ghasemi K., Ghasemi Y., Ebrahimzadeh M.A., Antioxidant Activity, Phenol, and Flavonoid Contents of 13 Citrus Species Peek and Tissues, J. Pharmaceuti. Sci., 22(3): 277-281 (2009).
[35] Chitra K., Venkatesh R., Dhanalakshmi K., Sharavanan P.T., Bali Sasikumar C., Karthikeyani Vijayakumari K., Production and Economic Analysis of Oyster Mushroom (Pleurotus Florida), Int. J. Current Microb. and Applied. Sci., 7(90): 2319-7706 (2018).
[36] Mohammadi A., Ahmadzadeh Sani T., Kamali H., Alesheikh P., Fayzi P., Zarghami Moghaddam P., Evaluation of Phenolic Compounds and the Antibacterial Effect of Various Extracts of Aerial Parts of Scutellaria Pinnatifida on Typical Food-Born Pathogens, North khorasan Uni Medical Sci., 7(2): 393-403 (2015).
[37] Shanmugapriya K., Saravana P.S., Payal H., Peer Mohammed S., Antioxidant Potential of Pepper (Piper Nigrum Linn) Leaves and its Antimicrobial Potential Against Some Pathogenic Microbes, Indian J. Natu Pro And Resource., 3(4): 570-577 (2019).
[39] Mattila P., Hellstrom J., Phenolic Acids in Potatoes, Vegetables, and Some of Theirproducts. J. Food Composition Analysis., 20: 152 – 160 (2007).
[40] Natella   F. Nardini M., Di Filici M., Scaccini C., Benzoic Acid and Cinnamic Acid Derivatives As Antioxidants, Structure–Activity. J. Agri. and Food Chem., 47: 1453–1459 (1999).
[41] Andreasen M.F., Landbo A.K., Christensen  L.P., Hansen  Å., Meyer  A.S., Antioxidant Effects of Phenolic Rye (Secale Cereale L.) Extracts, Monomeric Hydroxycinnamates, and Ferulic Acid Dehydrodimers on Human Low-Density Lipoproteins, Journal of Agricultural and Food Chem., 49(8): 4090-4096 (2001).
[42] Koda T., Kuroda Y., Imai H., Protective Effect Ofrutin Against Spatial Memory Impairment Inducedby Trimethyltin in Rats. Nutr. Res., 28: 629–634 (2008).
[43] Sadeghi Mohoonak A.R, Salmanian Sh., Ghorbani M., Alami M., Evaluation of Antiradical and Antioxidant Activity and Determination of Phenolic Compounds of Crataegus Fruit, J. Iranian Food Indust. and Nutr. Sci., 8(1): 177-185 (2013).
[44] Vuorela S., Meyer A.S., Heinonen M., Impact of Isolation Method on Theantioxidant Activity of Rapeseed Meal Phenolics, J. Agri. and Food Chem., 52: 8202–8207 (2004).
[46] Heleno S.A., Barros L., Martins A., Queiroz M.J. R., Santos-Buelga C., Ferreira I.C., The Fruiting Body, Spores, and in Vitro Produced Mycelium of Ganoderma Lucidum from Northeast Portugal: A Comparative Study of the Antioxidant Potential of Phenolic and Polysaccharides Extracts, Food Res. Int., 46(1): 135-140. (2012).
[47] Dong Q., He D., Ni X., Zhou H., Yang  H., Comparative Study on Phenolic Compounds, Triterpenoids, and Antioxidant Activity of Ganoderma Lucidum Affected By Different Drying Methods, J. Food Measure Character., 13(4): 3198-3205 (2019).
[48] Costa T M., Kaufmann V.,  Paganelli C.J., Siebert D.A., Micke G. A., Alberton M. D., de Oliveira D., Kinetic Identification of Phenolic Compounds and Potential Production of Caffeic Acid by Ganoderma License in Solid-State Fermentation, Biopro. and Biosys. Eng., 42(8):1325-1332 (2019).
[49] Si J., Meng G., Wu Y., Ma H. F., Cui B.K., Dai Y.C., Medium Composition Optimization, Structural Characterization, and Antioxidant Activity of Exopolysaccharides from the Medicinal Mushroom Ganoderma Lingzhi, Int. J. Biology. Macromolecul., 124: 1186-1196 (2019).