Abstract
Ganoderma lucidum possesses a variety of valuable pharmacological activities, and it has long been used to prevent and treat various human diseases. Up to now, far too little attention has been paid to the liquid spawn of G. lucidum, and the development of the G. lucidum industry is constrained by them. This work aimed to study the key technologies and scale-up preparation of G. lucidum liquid spawn, to achieve large-scale preparation of liquid spawn and solve the problem of unstable quality of G. lucidum. The plate culture, primary shake flask culture, shake flask preparation, and fermentor preparation of G. lucidum liquid spawn were explored in the process of liquid fermentation. The results showed that plate broth volume significantly affected mycelial growth rate. Biomass in the primary shake flask culture is significantly influenced by the picking position of plate mycelium. An artificial neural network coupled with a genetic algorithm was used for carbon and nitrogen sources concentration optimization to increase biomass and substrate utilization. The optimized parameter combination is as follows: glucose, 14.5 g L−1; yeast extract powder, 8.5 g L−1. Under this condition, the biomass (9.82 g L−1) and biomass on reducing sugar (0.79 g g−1) increased by 18.03% and 27.41% compared to the control, respectively. The metabolic activity of liquid spawn prepared by different fermentation scales was diverse, and the liquid spawn prepared by the fermentor has better activity. Conceivably, the liquid spawn process can more conducive be applied to large-scale industrial production.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are included in this article.
References
Ahmad MF, Wahab S, Ahmad FA, Ashraf SA, Abullais SS, Saad HH (2022) Ganoderma lucidum: A potential pleiotropic approach of ganoderic acids in health reinforcement and factors influencing their production. Fungal Biol Rev 39:100–125. https://doi.org/10.1016/j.fbr.2021.12.003
Ahmad R, Riaz M, Khan A, Aljamea A, Algheryafi M, Sewaket D, Alqathama A (2021) Ganoderma lucidum (Reishi) an edible mushroom; a comprehensive and critical review of its nutritional, cosmeceutical, mycochemical, pharmacological, clinical, and toxicological properties. Phytother Res 35(11):6030–6062. https://doi.org/10.1002/ptr.7215
Atila F (2020) Comparative study on the mycelial growth and yield of Ganoderma lucidum (Curt.: Fr.) Karst. on different lignocellulosic wastes. Acta Ecol Sin 40(2):153–157. https://doi.org/10.1016/j.chnaes.2018.11.007
Abdullah N, Ismail R, Johari NMK, Annuar MSM (2013) Production of liquid spawn of an edible grey oyster mushroom, Pleurotus pulmonarius (Fr.) Quél by submerged fermentation and sporophore yield on rubber wood sawdust. Sci Hortic 161:65–69. https://doi.org/10.1016/j.scienta.2013.06.026
Asadi F, Barshan-Tashnizi M, Hatamian-Zarmi A, Davoodi-Dehaghani F, Ebrahimi-Hosseinzadeh B (2021) Enhancement of exopolysaccharide production from Ganoderma lucidum using a novel submerged volatile co-culture system. Fungal Biol 125(1):25–31. https://doi.org/10.1016/j.funbio.2020.09.010
Bishop KS, Kao CH, Xu Y, Glucina MP, Paterson RRM, Ferguson LR (2015) From 2000 years of Ganoderma lucidum to recent developments in nutraceuticals. Phytochemistry 114:56–65. https://doi.org/10.1016/j.phytochem.2015.02.015
Balamurugan JP, Supramani S, Usuldin SRA, Ilham Z, Klaus A, Ikram NKK, Wan WAAQI (2021) Efficient biomass-endopolysaccharide production from an identified wild-Serbian Ganoderma applanatum strain BGS6Ap mycelium in a controlled submerged fermentation. Biocatal Agric Biotechnol 37:102166. https://doi.org/10.1016/j.bcab.2021.102166
Cui J, Liu J, Chen X, Meng J, Wei S, Wu T, Zhang X (2022) Ganoderma Lucidum-derived erythrocyte-like sustainable materials. Carbon 196:70–77. https://doi.org/10.1016/j.carbon.2022.04.034
Ćilerdžić JL, Vukojević JB, Klaus AS, Ivanović ŽS, Blagojević JD, Stajić MM (2018) Wheat straw-a promising substrate for Ganoderma lucidum cultivation. Acta Sci Polonorum-Hortorum Cultus 17(1):13–22. https://doi.org/10.24326/asphc.2018.1.2
Ćilerdžić J, Vukojević J, Stajić M, Stanojković T, Glamočlija J (2014) Biological activity of Ganoderma lucidum basidiocarps cultivated on alternative and commercial substrate. J Ethnopharmacol 155(1):312–319. https://doi.org/10.1016/j.jep.2014.05.036
