Research Report

Industrial-Scale Cultivation of Ganoderma lucidum: Challenges and Technological Innovations  

Jianfeng Wu1 , Ting Wu2 , Jinrong Chen3 , Haijuan Zhang4
1 Lishui Crop Research Institute, Lishui, 323000, Zhejiang, China
2 Longquan Xianzhitang Biological Technology Co., Ltd., Longquan, 323700, Zhejiang, China
3 Zhejiang Yuanjian Biopharmaceutical Co., Ltd., Longquan, 323700, Zhejiang, China
4 Longquan Agricultural and Rural Bureau, Longquan, 323700, Zhejiang, China
Author    Correspondence author
Medicinal Plant Research, 2024, Vol. 14, No. 6   doi: 10.5376/mpr.2024.14.0027
Received: 08 Oct., 2024    Accepted: 13 Nov., 2024    Published: 28 Nov., 2024
© 2024 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Wu J.F., Wu T., Chen J.R., and Zhang H.J., 2024, Industrial-scale cultivation of Ganoderma lucidum: challenges and technological innovations, Medicinal Plant Research, 14(6): 320-333 (doi: 10.5376/mpr.2024.14.0027)

Abstract

Ganoderma lucidum is highly valued for its rich bioactive compounds, offering a range of therapeutic effects, including immune enhancement, antioxidant, anti-inflammatory, etc. With the growing global demand for G. lucidum products, industrial-scale cultivation has become increasingly important. This study comprehensively explores the industrial-scale cultivation of G. lucidum, with a focus on analyzing its challenges and corresponding solutions. In response to key issues such as environmental control and contamination management in the cultivation process, innovative methods such as substrate optimization and the introduction of bioreactor technology are proposed to improve the production efficiency, product quality, and sustainability of G. lucidum cultivation. The study demonstrates that industrial-scale cultivation not only reduces costs and increases yield but also ensures the efficacy and quality standards of G. lucidum products. Continued development and promotion of large-scale cultivation techniques are crucial to meeting global market demand, while providing valuable reference and guidance for the application of G. lucidum in pharmaceuticals and health foods.

Keywords
Ganoderma lucidum; Large-scale cultivation; Technological innovation; Bioreactor; Substrate optimization; Sustainable strategies

1 Introduction

Ganoderma lucidum, commonly known as Reishi or Lingzhi, is a medicinal mushroom highly valued for its bioactive compounds, which have been widely used in traditional medicine, particularly in East Asia (El Sheikha, 2022). This fungus has garnered significant global interest due to its numerous therapeutic properties, including immune enhancement, anti-inflammatory, antioxidant, and anti-tumor effects (Lu et al., 2020; Ahmad et al., 2021). As the demand for Ganoderma lucidum-based products continues to rise, efforts to optimize and expand its cultivation have become increasingly critical. The shift towards industrial-scale production is essential to meet global demand and enhance accessibility, thereby promoting further research and applications of this versatile mushroom. 

 

Ganoderma lucidum is traditionally cultivated using several methods, including log cultivation, sawdust bag cultivation, and submerged fermentation. Each method has its own advantages and challenges, which influence the yield, quality, and production cost of Ganoderma lucidum. Log cultivation, for instance, simulates natural growing conditions, allowing the mushroom to develop its full range of bioactive compounds; however, it requires a long growth cycle and extensive land resources (Geetha et al., 2012). In contrast, sawdust bag cultivation offers higher control over the growth environment, resulting in faster yields, but it requires significant labor and careful environmental management (Sudheer et al., 2018; Chanshorphea, 2019). Submerged fermentation, often used for extracting specific metabolites, provides a more consistent and efficient production, yet requires sophisticated equipment and expertise (Feng et al., 2021). As the industry grows, developing and adopting innovative cultivation techniques that balance cost-efficiency, quality, and scalability are of paramount importance.

 

The growing interest in the health-promoting properties of Ganoderma lucidum has led to a surge in demand for its products, necessitating the transition from small-scale to industrial-scale production. Industrial cultivation offers several significant benefits, including the ability to ensure a consistent supply of high-quality raw materials, reduce production costs, and implement standardization of cultivation protocols (Wagner et al., 2003; Sudheer et al., 2018). Moreover, industrial-scale production enables the exploration of new applications in pharmaceuticals, nutraceuticals, and functional foods, expanding the market potential of Ganoderma lucidum (Ahmad et al., 2021). By leveraging large-scale production techniques, it becomes feasible to optimize the extraction of bioactive compounds, ensuring that Ganoderma lucidum-based products retain their efficacy and meet regulatory standards. The transition to industrial production is also crucial for achieving sustainable practices, minimizing resource usage, and ensuring economic viability for growers and manufacturers alike.

