Document Type : Research Article
Authors
Department of Microbiology, Faculty of Biological Sciences, NT.C., Islamic Azad University, Tehran, Iran
Abstract
Introduction
Probiotics are live, beneficial microorganisms that, when administered in adequate amounts, provide significant health benefits to the host. They play a vital role in maintaining and restoring gastrointestinal microbiota balance, improving digestion, modulating the immune system, and protecting against pathogenic bacteria. Prebiotics are non-digestible food components—typically fibers or complex carbohydrates—that selectively stimulate the growth and activity of beneficial gut microorganisms. By fostering a favorable environment for probiotics, prebiotics indirectly promote host health.
Recently, the search for natural and cost-effective prebiotic sources has intensified. Medicinal mushrooms, particularly Ganoderma lucidum (reishi) and Lentinula edodes (shiitake), have emerged as promising candidates due to their abundance of bioactive polysaccharides. These polysaccharides not only exhibit prebiotic potential but also possess antioxidant, immunomodulatory, and anti-inflammatory properties.
This study aimed to investigate the prebiotic properties of polysaccharides extracted from G. lucidum and L. edodes. Specifically, we evaluated their ability to stimulate the growth and metabolic activity of probiotic yeast and bacterial isolates obtained from traditional and industrial dairy products. These natural polysaccharides may contribute to the development of novel synbiotic products with enhanced health benefits.
Materials and Methods
Twenty dairy product samples were collected and subjected to microbial isolation to obtain yeast and bacterial strains with probiotic potential. Isolates were identified using morphological, biochemical, and molecular techniques, including PCR and sequencing.
Polysaccharides were extracted from the mycelium of G. lucidum and L. edodes via lyophilization and aqueous extraction. Carbohydrate content was determined using the phenol-sulfuric acid assay, while antioxidant capacity was assessed using the DPPH free radical scavenging method. FT-IR spectroscopy was employed to characterize chemical structures.
The prebiotic effects of the mushroom polysaccharides were evaluated by supplementing sugar-free culture media with these extracts. Growth and metabolic activity of selected probiotic isolates were compared to the ontrols supplemented with glucose and inulin. Enzymatic and acid digestion assays were also performed to assess the stability of the polysaccharides under simulated gastrointestinal conditions.
Results and Discussion
Multiple yeast and bacterial isolates were obtained from the dairy samples. Following biochemical screening, four isolates—A3, A10, B1, and B3—were selected for detailed analysis based on their probiotic potential. Antibiogram testing revealed varied resistance and sensitivity profiles.
Molecular identification showed that isolate A10 was closely related to Candida tropicalis, while A3 matched Saccharomyces cerevisiae. Among the bacterial isolates, B1 was identified as Lactobacillus casei and B3 as Lactobacillus acidophilus.
Polysaccharides extracted from G. lucidum and L. edodes exhibited lower reducing sugar content compared to inulin, indicating a more complex carbohydrate structure and greater resistance to enzymatic breakdown. Antioxidant assay demonstrated that, while inulin had the highest free radical scavenging activity, the mushroom polysaccharides also showed significant antioxidant properties.
FT-IR spectra confirmed the presence of characteristic polysaccharide functional groups, such as hydroxyl, carboxyl, and glycosidic linkages, consistent with previous findings on medicinal mushroom polysaccharides.
When added to sugar-free media, polysaccharides from both mushrooms stimulated the growth of probiotic isolates, though to a lesser extent than glucose and inulin. This suggests that while these fungal polysaccharides act as prebiotics, their fermentability and utilization by probiotics may differ from conventional prebiotics.
Optimal growth conditions for the probiotic isolates were observed at pH 5 and 37°C. The ability of these isolates to thrive under such conditions, along with their positive response to mushroom polysaccharides, underscores the potential of these compounds as functional prebiotic ingredients.
This study highlights the promising prebiotic and antioxidant properties of polysaccharides extracted from G. lucidum and L. edodes. These natural compounds supported the growth of beneficial probiotic yeasts and bacteria isolated from dairy products, suggesting their potential as synbiotic supplements.
Conclusion
Incorporating mushroom-derived polysaccharides into functional foods and dietary supplements could enhance gastrointestinal microbiota balance and promote overall health. However, further research involving in vivo animal models and human clinical trials is necessary to fully validate the health benefits and safety of these polysaccharides. Such studies will help clarify their mechanisms of action, determine optimal dosages, and assess long-term effects, paving the way for their application in health-promoting dietary and medicinal products.
