TWI595089B - Industrial scale process of cultivating ganoderma lucidum mycelium - Google Patents

Description

Industrial grade culture method of ganoderma mycelium

The present disclosure relates to an industrial grade ganoderma lucidum culture method, particularly an industrial grade culture method for cultivating Ganoderma lucidum mycelium rather than Ganoderma lucidum fruiting bodies.

For a long time, in Southeast Asia, edible fungi have been used as nutritional supplements or health foods, among which Ganoderma Iucidum is a large one, and because of its medical effects, it has been used as a medicine for more than a thousand years. . The active ingredients isolated from Ganoderma lucidum generally include triterpenoids, polysaccharides, nucleic acids, polypeptides, and phytosterols. Among these active ingredients, Ganoderma lucidum triterpenoids are most important because of their pharmaceutically active activities, including inhibition of cholesterol synthesis, anticancer, antihypertensive and the like. Ganoderma lucidum triterpenoids include various compounds such as ganoderic acids (GAs), ganodermic acids (GMAs), ganoderic alcohols, ganoderic ketones, and ganoderic aldehydes. In a related study of ganoderic acid, it was found that ganoderic acid has the effect of killing cytotoxicity of cancer cells and inhibiting the amplification thereof. For example, ganoderic acid D (GAD) is known to prevent proliferation of human cervical cancer cells (ie, Hela cells) (Yue et al., Mol Cell Proteomics (2008) 7: 949-961); Acids A and H (GAA and GAH) can inhibit the growth of breast cancer cells or prevent them from invading other tissues (Jiang et al., Int J Mol Med (2008) 21: 577-584); Ganoderma acid X (ganoderic acid X, GAX) inhibits topographic enzyme activity and induces hepatoma cells to enter the apoptotic program (Li et al., Life Sci. (2005) 77, 252-265); Ganoderma lucidum M can effectively inhibit cancer In addition to cell growth, it also inhibits the migration of lung cancer cells (Wang et al., Int Immunopharmacol (2007) 7: 864-870), which is known to induce platelet aggregation (Wang et al., Biochim. Biophys. Acta. 1989) 986, 151-160), inhibition of platelet function (Wang et al., Biochem. J. (1991) 277 (Pt 1), 189-197) and inhibition by thromboxane A2 (Su et al., Biochem. Pharmacol. 1999) 58, 587-595; Su et al., Biochim. Biophys. Acta. (1999b) 1437, 223-234) or prostaglandin E1 (Su et al., Thromb. Res. (1999c) 99, 135-145) in platelets Induced cellular response

In the past, Ganoderma lucidum can only grow in the natural environment or only appear On the old trees in the remote mountains. However, as the cultivation technology of Ganoderma lucidum has been continuously improved, Ganoderma lucidum can now be cultivated in an artificial environment, but most of the solid substrate is used; for example, Japanese Laid-Open Patent No. 57014816 discloses a cultivation of Ganoderma lucidum by planting the spores of Ganoderma lucidum in wood. The method, but the disadvantage of this method is that the yield is too low, because it takes a lot of manpower to sow the spore itself, and the incubation period is too long, it takes 120 to 150 days to harvest, in addition, the grown Ganoderma lucidum and its contains The type and content of active compounds (triterpenoids) are far removed from natural ganoderma lucidum.

Based on this, there is an urgent need for an improved cultivation of Ganoderma lucidum. a method for producing a ganoderma for manufacturing a drug in a very short period of time Active compound.

Accordingly, it is an object of the present disclosure to provide an industrial grade cultivation method for cultivating Ganoderma lucidum, particularly for cultivating Ganoderma lucidum mycelium, rather than fruiting bodies. According to the present disclosure, up to about 15 kg of dried Ganoderma lucidum mycelium can be harvested from about 300 liters of liquid culture every 30 days, and the harvested dried Ganoderma lucidum mycelium is rich in high amounts of triterpenoids. It comprises at least ganoderic acid S (GAS), ganoderic acid T (GAT), ganoderic acid Me (GAMe), ganoderic acid R (GAR), and red spirit Acid S (ganodermic acid S, GMAS). According to a preferred embodiment, the Ganoderma lucidum mycelium cultivated by the method contains about 28-33 mg of a triterpenoid compound per gram of dry mycelium.