 

This study explores the challenges and technological innovations involved in the industrial cultivation of Ganoderma lucidum, identifying key obstacles that hinder efficient large-scale production, such as environmental control, contamination risks, and resource management. The study focuses on analyzing the technological innovations that address these challenges, including substrate optimization, advancements in automated cultivation systems, and bioreactor technologies. It proposes strategies to enhance production efficiency, product quality, and sustainability of Ganoderma lucidum. This study aims to promote the widespread adoption of best practices for large-scale industrial cultivation of Ganoderma lucidum and to improve the global availability of Ganoderma-based health products.

 

2 Biological and Ecological Requirements of Ganoderma lucidum

2.1 Growth conditions and nutrient requirements

Ganoderma lucidum requires a variety of nutrients to thrive. The primary nutrients include carbon, nitrogen, and essential minerals. The carbon source is typically derived from lignocellulosic materials such as wood, which provides the necessary energy for growth and development. The study found that Ganoderma lucidum can effectively utilize various lignocellulosic waste materials as substrates to produce cellulase and xylanase through solid-state fermentation. In particular, lignocellulosic waste from the Amazon region, such as marupa wood chips and açaí seeds, significantly enhances the enzyme activity of Ganoderma lucidum under appropriate supplementation conditions, demonstrating high application potential (de Oliveira Júnior et al., 2022). Nitrogen is another critical nutrient, often supplied through organic or inorganic means, which supports protein synthesis and other metabolic processes. Additionally, essential minerals such as potassium, phosphorus, and magnesium play vital roles in enzymatic functions and structural integrity of the fungal cells (Swallah et al., 2023).

 

Trace elements, including iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu), significantly influence the growth and development of Ganoderma lucidum. These elements are involved in various biochemical pathways and enzymatic reactions. For example, Mn2+ significantly impact the growth and metabolism of G. lucidum by promoting the synthesis of manganese peroxidase. Studies have shown that an increase in manganese significantly enhances the production of polysaccharides, triterpenoids, and the total manganese content in G. lucidum. Experimental results also indicate that manganese ions can regulate metabolite levels in G. lucidum, particularly by significantly upregulating certain metabolites, such as aromatic alcohols and palmitoylethanolamide (Zhang et al., 2019). Additionally, copper ions influence the growth of G. lucidum mycelium and the biosynthesis of ganoderic acid by increasing intracellular ROS levels. Furthermore, research has shown that the elevation of intracellular calcium ion levels can reduce ROS by activating antioxidant enzymes, thereby providing feedback regulation of mycelial growth. This indicates that copper ions affect the developmental process of G. lucidum through the interaction between ROS and calcium ions (Gao et al., 2018). Studies have shown that the presence and concentration of these trace elements can alter the microbial community structure in the soil, which in turn affects the growth conditions for G. lucidum (Ren et al., 2020).

 

2.2 Environmental factors influencing yield and quality

Temperature and humidity are critical environmental factors that directly impact the yield of Ganoderma lucidum. The study by Huynh and To (2023) demonstrated that G. lucidum grows best at temperatures of 25 °C to 30 °C and humidity levels of 60% to 70%, under which the highest polysaccharide content is produced. This indicates that temperature and humidity have a direct impact on the production of active compounds in G. lucidum. Nguyen et al. (2019) explored the cultivation conditions of the G. lucidum GA3 strain, and their results also showed that at temperatures between 25 °C and 30 °C, combined with an appropriate substrate, optimal fruiting body development and biomass yield could be achieved.

 

Light and ventilation are essential for the quality of G. lucidum. While G. lucidum can grow in low-light conditions, exposure to light is necessary for the development of its characteristic color and bioactive compounds. Proper ventilation is equally important as it helps in regulating the levels of carbon dioxide and oxygen, which are crucial for respiration and metabolic activities. A study has shown that by adjusting lighting and ventilation conditions, the content of bioactive compounds (such as phenolics, flavonoids, water-soluble polysaccharides, and ganoderic acids) in antler-shaped G. lucidum fruiting bodies can be significantly increased. This morphology of G. lucidum develops under limited ventilation, leading to an increase in carbon dioxide concentration, which promotes the synthesis of its bioactive compounds (Sudheer et al., 2018). Poor ventilation may cause carbon dioxide accumulation, negatively impacting the growth and quality of G. lucidum (Swallah et al., 2023). Another study explored the effects of different wavelengths of LED light (red, blue, green) on G. lucidum mycelial growth and antioxidant activity. The results indicated that mycelial growth was fastest under red light, while antioxidant activity was highest under blue light, and total phenolic content significantly increased under red light. This suggests that lighting not only affects the growth rate of G. lucidum but also has a significant impact on its antioxidant capacity (Alcazar et al., 2021).