Keywords
Main Subjects
©2025 The author(s). This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0)
- Alinia-Ahandani, E., Fazilati, M., Alizadeh, Z., & Boghozian, A. (2018). The introduction of some mushrooms as an effective source of medicines in Iran Northern. Biology and Medicine, 10(5), 1-5. https://doi.org/10.4172/0974-8369.1000451
- Appanna, V.D., & Appanna, V.D. (2018). Dysbiosis, probiotics, and prebiotics: in diseases and health. Human Microbes-The Power within: Health, Healing and Beyond, 81-122. https://doi.org/10.1007/978-981-10-7684-8_3
- Astashkina, A.P., Khudyakova, L., & Kolbysheva, Y.V. (2014). Microbiological quality control of probiotic products. Procedia Chemistry, 10, 74-79. https://doi.org/10.1016/j.proche.2014.10.014
- Baenas, I., Camacho‐Barcia, L., Miranda‐Olivos, R., Solé‐Morata, N., Misiolek, A., Jiménez‐Murcia, S., & Fernández‐Aranda, F. (2024). Probiotic and prebiotic interventions in eating disorders: A narrative review. European Eating Disorders Review, 32(6), 1085-1104. https://doi.org/10.1002/erv.3069
- Bisen, P., Baghel, R.K., Sanodiya, B.S., Thakur, G.S., & Prasad, G. (2010). Lentinus edodes: a macrofungus with pharmacological activities. Current Medicinal Chemistry, 17(22), 2419-2430. https://doi.org/10. 2174/092986710791698495
- Cappuccino, J.G., & Sherman, N. (2013). Microbiology: a laboratory manual: Pearson Higher Ed.
- Chen, Z., & Zhang, H. (2025). The interaction between Ganoderma lucidum polysaccharides and gut microbiota: Implications for immune health. Medicinal Plant Research, 15. https://doi.org/10.5376/mpr.2025.15.0001
- Chou, W.T., Sheih, I.C., & Fang, T.J. (2013). The applications of polysaccharides from various mushroom wastes as prebiotics in different systems. Journal of Food Science, 78(7), M1041-M1048. https://doi.org/10.1111/1750-3841.12160
- Cook, A., & Gilbert, R. (1969). The effect of sodium chloride on heat resistance and recovery of heated spores of Bacillus stearothermophilus. Journal of Applied Bacteriology, 32(1), 96-102. https://doi.org/10.1111/j.1365-2672.1969.tb02193.x
- Delzenne, N.M., & Bindels, L.B. (2015). Ganoderma lucidum, a new prebiotic agent to treat obesity? Nature Reviews Gastroenterology & Hepatology, 12(10), 553-554. https://doi.org/10.1038/nrgastro.2015.137
- Diosma, G., Romanin, D.E., Rey-Burusco, M.F., Londero, A., & Garrote, G.L. (2014). Yeasts from kefir grains: isolation, identification, and probiotic characterization. World Journal of Microbiology and Biotechnology, 30, 43-53. https://doi.org/10.1007/s11274-013-1419-9
- Dworkin, M., Falkow, S., Rosenberg, E., Schleifer, K.-H., & Stackebrandt, E. (2006). The prokaryotes: a handbook on the biology of bacteria (Vol. 1): Springer. https://doi.org/10.1007/0-387-30746-x
- Fan, H., Mazza, G., & Liao, X. (2010). Purification, composition and antioxidant activity of polysaccharides from wolfberry, cherry, kiwi and cranberry fruits. Croatian Journal of Food Science and Technology, 2(1), 9-17.