The disclosed industrial grade method for cultivating Ganoderma lucidum mycelium comprises the steps of: (a) inoculating the ganoderma mycelium in a liquid medium of no more than 1 liter and incubating for about 5-15 days; (b) using step (a) The harvested liquid culture is inoculated in at least 30 liters of liquid medium and incubated for about 2-5 days; (c) inoculation of the liquid culture harvested in step (b) in at least 300 liters of liquid medium and cultivating about 2 5 days; and (d) transferring the liquid culture collected in step (c) to a plurality of culture trays, each having a surface area of about 1.5 to 2.5 square centimeters per ml of liquid medium, and Incubate for about 10-20 days at about 200-2,000 illuminance.

According to a preferred embodiment, each step of the disclosed method The average is carried out at a temperature between about 20-35 ° C, a relative humidity of about 50-100%, and aeration of about 5-200 LPM (liters per minute).

According to a preferred embodiment, said steps (a), (b) and (c) Both were carried out in the dark with shaking or stirring at a speed of between about 50 and 200 rpm: and the illumination of step (d) was 720 lux.

Liquid medium suitable for use in the present invention comprises at least grapes Sugar, sucrose, peptone, yeast extracts and salts. Optionally, the liquid medium may further comprise soy flour.

According to a preferred embodiment, the method of the present invention further comprises The following steps: (e) harvesting the ganoderma mycelium from the liquid culture of the step (d); and (f) drying the ganoderma mycelium harvested in the step (e) until the moisture content thereof is less than 5%, wherein The dried Ganoderma lucidum mycelium contains about 28-33 mg of the triterpene compound per gram.

Dried Ganoderma lucidum obtained according to the cultivation method of the present invention The triterpene compound in the filament includes at least ganoderic acid S (GAS), ganoderic acid T (GAT), ganoderic acid Me (GAMe), and ganoderic acid R (ganoderic acid R, GAR), and ganodermic acid S (GMAS).

According to a preferred embodiment, the number of culture disks in step (d) The target is 288; and at least about 15 kg of dried Ganoderma lucidum mycelium can be harvested every 30 days in accordance with the method of the present invention.

Through the following detailed instructions, illustrations and accompanying application These and other features of the present disclosure will be more fully appreciated. The above summary and the following detailed description are merely illustrative, and are not intended to limit the scope of the disclosure.

100‧‧‧Cultivation system

110‧‧‧Shell

120, 130‧‧‧Cultivators

120a, 120b‧‧‧ column

121, 121a, 121b‧‧‧ plates

The above and other objects, features, advantages and embodiments of the present invention will become more apparent and understood. 2 is a schematic view of a culture tray 121a and a culture rack 120 according to another embodiment of the present invention.

The description of the embodiments of the present invention is intended to be illustrative and not restrictive. The features of various specific embodiments, as well as the method steps and sequences thereof, are constructed and manipulated in the embodiments. However, other specific embodiments may be utilized to achieve the same or equivalent function and sequence of steps.

Although numerical ranges and parameters are used to define a broad range of values for the present invention, the relevant values in the specific embodiments have been presented as precisely as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Here, "about" usually means the actual value is plus or minus 10%, 5% of a specific value or range, Within 1% or 0.5%. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the average, depending on the considerations of those of ordinary skill in the art to which the invention pertains. Except for the experimental examples, or unless otherwise explicitly stated, all ranges, quantities, values, and percentages used herein are understood (eg, to describe the amount of material used, the length of time, the temperature, the operating conditions, the quantity ratio, and the like. Are all modified by "about". Therefore, unless otherwise stated to the contrary, this specification and accompanying The numerical parameters disclosed in the scope of the patent application are approximate values and can be Change according to demand. At a minimum, these numerical parameters should be understood as the number of significant digits indicated and the values obtained by applying the general carry method.

The scientific and technical terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention pertains, unless otherwise defined herein. In addition, the singular noun used in this specification covers the plural of the noun in the case of no conflict with the context; the plural noun of the noun is also included in the plural noun used.