 

2.3 Comparison of wild versus cultivated Ganoderma lucidum

Wild Ganoderma lucidum typically grows on decaying hardwood trees in forested areas. The natural environment provides a diverse range of nutrients and ecological interactions that contribute to its growth. Wild G. lucidum is often considered to have a richer profile of bioactive compounds due to the varied and complex interactions with other organisms and environmental factors. However, the yield and consistency of wild G. lucidum can be unpredictable due to the variability in natural conditions (Ren et al., 2020).

 

Cultivating Ganoderma lucidum offers several advantages, including controlled growth conditions, consistent yield, and the ability to optimize the production of specific bioactive compounds. Cultivation allows for the manipulation of nutrient availability, environmental conditions, and other factors to maximize growth and quality. However, challenges remain, such as the need for precise control of environmental parameters and the potential impact on soil and microbial communities. Continuous cultivation can alter soil properties and microbial diversity, which may affect long-term sustainability and soil health (Ren et al., 2020).

 

3 Industrial Cultivation Techniques of Ganoderma lucidum

3.1 Traditional cultivation methods

Log-based cultivation is one of the oldest and most traditional methods for cultivating Ganoderma lucidum. This technique involves inoculating wood logs with G. lucidum spawn and allowing the fungus to colonize the wood over several months. The logs are typically made from hardwood species such as poplar, oak, or other suitable trees. For example, the study by Bijalwan et al. (2021) examined the growth performance of G. lucidum cultivated in the Garhwal Himalayas of India. The research revealed that the growth cycle of G. lucidum on poplar logs varied depending on temperature differences, with higher temperatures shortening the growth cycle. The study indicated that warmer climates facilitate the early development of G. lucidum, but yield gradually decreased during the subsequent fruiting period. Qiu et al. (2023), using Fourier-transform infrared spectroscopy, studied the impact of different tree species on the cultivation of G. lucidum and found that broadleaf species such as Qv and Ps influenced the quality of G. lucidum fruiting bodies, especially the differences in protein content. This study provides a scientific basis for selecting the optimal tree species for G. lucidum cultivation. This method is widely used due to its simplicity and the natural environment it provides for the G. lucidum. However, it requires a longer cultivation period and is susceptible to contamination by other fungi (Tong et al., 2020; Bijalwan et al., 2021).

 

Sawdust cultivation is another traditional method that is popular due to its high efficiency and shorter cultivation cycle. In this system, sawdust is mixed with other organic materials, packed into plastic bags or bottles, and then inoculated with G. lucidum spores. Thiribhuvanamala and Krishnamoorthy (2021) found that using rubber tree sawdust as a substrate resulted in a biological efficiency of 44.3%, with earlier fruiting body formation and harvest periods demonstrating clear advantages of this method. Additionally, the addition of nitrogen salts and sodium phosphate to the sawdust substrate significantly enhanced the growth rate of G. lucidum mycelium, greatly reducing the time required to fully colonize the substrate. Notably, in substrates supplemented with nitrogen salts, the growth rate of G. lucidum reached up to 3.22 mm per day (Fedorov et al., 2022). This method has been successfully used with various types of sawdust, including those from pine and other hardwoods (Adongbede and Atoyebi, 2021; Fedorov et al., 2022; Oke et al., 2022).

 

3.2 Controlled environment systems

Greenhouse cultivation offers a controlled environment that can optimize the growth conditions for G. lucidum. By regulating temperature, humidity, and light, greenhouses can create an ideal setting for the fungus to thrive. In the continuous cultivation of Ganoderma lucidum, it was found that antagonistic fungi in the soil can affect its growth. The study showed that by applying a water immersion treatment, the number of antagonistic fungi was significantly reduced, thereby increasing the yield and spore production of Ganoderma lucidum (Tong et al., 2020). This method allows for year-round cultivation and can significantly increase yield and quality. Greenhouses also provide protection from environmental contaminants and pests, which can be a major issue in traditional outdoor cultivation methods (Bijalwan et al., 2021).

 

Technological advancements have driven the development of automated environmental control systems, which can precisely manage the conditions within G. lucidum cultivation facilities. For instance, parameters such as temperature, humidity, CO2 concentration, and light intensity can be adjusted through sensors and computer algorithms. These systems have shown significant effectiveness in both fully and partially automated modes, substantially improving crop quality and yield in greenhouses (Pryahin et al., 2022). Juleang and Mitath (2021) found that using wireless sensor systems to collect real-time data, combined with predictive algorithms for environmental control, greatly enhances the precision of crop environment management. For example, the use of such systems can significantly improve the accuracy of controlling soil moisture and air humidity, which is particularly important for fungi like G. lucidum that rely on stable humidity conditions.