- Fontana, L., Bermudez-Brito, M., Plaza-Diaz, J., Munoz-Quezada, S., & Gil, A. (2013). Sources, isolation, characterisation and evaluation of probiotics. British Journal of Nutrition, 109(S2), S35-S50. https://doi.org/10.1017/S0007114512004011
- Gao, Y., Zhou, S., Chen, G., Dai, X., & Ye, J. (2002). A phase I/II study of a Ganoderma lucidum (Curt.: Fr.) P. Karst. Extract (Ganopofy) in patients with advanced cancer. International Journal of Medicinal Mushrooms, 4(3). https://doi.org/10.1615/IntJMedMushr.v4.i3.30
- Giavasis, I. (2014). Bioactive fungal polysaccharides as potential functional ingredients in food and nutraceuticals. Current Opinion in Biotechnology, 26, 162-173. https://doi.org/10.1016/j.copbio.2014.01.010
- Gomes, A.M., & Malcata, F.X. (1999). Bifidobacterium and Lactobacillus acidophilus: biological, biochemical, technological and therapeutical properties relevant for use as probiotics. Trends in Food Science & Technology, 10(4-5), 139-157. https://doi.org/10.1016/S0924-2244(99)00033-3
- Hearst, R., Nelson, D., McCollum, G., Millar, B.C., Maeda, Y., Goldsmith, C.E., & Moore, J.E. (2009). An examination of antibacterial and antifungal properties of constituents of Shiitake (Lentinula edodes) and Oyster (Pleurotus ostreatus) mushrooms. Complementary Therapies in Clinical Practice, 15(1), 5-7. https://doi.org/10.1016/j.ctcp.2008.10.002
- Holscher, H.D. (2017). Dietary fiber and prebiotics and the gastrointestinal microbiota. Gut Microbes, 8(2), 172-184. https://doi.org/10.1080/19490976.2017.1290756
- Huang, L.X., Gu, F.T., Zhu, Y.Y., Zhao, Z.C., Li, J.H., & Wu, J.Y. (2024). Bifidogenic properties of polysaccharides isolated from mushroom Lentinula edodes and enhanced immunostimulatory activities through Bifidobacterial fermentation. Food Bioscience, 62, 105121. https://doi.org/10.1016/j.fbio.2024.105121
- Huchchannanavar, S., Hasan, W., Kumar, D., & Gupta, S. (2024). Prebiotic potential of mushroom-derived compounds. In Mushroom Magic (pp. 228-243): CRC Press. https://doi.org/10.1201/9781003570257-14
- Jain, S.K., Jain, A., Gupta, Y., & Ahirwar, M. (2007). Design and development of hydrogel beads for targeted drug delivery to the colon. Aaps Pharmscitech, 8(3), 56. https://doi.org/10.1208/pt0803056
- Lindequist, U. (2024). Medicinal mushrooms as multicomponent mixtures—demonstrated with the example of Lentinula edodes. Journal of Fungi, 10(2), 153. https://doi.org/10.3390/jof10020153
- Madigan, M., Bender, K., Buckley, D., Sattley, W., & Stahl, D. (2018). Brock biology of microorganisms 15th edn, 71–74. In: Pearson Higher Ed.
- Manzanares, W., Lemieux, M., Langlois, P.L., & Wischmeyer, P.E. (2016). Probiotic and synbiotic therapy in critical illness: a systematic review and meta-analysis. Critical Care, 20, 1-19. https://doi.org/10.1186/s13054-016-1434-y
- Masuko, T., Minami, A., Iwasaki, N., Majima, T., Nishimura, S.-I., & Lee, Y.C. (2005). Carbohydrate analysis by a phenol–sulfuric acid method in microplate format. Analytical Biochemistry, 339(1), 69-72. https://doi.org/10.1016/j.ab.2004.12.001
- Mohanty, D., & Ray, P. (2016). Evaluation of probiotic and antimicrobial properties of lactobacillus strains isolated from dairy products. International Journal of Pharmacy and Pharmaceutical Sciences, 230-234. http://dx.doi.org/10.22159/ijpps.2016v8i11.12683
- Moradali, M.F., Hedjaroude, G.A., Mostafavi, H., Abbasi, M., Ghods, S., & Sharifi-Tehrani, A. (2007). The genus Ganoderma (Basidiomycota) in Iran. Mycotaxon, 99, 251-270. https://doi.org/10.1615/intjmedmushr.v7.i4.70
- Muszyńska, B., Pazdur, P., Lazur, J., & Sułkowska-Ziaja, K. (2017). Lentinula edodes (Shiitake)–biological activity. Medicina Internacia Revuo, 28(108), 189-195.
- Nielsen, S.S. (2009). Phenol-sulfuric acid method for total carbohydrates. In Food analysis laboratory manual (pp. 47-53): Springer. https://doi.org/10.1007/978-1-4419-1463-7_6
- Norajit, K., Kim, K.M., & Ryu, G.H. (2010). Comparative studies on the characterization and antioxidant properties of biodegradable alginate films containing ginseng extract. Journal of Food Engineering, 98(3), 377-384. https://doi.org/10.1016/j.jfoodeng.2010.01.015
- Pandey, K., Naik, S., & Vakil, B. (2015). Probiotics, prebiotics and synbiotics-a review. Journal of Food Science and Technology, 52(12), 7577-7587. https://doi.org/10.1007/s13197-015-1921-1
- Raghukumar, C. (2008). Marine fungal biotechnology: an ecological perspective. Fungal Diversity, 31, 19-35.