The present disclosure relates to a method for the industrial grade cultivation of Ganoderma lucidum mycelium, according to the disclosed method, at least about 15 kg of dried Ganoderma lucidum mycelium can be harvested every 30 days, wherein each g of dried Ganoderma lucidum mycelium contains up to about 28 -33 mg of a triterpene compound containing at least ganoderic acid S (GAS), ganoderic acid T (GAT), ganoderic acid Me (GAMe), and ganoderic acid R (ganoderic acid) R, GAR) and ganodermic acid S (GMAS). In addition, according to the present disclosure, if it is necessary to enlarge the yield, it is only necessary to scale up the disclosed batch process (ie, 2 times, 3 times or more) to obtain a proportional amount of dried Ganoderma lucidum mycelium. .

According to the disclosed method of cultivating the mycelium of Ganoderma lucidum, it comprises at least four stages. In the first stage, the Ganoderma lucidum mycelium is first inoculated into a culture flask containing not more than 1 liter of the first liquid medium, and cultured under appropriate culture conditions for about 5-15 days. Preferably, the ganoderma mycelium is inoculated to a first liquid medium of no more than 0.8 liters, more preferably no more than 0.5 liters. The disclosed first liquid medium comprises at least: 1.5% glucose, 1.5% sucrose, 0.5% peptone, 0.2% yeast extract, and 0.06% KH ₂ PO ₄ . After inoculation, the entire liquid medium is cultured under an environment of continuous shaking: an oscillation speed of about 50-120 rpm, for example about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115 or 120 rpm; temperature between about 20-35 ° C, for example about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 33, 34 or 35 ° C; for about 5-15 days, for example about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 days. Preferably, the oscillation speed is between about 65-105 rpm, such as about 65, 70, 75, 80, 85, 90, 95, 100 or 105 rpm; the temperature is between about 22-33 ° C, for example about 22, 23, 24 25, 26, 27, 28, 29, 30, 31, 32 or 33 ° C; for about 8-12 days, for example about 8, 9, 10, 11 or 12 days; more preferably, the oscillation speed is between about 75-90 rpm, such as about 75, 80, 85 or 90, rpm; temperature between about 25-30 ° C, such as about 25, 26, 27, 28, 29 or 30 ° C; incubation about 9-11 days, for example about 9 , 10 or 11 days. In a preferred embodiment, the incubation is carried out for about 10 days at an oscillation speed of about 85 rpm and a temperature of about 28 °C.

In the second stage, the liquid culture harvested in the first stage is first inoculated into a fermentation tank containing at least 30 liters of the first liquid medium. And incubated for about 2-5 days under appropriate culture conditions. Similarly, after inoculation, the entire liquid medium is cultured under constant agitation in an environment that satisfies the following conditions: agitation speed is between about 80 and 200 rpm, for example, about 80, 85, 90, 95, 100, 105, 110. , 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200 rpm; the amount of ventilation is between about 5-15 LPM, for example about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 LPM; and a temperature between about 20-35 ° C, such as about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 ° C; incubation for about 1-5 days, for example about 1, 2, 3, 4 or 5 days. Preferably, the agitation speed is between about 110 and 170 rpm, such as about 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165 or 170 rpm; the aeration is between about 7 and 12 LPM. , for example, about 7, 8, 9, 10, 11 or 12 LPM; temperature is between about 22 and 33 ° C, for example about 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or 33 ° C Incubating for about 2-4 days, for example about 2, 3 or 4 days; more preferably, the stirring speed is between about 120-160 rpm, for example about 120, 125, 130, 135, 140, 145, 150, 155 or 160 rpm. The aeration is between about 9-10 LPM, such as about 9 or 10 LPM; the temperature is between about 25-30 °C, such as about 25, 26, 27, 28, 29 or 30 °C; incubation is about 3 days. In a preferred embodiment, the incubation is carried out for about 3 days at a stirring speed of about 150 rpm, an aeration of about 10 LPM, and a temperature of about 28 °C.