 

3.3 Advances in substrate preparation

Recent innovations in substrate preparation have focused on the use of both organic and synthetic materials to improve the growth and yield of G. lucidum. Research has shown that G. lucidum can grow on various lignocellulosic substrates, and its productivity can be enhanced by adding supplements such as olive oil or copper. These substrates influence the active chemical components of G. lucidum, such as total triterpenes, ganoderic acids, and phenolic compounds, thus altering the composition of its bioactive substances (Bidegain et al., 2019). Pine sawdust, after pretreatment with a water jet disperser, can be used as a substrate for G. lucidum cultivation. The addition of nitrogen salts such as ammonium nitrate and monoammonium phosphate significantly accelerates the growth of G. lucidum, reducing the time needed to fully colonize the substrate and increasing the content of bioactive substances (Fedorov et al., 2022). 

 

Another study compared the effects of different lignocellulosic wastes as substrates for G. lucidum cultivation. The results showed that traditional substrates, oak (OS) and poplar (PS) sawdust, performed best in terms of G. lucidum yield and biological efficiency. Oak substrate produced a yield of 86.1 g/kg with a biological efficiency of 24.7%. In contrast, cottonseed meal (CSM) performed the worst as a substrate, with a yield of only 28.6 g/kg and a biological efficiency of 8.9% (Figure 1). The experiment also found that the growth rate of mycelium was negatively correlated with the cellulose and hemicellulose content of the substrate, while G. lucidum yield was positively correlated with the cellulose and lignin content of the substrate. This indicates that G. lucidum prefers substrates rich in cellulose and lignin (Atila, 2020). Additionally, effective substrate sterilization is crucial for preventing contamination and ensuring successful G. lucidum cultivation. Common sterilization techniques include autoclaving, pasteurization, and chemical sterilization, which help eliminate harmful microorganisms from the substrate. These methods contribute to creating an optimal environment for G. lucidum, resulting in better yields and higher quality mushrooms (Adongbede and Atoyebi, 2021).

 

Figure 1 Ganoderma lucidum fruit bodies grown on substrates tested in the study (Adopted from Atila, 2020)

Image caption: The figure clearly shows that G. lucidum fruit bodies grown on substrates of oak sawdust (OS) and poplar sawdust (PS) exhibit superior diameter, thickness, and morphology compared to other substrates, with larger pileus diameters and thicker stipes. In contrast, the fruit bodies grown on wheat straw (WS), sunflower meal (SFM), cottonseed meal (CSM), soybean straw (SBS), and bean straw (BS) substrates are smaller and thinner. The results indicate that substrates with high cellulose and lignin content can effectively promote the growth and yield of G. lucidum (Adapted from Atila, 2020)

 

4 Challenges in Industrial-Scale Production

4.1 Contamination and pest management

One of the primary challenges in the industrial-scale cultivation of Ganoderma lucidum is the management of contamination by other fungi and bacteria. Studies have shown that non-Ganoderma fungi such as Trichoderma and Mucor can significantly hinder the growth and fruiting body development of G. lucidum. These antagonistic fungi proliferate in nutrient-rich soils, leading to substantial cultivation problems (Tong et al., 2020). Additionally, the microbial community structure of the soil, wood segments, and tree roots can be significantly altered by continuous cultivation of G. lucidum, which may further complicate contamination control (Ren et al., 2020).

 

Effective pest management strategies are crucial for maintaining the health and productivity of G. lucidum cultures. One promising approach involves the use of ultraviolet ray irradiation to develop strains with enhanced resistance to microbial invasions. For instance,Through ultraviolet (UV) radiation, mutant strains of G. lucidum were generated, and it was found that the mutant strain UV119 had significantly higher yields compared to the control strain. This indicates that genetic variation among strains plays a crucial role in stabilizing and improving G. lucidum yield. Additionally, the mutant strain exhibited stronger resistance to microbial invasion, which could help mitigate the negative effects of soil microorganisms on yield (Tang et al., 2023). Additionally, sanitation methods such as waterlogging have been found to significantly reduce the number of contaminant colonies, thereby improving the yield and quality of G. lucidum (Tong et al., 2020).