Chien YL, Ho CT, Chiang BH, Hwang LS (2011) Effect of fermentation time on antioxidative activities of Ganoderma lucidum broth using leguminous plants as part of the liquid fermentation medium. Food Chem 126(4):1586–1592. https://doi.org/10.1016/j.foodchem.2010.12.024
da Cruz MP, Felipini RB, Cardozo MM, Mazaro SM, Di Piero RM (2022) Ganoderma lucidum mycelial growth filtrate and the mycelial extract increase defense responses against Septoria leaf spot in tomato. Biol Control 173:105002. https://doi.org/10.1016/j.biocontrol.2022.105002
Du Z, Dong CH, Wang K, Yao YJ (2019) Classification, biological characteristics and cultivations of Ganoderma. Ganoderma Health. https://doi.org/10.1007/978-981-13-9867-4-2
Feng J, Feng N, Zhang JS, Yang Y, Jia W, Lin CC (2016) A new temperature control shifting strategy for enhanced triterpene production by Ganoderma lucidum G0119 based on submerged liquid fermentation. Appl Biochem Biotechnol 180(4):740–752. https://doi.org/10.1007/s12010-016-2129-1
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426–428. https://doi.org/10.1021/ac60147a030
Feng J, Feng N, Tang Q, Liu Y, Tang C, Zhou S, Lin CC (2021) Development and optimization of the triterpenoid and sterol production process with Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum strain G0017 (agaricomycetes), in liquid submerged fermentation at large scale. Int J Med Mushrooms 23(3):43–53
Hapuarachchi KK, Elkhateeb WA, Karunarathna SC, Cheng CR, Bandara AR, Kakumyan P, Wen TC (2018) Current status of global Ganoderma cultivation, products, industry and market. Mycosphere 9(5):1025–1052
Hou CT, Lin JT, Dulay RMR, Ray K (2017) Identification of molecular species of acylglycerols of Philippine wild edible mushroom, Ganoderma lucidum. Biocatal Agric Biotechnol 9:19–27. https://doi.org/10.1016/j.bcab.2016.10.013
Hsu KD, Cheng KC (2018) From nutraceutical to clinical trial: frontiers in Ganoderma development. Appl Microbiol Biotechnol 102(21):9037–9051. https://doi.org/10.1007/s00253-018-9326-5
Lu CT, Leong PY, Hou TY, Huang SJ, Hsiao YP, Ko JL (2018) Ganoderma immunomodulatory protein and chidamide down-regulate integrin-related signaling pathway result in migration inhibition and apoptosis induction. Phytomedicine 51:39–47. https://doi.org/10.1016/j.phymed.2018.06.023
Liang C, Tian D, Liu Y, Li H, Zhu J, Li M, Xia J (2019) Review of the molecular mechanisms of Ganoderma lucidum triterpenoids: Ganoderic acids A, C2, D, F, DM, X and Y. Eur J Med Chem 174:130–141. https://doi.org/10.1016/j.ejmech.2019.04.039
Lian J, Cheng R, Xiong L, Pang D, Tian X, Lei J, Zhu W (2019) Supercapacitors with high nitrogen content by cage-like Ganoderma lucidum spore. Appl Surf Sci 494:230–238. https://doi.org/10.1016/j.apsusc.2019.07.109
Lin Z (2019) Ganoderma (Lingzhi) in traditional Chinese medicine and Chinese culture. Ganoderma and Health: Biology, Chemistry and Ind. DOI: https://doi.org/10.1007/978-981-13-9867-4
Liu SR, Zhang WR, Kuang YB (2018) Production of stalk spawn of an edible mushroom (Pleurotus ostreatus) in liquid culture as a suitable substitute for stick spawn in mushroom cultivation. Sci Hortic 240:572–577. https://doi.org/10.1016/j.scienta.2018.06.068
Liu L, Feng J, Gao K, Zhou S, Yan M, Tang C, Zhang J (2022) Influence of carbon and nitrogen sources on structural features and immunomodulatory activity of exopolysaccharides from Ganoderma lucidum. Process Biochem 119:96–105. https://doi.org/10.1016/j.procbio.2022.05.016
Mohd Hanafiah Z, Wan Mohtar WHM, Abu Hasan H, Jensen HS, Klaus A, Wan-Mohtar WAAQI (2019) Performance of wild-Serbian Ganoderma lucidum mycelium in treating synthetic sewage loading using batch bioreactor. Sci Rep 9(1):1–12. https://doi.org/10.1038/s41598-019-52493-y
Ma L, Lin YQ, Yang C, Ying ZH, Jiang XL (2016) Production of liquid spawn of an edible mushroom, Sparassis latifolia by submerged fermentation and mycelial growth on pine wood sawdust. Sci Hortic 209:22–30. https://doi.org/10.1016/j.scienta.2016.06.001
Ren L, Zhang J, Zhang T (2021) Immunomodulatory activities of polysaccharides from Ganoderma on immune effector cells. Food Chem 340:127933. https://doi.org/10.1016/j.foodchem.2020.127933
Reis FS, Martins A, Vasconcelos MH, Morales P, Ferreira IC (2017) Functional foods based on extracts or compounds derived from mushrooms. Trends Food Sci Technol 66:48–62. https://doi.org/10.1016/j.tifs.2017.05.010