In the third stage, the liquid culture harvested in the second stage is first inoculated into a fermenter containing at least 300 liters of the second liquid medium and incubated under appropriate culture conditions for about 2-6 days. The second liquid medium comprises at least: 3% glucose, 6.5% sucrose, 1.0% peptone, 3% soy flour, 0.6% yeast extract, 0.06% KH ₂ PO ₄ and 0.05% MgSO ₄ . Similarly, after inoculation, the entire liquid medium is cultured under constant agitation in an environment that satisfies the following conditions: agitation speed is between about 50-150 rpm, for example about 50, 55, 60, 65, 70, 75, 80. 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145 or 150 rpm; the amount of ventilation is between about 50-150 LPM, for example about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140 or 150 LPM; and a temperature between about 20-35 ° C, such as about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35 ° C; incubation for about 2-6 days, for example about 2, 3, 4, 5 or 6 days. Preferably, the agitation speed is between about 70-130 rpm, such as about 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125 or 130 rpm; the aeration is between about 80-130 LPM. , for example, about 80, 90, 100, 110, 120 or 130 LPM; temperature is between about 22 and 33 ° C, for example about 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or 33 ° C Incubating for about 3-5 days, for example about 3, 4 or 5 days; more preferably, the stirring speed is between about 85-115 rpm, for example about 85, 90, 95, 100, 105, 110 or 115 rpm; At about 90-110 LPM, for example about 90, 100 or 110 LPM; temperature between about 25-30 ° C, such as about 25, 26, 27, 28, 29 or 30 ° C; incubation for about 4 days. In a preferred embodiment, the incubation is carried out for about 4 days at a stirring speed of about 110 rpm, an aeration of about 100 LPM, and a temperature of about 28 °C.

In the fourth stage, the liquid culture harvested in the third stage is transferred to a plurality of culture trays, so that each milliliter of liquid in each culture tray The medium has a surface area of about 1.5-2.5 square centimeters and is then incubated at about 200-2,000 illuminance for about 10-20 days. In short, the liquid culture harvested in the third stage of about 0.5-1 liters is poured into each culture tray, and the area of each culture tray is between about 1,000-2,500 square centimeters, preferably between It is about 1,500-2,000 square centimeters, and more preferably about 1,800 square centimeters. In this stage, up to about 288 plates are used to hold the liquid culture harvested in the third stage for subsequent cultivation. Next, these culture trays containing the liquid culture harvested in the third stage are stacked, placed on a culture tray, pushed into an environment with temperature, humidity, ventilation, and light control to continue cultivation. In particular, the incubation temperature is controlled between about 20-35 ° C, such as about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35. °C; relative humidity is controlled between about 50-100%, such as about 50, 60, 70, 80, 90 or 100%; the amount of ventilation is between about 5-20 LPM, for example about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 LPM; light intensity between about 100 and 1,000 illuminance, for example about 100, 130, 200, 260, 300, 390, 400, 460, 520 , 650, 780, 910 or 1000 illuminance; incubation for about 10-20 days, for example about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 days. Preferably, the culture is maintained at a temperature of between about 22 and 33 ° C, such as about 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or 33 ° C; the relative humidity is controlled at about 70- 100%, for example about 70, 80, 90 or 100%; ventilation between about 8-18 LPM, such as about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 LPM; Intensity between about 200 and 780 illuminance, such as about 200, 260, 300, 390, 400, 460, 520, 650 or 780 illuminance; incubation for about 12-18 days, such as about 12, 13, 14, 15, 16, 17 or 18 days. Better yet Maintaining the culture at a temperature of between about 25 and 30 ° C, for example about 25, 26, 27, 28, 29 or 30 ° C; the relative humidity is controlled between about 80 and 100%, for example about 80, 90 or 100%; Between about 10-16 LPM, such as about 10, 11, 12, 13, 14, 15 or 16 LPM; light intensity between about 300 and 520 illuminance, such as about 300, 390, 400, 460 or 520 illuminance; incubation about 14- 16 days, for example about 14, 15 or 16 days. In a preferred embodiment, the incubation is for about 14 days at an aeration of about 15 LPM, a relative humidity of 100%, an illumination intensity of about 460 illuminance, and a temperature of about 28 °C.

Typically, the fourth stage of incubation is performed in the incubation system 100 as shown in Figure 1. This incubation system 100 includes a housing 110 and at least 4 culture racks; however, only two culture racks (120, 130) are shown in FIG. Each of the culture racks (120) respectively has two columns (120a, 120b), and each column can be respectively placed in at least one culture tray (121a). The culture tray can be made of a metal or plastic material and can have any shape or size. In the embodiment as shown in Fig. 2, the culture tray (121a) is made of stainless steel and has a rectangular shape with a width of about 40 cm, a length of about 60 cm, and a height of about 5 cm. Up to 36 plates can be placed in each column of each culture rack, so up to 72 plates can be stacked on each frame; therefore, the disclosed incubation system 100 can hold up to 288 plates.