 

4.2 Yield variability and consistency

Yield variability in G. lucidum cultivation can be attributed to several factors, including genetic differences among strains, environmental conditions, and soil properties. Significant changes in the microbial community structure of the soil, logs, and tree roots were observed before and after G. lucidum cultivation, particularly in the composition of fungal phyla (Ascomycota and Basidiomycota) and bacterial phyla (Proteobacteria and Actinobacteria). The study also revealed that soil properties, such as organic matter content, pH, nitrogen, and carbon levels, underwent changes after cultivation, and these changes were closely related to alterations in the microbial community structure (Ren et al., 2020). Moreover, the scarcity of natural resources and the strict growth conditions required for G. lucidum further contribute to yield inconsistencies (Tang et al., 2023).

 

To improve yield consistency of G. lucidum, it is essential to adopt advanced strain breeding techniques and optimize cultivation conditions. Optimizing soil properties and microbial community structure through controlled fermentation and the use of agro-industrial by-products as feed can promote yield stability (Kachrimanidou et al., 2023). In a large-scale liquid fermentation experiment, optimizing air supply strategies significantly increased the production of triterpenes and sterols in G. lucidum strain G0017, reaching 3.34 and 3.46 g/L, respectively. This optimization process holds potential for application in industrial-scale production (Feng et al., 2021). Hu et al. (2018) explored the effects of different nitrogen sources, carbon sources, and air supply on ganoderic acid accumulation and designed a bioreactor suitable for liquid static fermentation. The total yield of the top five ganoderic acids reached 963 mg/L, demonstrating the potential of optimized cultivation conditions for the production of G. lucidum triterpenoids.

 

4.3 Economic and logistical barriers

The industrial-scale production of G. lucidum requires substantial initial investment and ongoing operating costs. These costs are associated with the need for specialized cultivation facilities, advanced pest management systems, and high-quality substrates. The economic viability of G. lucidum cultivation is further challenged by the need for continuous monitoring and optimization of growth conditions to ensure high yields and quality (Tang et al., 2023).

 

Supply chain and distribution challenges also pose significant barriers to the large-scale production of G. lucidum. The perishable nature of the mushroom and its products necessitates efficient logistics and storage solutions to maintain quality from farm to market. Additionally, the variability in yield and quality can complicate supply chain management, making it difficult to meet market demands consistently (Kachrimanidou et al., 2023; Tang et al., 2023). Effective strategies to address these challenges include the development of robust supply chain networks and the use of bioprocessing techniques to extend the shelf life of G. lucidumproducts (Kachrimanidou et al., 2023).

 

5 Technological Innovations

5.1 Genetic improvements and strain selection

Recent advancements in the genetic improvement of G. lucidum have focused on cultivating strains with higher yields of bioactive compounds. Whole-genome sequencing and transcriptomic analysis have revealed that Tween80 enhances the production of extracellular polysaccharides in G. lucidum. Differential gene expression analysis identified multiple metabolic pathways involved in polysaccharide synthesis, providing a theoretical basis for breeding G. lucidum strains with higher polysaccharide content (Wu et al., 2022). One study used UV irradiation-induced mutagenesis to develop the G. lucidum mutant strain UV119, which showed an 8.67% increase in spore powder yield compared to its parent strain, along with enhanced resistance to microbial invasion. This technique has proven to be an effective method for improving G. lucidum strains, leading to higher yield and quality of spore powder (Tang et al., 2023).

 

Another study demonstrated that introducing the endogenous glyceraldehyde-3-phosphate dehydrogenase (gpd) intron 1 into G. lucidum significantly enhances the expression of exogenous genes, thereby improving its molecular breeding capacity. This offers a new approach to regulating G. lucidum metabolism and increasing the yield of bioactive compounds (You et al., 2021).

 

5.2 Metabolic engineering for improved bioactive compound production

Metabolic engineering involves the modification of metabolic pathways to increase the production of specific compounds. In Ganoderma lucidum, pathway modifications can be implemented to enhance the synthesis of triterpenoids, which are key bioactive compounds with significant medicinal properties. Optimizing culture conditions and genetic pathways can lead to higher yields of these valuable compounds (Hu et al., 2018).

 

Polysaccharides are another group of important bioactive compounds produced by Ganoderma lucidum. Through metabolic engineering, it is possible to increase the production of these compounds by optimizing the carbon and nitrogen sources in the cultivation medium. This approach has shown significant improvements in both volumetric and specific yields of polysaccharides, making it a viable alternative to traditional cultivation methods (Alsaheb et al., 2020).