- Ren, G., Xu, L., Lu, T., & Yin, J. (2018). Structural characterization and antiviral activity of lentinan from Lentinus edodes mycelia against infectious hematopoietic necrosis virus. International Journal of Biological Macromolecules, 115, 1202-1210. https://doi.org/10.1016/j.ijbiomac.2018.04.132
- Roberfroid, M.B. (2000). Prebiotics and probiotics: are they functional foods? The American Journal of Clinical Nutrition, 71(6), 1682S-1687S. https://doi.org/10.1093/ajcn/71.6.1682S
- Rousta, N., Aslan, M., Yesilcimen Akbas, M., Ozcan, F., Sar, T., & Taherzadeh, M.J. (2024). Effects of fungal based bioactive compounds on human health. Critical Reviews in Food Science and Nutrition, 64(20), 7004-7027. https://doi.org/10.1080/10408398.2023.2178379
- Saad, N., Delattre, C., Urdaci, M., Schmitter, J.-M., & Bressollier, P. (2013). An overview of the last advances in probiotic and prebiotic field. LWT-Food Science and Technology, 50(1), 1-16. https://doi.org/10.1016/j.lwt.2012.05.014
- Stamets, P. (2011). Growing gourmet and medicinal mushrooms: Ten speed press.
- Synytsya, A., Míčková, K., Synytsya, A., Jablonský, I., Spěváček, J., Erban, V., & Čopíková, J. (2009). Glucans from fruit bodies of cultivated mushrooms Pleurotus ostreatus and Pleurotus eryngii: Structure and potential prebiotic activity. Carbohydrate Polymers, 76(4), 548-556. https://doi.org/10.1016/j.carbpol.2008.11.021
- Umu, Ö.C., Rudi, K., & Diep, D.B. (2017). Modulation of the gut microbiota by prebiotic fibres and bacteriocins. Microbial Ecology in Health and Disease, 28(1), 1348886. https://doi.org/10.1080/16512235.2017.1348886
- Wayne, P. (2010). Clinical and laboratory Standards Institute (CLSI)(2020) Performance standards for antimicrobial susceptibility testing: Background, Organization, Functions, and Processes. Journal Clinical Microbiology, 58(3), 01864-0186419. https://doi.org/10.1128/jcm.01864-19
- Xu, X., Yang, J., Ning, Z., & Zhang, X. (2015). Lentinula edodes-derived polysaccharide rejuvenates mice in terms of immune responses and gut microbiota. Food & Function, 6(8), 2653-2663. https://doi.org/10.1039/C5FO00689A
- Yang, K., Zhang, Y., Cai, M., Guan, R., Neng, J., Pi, X., & Sun, P. (2020). In vitro prebiotic activities of oligosaccharides from the by-products in Ganoderma lucidum spore polysaccharide extraction. RSC Advances, 10(25), 14794-14802. https://doi.org/10.1039/C9RA10798C
- Yang, L., Kang, X., Dong, W., Wang, L., Liu, S., Zhong, X., & Liu, D. (2022). Prebiotic properties of Ganoderma lucidum polysaccharides with special enrichment of Bacteroides ovatus and uniformis in vitro. Journal of Functional Foods, 92, 105069. https://doi.org/10.1016/j.jff.2022.105069
- Yin, C., Li, C., Ma, K., Fan, X., Yao, F., Shi, D., & Gao, H. (2023). The physicochemical, antioxidant, hypoglycemic and prebiotic properties of γ-irradiated polysaccharides extracted from Lentinula edodes. Food Science and Biotechnology, 32(7), 987-996. https://doi.org/10.1007/s10068-022-01234-5
- Zarmi, A.H., Tasharofi, Z., Alvandi, H., Barshan-tashnizi, M., Ebrahimi-Hosseinzadeh, B., & Hosseini, Z.B.M. (2022). A kinetic modeling of growth and mycelial exopolysaccharide production by Lentinus edodes (Shiitake edible mushroom): Enhancemet of production in volatile co-cultures for shiitake edible mushroom. Applied Food Biotechnology, 9(1), 67-78. https://doi.org/10.17474/artvinofd.953998
- Zhao, J., Zhao, J., Zang, J., Peng, C., Li, Z., & Zhang, P. (2024). Isolation, identification, and evaluation of lactic acid bacteria with probiotic potential from traditional fermented sour meat. Frontiers in Microbiology, 15, 1421285. https://doi.org/10.3389/fmicb.2024.1421285
- Zhao, L., Dong, Y., Chen, G., & Hu, Q. (2010). Extraction, purification, characterization and antitumor activity of polysaccharides from Ganoderma lucidum. Carbohydrate Polymers, 80(3), 783-789. https://doi.org/10.1016/j.carbpol.2009.12.029
- Zheng, M., Pi, X., Li, H., Cheng, S., Su, Y., Zhang, Y., & Jiang, Y. (2024). Ganoderma polysaccharides are potential prebiotics: a review. Critical Reviews in Food Science and Nutrition, 64(4), 909-927. https://doi.org/10.1080/10408398.2022.2110035
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