Next, the Ganoderma lucidum mycelium cultivated and harvested according to the method disclosed above is air-dried at a temperature not higher than 70 ° C, preferably at a temperature not higher than 60 ° C until the moisture content thereof is less than about 10 Below %, preferably less than about 5%, more preferably less than about 3%.

In general, according to the above cultivation method, it can be harvested from each batch of culture or 300 liters of the third stage liquid culture. The dried Ganoderma lucidum mycelium of 10-20 kg is preferably a dried Ganoderma lucidum mycelium which is about 13-17 kg.

Moreover, in the dried Ganoderma lucidum mycelium harvested according to the present invention, up to about 28-33 mg of the triterpenoid compound per gram of dried Ganoderma lucidum mycelium; preferably, per gram of dried Ganoderma lucidum mycelium Contains up to about 30 mg of triterpenoids. These triterpenoid compounds include at least GAS, GAT, GAMe, GAR, and GMAS.

In addition, when it is desired to obtain higher amounts of triterpenoids, those skilled in the art having the ordinary knowledge can readily amplify the disclosed processes and systems by multiplying the disclosed methods and systems. Drying Ganoderma lucidum mycelium at times (eg, 2 times, 3 times or higher).

The following examples are presented to illustrate certain aspects of the present disclosure and to assist those skilled in the art to understand and practice the present disclosure. However, the scope of the disclosure is not limited to these embodiments.

Example

Example 1 Incubation of Ganoderma lucidum mycelium on an industrial scale

1.1 Production of the first stage liquid culture

About 1 square centimeter of Ganoderma lucidum mycelium was inoculated into 400 ml of a first liquid medium containing 1.5% glucose, 1.5% sucrose, 0.5% peptone, 0.2% yeast extract, and 0.06% KH ₂ PO ₄ . Next, the culture was continuously stirred at a rate of about 85 rpm and incubated at a temperature of about 28 ° C for about 10 days, and the resulting culture was referred to as "first stage liquid culture".

1.2 Production of the second stage liquid culture

Approximately 30 liters of the first liquid medium was poured into a fermentation tank having a volume of about 50 liters, and then about 2.8 liters of the first stage liquid culture of Example 1.1 was inoculated into the first liquid medium in the fermentation tank. . The culture was continuously stirred at a speed of about 150 rpm and incubated for about 3 days at aeration of about 10 LPM at a temperature of about 28 ° C. The resulting culture was referred to as "second stage liquid culture".

1.3 Production of the third stage liquid culture

About 300 liters of the second liquid medium was poured into a fermentation tank having a volume of about 500 liters, and then about 50 liters of the second stage liquid culture of Example 1.2 was inoculated into the second liquid medium in the fermentation tank. . The second liquid medium comprises at least: 3% glucose, 6.5% sucrose, 1.0% peptone, 3% soy flour, 0.6% yeast extract, 0.06% KH ₂ PO ₄ and 0.05% MgSO ₄ . Next, the culture was continuously stirred at a rate of about 100 rpm, and incubated at about 100 LPM and a temperature of about 28 ° C for about 4 days, and the resulting culture was called "third stage liquid culture".

1.4 Amplification and cultivation of the third stage liquid culture

In a sterile condition, pour about 1 liter of the third stage liquid culture into 288 plates (40 cm x 60 cm x 5 cm), and then place the plate into the culture as shown in Figure 1. The system was incubated for about 14 days at 100% relative humidity, aeration of about 15 LPM, temperature of about 28 ° C, and illumination of about 460 illuminance. The resulting mycelium of Ganoderma lucidum was collected and air dried at a temperature of about 60 ° C until its water content was less than about 5%.

Example 2 Effect of each incubation condition on the amount of triterpenoids in the dried Ganoderma lucidum mycelium of Example 1.