 

5.3 Bioreactor technology for Ganoderma lucidum

Bioreactor technology offers several advantages for the cultivation of G. lucidum mycelia. Submerged fermentation in bioreactors allows for better control over the growth environment, leading to higher yields of bioactive compounds. For example, a study demonstrated that using a 16-L stirred tank bioreactor under controlled pH conditions significantly increased the production of exopolysaccharides (EPS) (Alsaheb et al., 2020). This method is more efficient than traditional solid-state fermentation, providing a scalable solution for industrial applications.

 

Scaling up production using bioreactor technologies has shown promising results. The optimization of carbon and nitrogen sources in a bioreactor setup led to a substantial increase in both volumetric and specific yields of EPS (Alsaheb et al., 2020). Additionally, the use of agro-industrial by-products as feedstocks in bioreactors has been explored, demonstrating the potential for sustainable and cost-effective production of G. lucidum bioactive compounds (Kachrimanidou et al., 2023). This approach not only enhances productivity but also aligns with the principles of bioeconomy by utilizing waste materials.

 

Case Studies

6.1 Large-scale efficient breeding of the Ganoderma lucidum UV119 mutant strain

The spore powder yield of G. lucidum is limited and prone to microbial contamination, significantly affecting the efficiency and stability of large-scale production. To increase spore powder yield and enhance resistance to microbial invasion, researchers employed ultraviolet mutagenesis technology to develop the G. lucidum UV119 mutant strain (Tang et al., 2023). This strain exhibits significantly higher spore yield and stronger resistance to microbial invasion. Compared to the original strain G0109, UV119's spore yield increased by 19.27%, fruiting body yield increased by 8.67%, and its antimicrobial ability surpassed that of the current main variety "Longzhi No.1" (Figure 2).

 

Figure 2 Production of spores during the strain cultivation. (A): Spore collection; (B): LZ-1; (C): mutagenic strain UV119 (Adopted from Tang et al., 2023)

Image caption: The figure shows the spore production of the Ganoderma lucidum UV119 mutant strain and "Longzhi No.1" over two growth cycles. A shows the spore powder collection process of the UV119 mutant strain, while B and C compare the production of the "Longzhi No.1" and UV119 strains, respectively. The experimental results indicate that UV119's spore yield in the first growth cycle was 26.67% higher than that of "Longzhi No.1," and in the second growth cycle, its spore yield was nearly 10 times that of "Longzhi No.1." This figure reveals the high-yield capacity of the UV119 strain across multiple growth cycles, confirming its significant spore yield advantage over "Longzhi No.1" (Adapted from Tang et al., 2023)

 

The results indicate that ultraviolet mutagenesis is an efficient strain improvement method, helping to enhance the yield and quality of G. lucidum spore powder, providing strong technical support for the development of the G. lucidum industry. The high-yield capability of the UV119 strain across multiple growth cycles demonstrates its efficiency in practical applications, with promising prospects for large-scale adoption.

 

6.2 Optimization of large-scale liquid fermentation process for triterpenoid and sterol production from G. lucidum

G. lucidum, a traditional medicinal fungus in China, is rich in triterpenoids and sterols, which have various bioactive properties such as antitumor, hepatoprotective, and antihypertensive effects (Zhao et al., 2019). However, traditional cultivation methods have low yields and cannot meet the demands of large-scale production. Liquid submerged fermentation, with its shorter production cycle, high controllability, and large output, has become an ideal method for large-scale production of G. lucidum.

 

Feng et al. (2021) conducted an optimization study on large-scale production using liquid submerged fermentation of the G. lucidum G0017 strain. By adjusting different aeration rates, they significantly increased the yield of triterpenoids and sterols. The experiment showed that an aeration rate of 1.25 vvm during the early fermentation stage effectively promoted mycelial growth, while adjusting to 1.5 vvm in the later stage greatly enhanced triterpenoid and sterol synthesis. The optimized process achieved triterpenoid and sterol yields of 3.34 g/L and 3.46 g/L in 3-L and 50-L fermenters, respectively, which were 69.54% and 75.63% higher than the yields under a fixed aeration strategy. This provides important technical support and an optimized plan for large-scale industrial production.

 

6.3 Economic feasibility study of large-scale G. lucidum production

G. lucidum is widely used in traditional Asian medicine for treating conditions such as hypertension, hepatitis, and cancer (Abdullah et al., 2020; Xiong et al., 2023). Its demand continues to grow in the global health market. However, traditional solid-phase cultivation of G. lucidum takes up to six months, is economically costly, and is prone to contamination, making large-scale commercial production unfeasible.