Strong PJ, Self R, Allikian K, Szewczyk E, Speight R, O’Hara I, Harrison MD (2022) Filamentous fungi for future functional food and feed. Curr Opin Biotechnol 76:102729. https://doi.org/10.1016/j.copbio.2022.102729
Sun B, You H, Xu JW (2021) Enhancement of ganoderic acid production by promoting sporulation in a liquid static culture of Ganoderma species. J Biotechnol 328:72–77. https://doi.org/10.1016/j.jbiotec.2021.01.014
Si J, Meng G, Wu Y, Ma HF, Cui BK, Dai YC (2019) Medium composition optimization, structural characterization, and antioxidant activity of exopolysaccharides from the medicinal mushroom Ganoderma lingzhi. Int J Biol Macromol 124:1186–1196. https://doi.org/10.1016/j.ijbiomac.2018.11.274
Tseng AJ, Tu TH, Hua WJ, Yeh H, Chen CJ, Lin ZH, Lin TY (2022) GMI, Ganoderma microsporum protein, suppresses cell mobility and increases temozolomide sensitivity through induction of Slug degradation in glioblastoma multiforme cells. Int J Biol Macromol 219:940–948. https://doi.org/10.1016/j.ijbiomac.2022.08.024
Wang T, Wang Y, Chen C, Ren A, Yu H, Zhao M (2021) Effect of the heme oxygenase gene on mycelial growth and polysaccharide synthesis in Ganoderma lucidum. J Basic Microbiol 61(3):253–264. https://doi.org/10.1002/jobm.202000622
Xu JW, Ji SL, Li HJ, Zhou JS, Duan YQ, Dang LZ, Mo MH (2015) Increased polysaccharide production and biosynthetic gene expressions in a submerged culture of Ganoderma lucidum by the overexpression of the homologous α-phosphoglucomutase gene. Bioprocess Biosyst Eng 38(2):399–405. https://doi.org/10.1007/s00449-014-1279-1
Yang M, Dai J, He M, Duan T, Yao W (2020) Biomass-derived carbon from Ganoderma lucidum spore as a promising anode material for rapid potassium-ion storage. J Colloid Interface Sci 567:256–263. https://doi.org/10.1016/j.jcis.2020.02.023
Yang X, Yang Y, Zhang Y, He J, Xie Y (2021) Enhanced exopolysaccharide production in submerged fermentation of Ganoderma lucidum by Tween 80 supplementation. Bioprocess Biosyst Eng 44(1):47–56. https://doi.org/10.1007/s00449-020-02418-1
Yang H, Min W, Bi P, Zhou H, Huang F (2013) Stimulatory effects of Coix lacryma-jobi oil on the mycelial growth and metabolites biosynthesis by the submerged culture of Ganoderma lucidum. Biochem Eng J 76:77–82. https://doi.org/10.1016/j.bej.2013.04.012
Zhang RY, Hu DD, Ma XT, Li SG, Gu JG, Hu QX (2014) Adopting stick spawn reduced the spawn running time and improved mushroom yield and biological efficiency of Pleurotus eryngii. Sci Hortic 175:156–159. https://doi.org/10.1016/j.scienta.2014.05.028
Zhou, X. W. (2017). Cultivation of Ganoderma lucidum. Edible and Medicinal Mushrooms: Technology and Applications, 385–413. https://doi.org/10.1002/9781119149446.ch18
Zeng P, Guo Z, Zeng X, Hao C, Zhang Y, Zhang M, Zhang L (2018) Chemical, biochemical, preclinical and clinical studies of Ganoderma lucidum polysaccharide as an approved drug for treating myopathy and other diseases in China. J Cell Mol Med 22(7):3278–3297. https://doi.org/10.1111/jcmm.13613
Funding
This work was supported by Agriculture Research System of Shanghai of China ([2022] 9), Project of Shanghai Academy of Agricultural and Sciences Excellent Team (2022A-03), Shanghai Academy of Agricultural Sciences Climbing Project.
Author information
Authors and Affiliations
Contributions
JF and JSZ conceptualized the idea of this study. JG perform the experiments and write the initial draft of the manuscript. YFL and CHT validate the data and provide overall supervision. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors have no relevant financial or non-financial interests to disclose.
Ethical approval
This article does not contain any studies with human participants or animals performed.
Consent to participate
Informed consent was obtained from all individual participants included in the study.
Consent to publish
The authors affirm that human research participants provided informed consent for publication.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Guo, J., Liu, YF., Tang, CH. et al. The key technologies of Ganoderma lucidum liquid spawn preparation and scale expansion. World J Microbiol Biotechnol 39, 138 (2023). https://doi.org/10.1007/s11274-023-03581-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-023-03581-9