In this embodiment, by changing the incubation strip of embodiment 1.4 The amount of the triterpenoid compound contained in the product and the product was used to find the optimum conditions for amplifying the third stage liquid culture of Example 1.3. The incubation conditions tested and varied include: illumination intensity from 0 to 720 illuminance, ventilation from 0 to 30 LPM, relative humidity from 50% to 100%, and temperature from about 23 °C to 33 °C. Thereafter, the mycelium of Ganoderma lucidum cultivated under each condition was collected, air-dried at a temperature of 60 ° C until the moisture content was less than 5%, and the triterpenoid compound contained therein (which contained at least GAS, GAT, GAMe, GAR) was measured. And the total amount of GMAS). The results are shown in Tables 1-5.

Based on the results in Table 1-5 above, it can be found that if the Ganoderma lucidum culture in the final amplification stage is cultivated in a light environment, the content of triterpenoids in the mycelium of Ganoderma lucidum can be increased by up to 3 times, and the light in Table 1 is compared. As a result of illumination of 260 illuminance, the content of triterpenoids per gram of dry weight of mycelium increased from 9.11 mg to 26.66 mg. As for the effect of other incubation conditions on the content of triterpenoids, it was found that the ventilation should be at least 15 LPM, the relative humidity should be at least 80%, and the temperature should be at least 28 °C.

As for the number of cells cultured in the system, the experiment also found that when the number of cultured disks was small, the content of triterpenoids in the mycelium of Ganoderma lucidum decreased slightly. It is speculated that this is due to a decrease in relative humidity (ie, 80% cannot be maintained) due to a decrease in the culture in the cultivation system.

It will be understood that the above-described embodiments and examples are merely illustrative, and that various modifications may be made to the agent by those skilled in the art. The description, examples, and materials set forth above are intended to be illustrative of the present invention and are illustrative of the invention. The present disclosure has been disclosed in the above embodiments, but it is not intended to limit the disclosure, and any person skilled in the art can make various changes and refinements without departing from the spirit and scope of the disclosure. The scope of protection of the disclosure is subject to the definition of the scope of the patent application.

100‧‧‧Cultivation system

110‧‧‧Shell

120, 130‧‧‧Cultivators

120a, 120b‧‧‧ column

121a‧‧‧culture plate

Claims (9)

The invention relates to a method for cultivating Ganoderma lucidum mycelium, comprising: (a) inoculating a mycelium of Ganoderma lucidum in a liquid medium of not more than 1 liter and incubating for 5-10 days; (b) liquidating step (a) The culture is inoculated in at least 30 liters of the liquid medium and shaken for 2-5 days; (c) the liquid culture of step (b) is inoculated in at least 300 liters of the liquid medium and shaken for 2-5 days; And (d) dispensing the liquid culture of step (c) into a plurality of culture trays and incubating for 10-20 days at 200 to 2,000 illuminance, wherein in each culture tray, each milliliter of medium is about Has a surface area of 1.5-2.5 cm ^ 2 ; wherein steps (a), (b) and (c) are carried out in the dark, and any of steps (a) to (d) are at a temperature of 20-30 ° C Inter-, relative humidity between 50-100%, ventilation between 5-200LPM. The method of claim 1, wherein each of the culture disks has a surface area of about 1.8 square centimeters per milliliter of culture medium. The method of claim 1, wherein the liquid medium comprises glucose, sucrose, peptone, yeast extract, and salts. The method of claim 3, wherein the liquid medium is more Contains soy flour. The method of claim 2, wherein the illumination of step (d) is 460 illuminance. The method of claim 2, wherein the number of the culture disks is 288. The method of claim 2, further comprising: (e) harvesting the mycelium from the culture of step (d); and (f) drying the mycelium harvested in step (e) until the water content thereof is lower than 5% of the dried ganoderma lucidum mycelium, the total amount of triterpenoids per gram of Ganoderma lucidum mycelium dry weight is as high as 28-33 mg. The method of claim 7, wherein the triterpene compound comprises at least ganoderic acid S (GAS), ganoderic acid T (GAT), ganoderic acid Me (GAMe), and ganoderma lucidum. Acid R (ganoderic acid R, GAR), and Ganodermic acid S (GMAS). The method of claim 7, wherein the method produces about 15 kg of dried Ganoderma lucidum mycelium.