 

Araque et al. (2020) conducted a large-scale simulation of liquid fermentation for G. lucidum exopolysaccharides (EPS) using SuperPro Designer software to evaluate its economic feasibility. The results showed that the bioreactor volume significantly impacts production costs. In a 2-cubic-meter bioreactor, the cost per gram of product was $6.82, while in a 20-cubic-meter bioreactor, the cost dropped to $0.8 (Figure 3). Although genetically modified strains can increase EPS yields, scaling up production is the key to reducing costs. The study suggests that an optimal bioreactor volume of over 80 cubic meters is needed to achieve economically feasible production and meet market demand.

 

Figure 3 Q x Productivity vs EPS yield YEPS. (a) Volume 2 m3. (b) Volume 20 m3 y (c) Volume 20 m3 (Adopted from Araque et al., 2020)

Image caption: The figure shows the impact of different bioreactor volumes and exopolysaccharide (EPS) yields on the production efficiency of Ganoderma lucidum. The figure is divided into three parts, representing the relationship between production efficiency and EPS yield for 2-cubic-meter (a), 20-cubic-meter (b), and 200-cubic-meter (c) reactors. The results indicate that as the reactor volume increases, production costs decrease significantly, with the lowest cost observed in the 200-cubic-meter reactor. At the same time, the effect of EPS yield on costs is relatively small, suggesting that reactor volume is the primary factor in reducing production costs (Adapted from Araque et al., 2020)

 

7 Quality Control and Standardization

7.1 Quality assessment parameters

Ganoderma lucidum is renowned for its diverse array of bioactive compounds, which include polysaccharides, triterpenoids, and polyphenols. These compounds are primarily responsible for the mushroom's therapeutic properties, such as antioxidant, anti-inflammatory, anticancer, and hepatoprotective activities (Oke et al., 2022; Swallah et al., 2023). Polysaccharides, in particular, have garnered significant attention due to their high biotherapeutic properties, making them a critical quality indicator in Ganoderma lucidum products (Alsaheb et al., 2020; Kachrimanidou et al., 2023). Evaluating the purity and potency of Ganoderma lucidum products involves several analytical techniques. High-performance liquid chromatography (HPLC) and Fourier-transform infrared spectroscopy (FTIR) are commonly used to assess the chemical composition and confirm the presence of key bioactive compounds (Kachrimanidou et al., 2023). Additionally, the antioxidant activity of polysaccharides can be evaluated to determine their potency, ensuring that the products meet the desired therapeutic standards (Kachrimanidou et al., 2023).

 

7.2 Standardization of cultivation practices

Standardizing the cultivation practices of Ganoderma lucidum is essential to ensure consistent quality and yield of bioactive compounds. Guidelines for substrate preparation should include the use of agro-industrial by-products such as grape pomace and cheese whey, which have been shown to enhance biomass production and bioactive compound yield (Kachrimanidou et al., 2023). Environmental conditions, such as pH, temperature, and agitation, should be optimized to support the highest production of polysaccharides and other bioactive compounds (Alsaheb et al., 2020; Kachrimanidou et al., 2023). To ensure consistency across different production facilities, it is crucial to implement standardized protocols for substrate preparation, inoculation, and environmental control. This includes maintaining controlled pH conditions and optimizing carbon and nitrogen sources to achieve maximal yield (Alsaheb et al., 2020). Regular monitoring and quality checks should be conducted to ensure that all facilities adhere to these standardized practices, thereby guaranteeing uniformity in the final product quality.

 

7.3 Regulatory challenges in product quality

The regulatory landscape for medicinal mushroom products, including Ganoderma lucidum, varies significantly across different countries. International regulations often require rigorous safety assessments, quality assurance, and efficacy testing to ensure that the products meet the required standards for medicinal use (Swallah et al., 2023). Compliance with these regulations is essential for the global market acceptance of Ganoderma lucidum products. One of the major challenges in the standardization of Ganoderma lucidum products is the variability in global quality standards. Different countries may have varying requirements for the levels of bioactive compounds, purity, and potency. Addressing this variability requires a harmonized approach to quality control, which includes adopting standardized analytical methods and establishing international guidelines for the cultivation and processing of Ganoderma lucidum (Oke et al., 2022; Swallah et al., 2023). This will help in ensuring that the products are consistent and meet the therapeutic expectations across different markets.

 

8 Future Prospects and Research Directions

8.1 Innovations in cultivation technology

The future of Ganoderma lucidum cultivation lies in the continuous improvement and innovation of cultivation technologies. Recent studies have demonstrated the potential of submerged cultivation systems as an alternative to conventional solid-state fermentation, significantly enhancing the yield of bioactive compounds such as exopolysaccharides (EPS) (Alsaheb et al., 2020). Additionally, the development of bioreactors tailored for fungal liquid static culture has shown promise in optimizing the production of triterpenoids, particularly ganoderic acids, which are crucial for the medicinal value of G. lucidum (Hu et al., 2018). Strain improvement through methods such as ultraviolet ray irradiation has also been identified as a viable approach to increase spore yield and resistance to microbial invasions, further supporting industrial-scale production (Tang et al., 2023).

 

8.2 Addressing current production challenges

Despite the advancements in cultivation technology, several challenges remain in the industrial-scale production of G. lucidum. One significant issue is the impact of continuous cultivation on soil properties and microbial communities, which can affect the overall health and yield of the crops. Studies have shown that G. lucidum cultivation can significantly alter the microbial community structure and soil properties, necessitating the development of sustainable cultivation practices that mitigate these effects (Ren et al., 2020). Additionally, optimizing the nutritional profile and ensuring the consistency of bioactive compound production across different batches remain critical challenges. Comparative studies between different strains and cultivation conditions have highlighted the variability in antioxidant content, polyphenols, and protein digestibility, underscoring the need for standardized cultivation protocols (Fraile-Fabero et al., 2021).

 

8.3 Potential markets and applications

The diverse therapeutic and nutritional properties of G. lucidum open up numerous potential markets and applications. The mushroom's bioactive metabolites, such as polysaccharides, triterpenoids, and polyphenols, have been extensively studied for their antioxidant, anti-inflammatory, anticancer, and immunomodulatory activities, making G. lucidum a valuable resource for the pharmaceutical and nutraceutical industries (Swallah et al., 2023). The development of health products and pharmaceuticals derived from G. lucidum spore powder, which has shown significant pharmacological activities, represents a promising market opportunity (Tang et al., 2023). Furthermore, the mushroom's potential as a functional food ingredient, owing to its rich nutritional profile, positions it well within the food industry for the development of nutrient supplements aimed at preventing and managing chronic diseases (Swallah et al., 2023).

 

9 Concluding Remarks

The industrial-scale cultivation of Ganoderma lucidum presents several challenges and opportunities. Key findings include the optimization of bioprocesses for exopolysaccharides (EPS) production, which demonstrated that submerged cultivation systems could significantly enhance yield compared to traditional solid-state fermentation methods. Additionally, strain improvement through ultraviolet ray irradiation has been shown to increase spore powder yield and resistance to microbial invasions, providing a practical method for enhancing production efficiency. The impact of G. lucidum cultivation on soil and microbial community structures has also been highlighted, indicating significant changes in microbial diversity and soil properties. Comparative studies between different strains and cultivation conditions have revealed substantial variations in antioxidant content, polyphenols, and nutritional profiles, which are crucial for industrial applications. The development of optimized culture conditions and bioreactor designs for triterpenoid production has also been a significant advancement, demonstrating the potential for large-scale production.

 

The findings from these studies have several implications for the industrial-scale production of G. lucidum. The optimization of cultivation systems, such as the use of submerged fermentation and controlled pH conditions, can lead to higher yields of valuable bioactive compounds like EPS and triterpenoids, making the process more economically viable. Strain improvement techniques, such as UV irradiation, can enhance the yield and quality of spore powder, addressing the challenges of resource scarcity and variability in production. Understanding the impact of G. lucidum cultivation on soil and microbial communities can inform sustainable cultivation practices that maintain soil health and productivity. The variations in nutritional and bioactive compound profiles between different strains and cultivation conditions underscore the importance of selecting optimal strains and conditions for specific industrial applications. The use of agro-industrial by-products as feedstocks for G. lucidum cultivation also presents an opportunity for sustainable and cost-effective production, aligning with bioeconomy principles.

 

In conclusion, the industrial-scale cultivation of G. lucidum holds significant promise for the production of valuable bioactive compounds with numerous health benefits. The advancements in cultivation techniques, strain improvement, and bioprocess optimization provide a solid foundation for scaling up production while ensuring sustainability and economic feasibility. Future research should continue to explore innovative methods to enhance yield, quality, and sustainability, ensuring that G. lucidum remains a valuable resource for the pharmaceutical, nutraceutical, and food industries. The integration of these findings into industrial practices will be crucial for meeting the growing demand for G. lucidum products and maximizing their therapeutic potential.

 

Acknowledgments

We would like to sincerely thank Ms. Zhang from the Research Group of Ganoderma lucidum for her assistance in organizing the references. I am especially grateful to Dr. Li from the Traditional Chinese Medicine Research Group for revising the manuscript. We would also like to express my heartfelt thanks to the two anonymous peer reviewers for their thorough evaluations, which contributed to the improvement of the manuscript. 

 

Conflict of Interest Disclosure

The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.

 

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