Phellinus linteus mycelium, and preparation method and application thereof

Abstract

The application belongs to the technical field of biological fermentation, and particularly discloses a phellinus igniarius mycelium, a preparation method and application thereof. The preparation method comprises the following steps: (1) activating the strain: inoculating phellinus igniarius mycelium pieces into PDA culture medium for culture to obtain activated phellinus igniarius; (2) seed culture: inoculating the activated phellinus igniarius into seed culture medium for culture to obtain phellinus igniarius seed liquid; (3) fermentation culture: transferring the phellinus igniarius seed liquid into fermentation culture medium for culture to obtain phellinus igniarius fermentation liquid; and (4) filtration and drying: filtering the phellinus igniarius fermentation liquid to obtain filter residue, washing and drying the filter residue to obtain the phellinus igniarius mycelium. The phellinus igniarius mycelium prepared by the method has high yield and high content of effective components, and can be used as raw material in health products, cosmetics or medicines.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation technology, and in particular to a Phellinus linteus mycelium, its preparation method, and its application. Background Technology

[0002] Sanghuangporus, belonging to the genus *Sanghuangporus*, is a type of perennial large fungus belonging to the phylum Basidiomycota, class Agaricales, order *Sanghuangporales*, and family *Sanghuangporaceae*. It is a valuable species of perennial large fungus used for both medicinal and edible purposes. Its fruiting bodies have a long history of medicinal use and are known as "forest gold," widely used in countries such as China, South Korea, and Japan. Related research indicates that the main chemical components of Sanghuang include polysaccharides, flavonoids, triterpenes, phenolic acids, and amino acids. These active substances exhibit good biological activity, lack cytotoxicity, and possess anti-cancer, immunomodulatory, hypoglycemic, antiviral, uric acid-lowering, antioxidant, and anti-inflammatory effects, making Sanghuang a potential area for new drug development.

[0003] However, wild Sanghuang (Phellinus linteus) resources are scarce, and artificial cultivation is difficult, resulting in relatively low yields. Furthermore, the extraction methods for fruiting bodies are contaminated, leading to high costs. Bio-fermentation technology, on the other hand, offers advantages such as a wide range of culture medium raw materials, low cost, short fermentation cycle, high yield, and good controllability, making it a more effective and safer method. During fermentation, the mycelium produces various secondary metabolites with anticancer, anti-inflammatory, and antibacterial activities, such as polysaccharides, flavonoids, polyphenols, and terpenoids, as well as abundant extracellular enzymes, showing promising research prospects and already widely used in the cultivation of the medicinal fungus Sanghuang. Therefore, exploring efficient and effective liquid fermentation processes for Sanghuang is of great significance for addressing the shortage of Sanghuang resources, developing its active ingredient products, and realizing its broad application prospects. Summary of the Invention

[0004] In view of the above, it is necessary to provide a method for preparing and applying Phellinus linteus mycelium. This invention uses a specific culture medium for a single fermentation treatment of Phellinus linteus, which can effectively increase the yield of Phellinus linteus mycelium and the content of its active ingredients, thus facilitating its subsequent application in pharmaceuticals.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] A method for preparing Phellinus linteus mycelium includes the following steps:

[0007] (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained.

[0008] (2) Seed culture: Activated Phellinus linteus was inoculated into seed culture medium and cultured at 26-27℃ and 200-400rpm for 4-5 days to obtain Phellinus linteus seed liquid;

[0009] (3) Fermentation culture: The seed liquid of Phellinus linteus is transferred to the fermentation medium at an inoculation rate of 8-10% (v / v) and cultured at 26-27℃ and 200-500rpm for 5-7 days to obtain Phellinus linteus fermentation broth;

[0010] The fermentation medium comprises the following components at the following mass concentrations: glucose 15-20 g / L, corn flour 5-8 g / L, yeast powder 5-8 g / L, wheat bran 1-2 g / L, magnesium sulfate 0.1-0.2 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L, mulberry extract 3-5 g / L, coix seed extract 2-5 g / L, and black tea extract 1-4 g / L;

[0011] (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, dried at 40-60℃ to constant weight, crushed and sieved to obtain Sanghuang mycelium.

[0012] Furthermore, in step (2), the seed culture medium comprises the following components at the following mass concentrations: glucose 15-20 g / L, yeast powder 3-5 g / L, peptone 3-5 g / L, magnesium sulfate 0.1-0.3 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L and mulberry extract 2-5 g / L.

[0013] Furthermore, in step (3), the mulberry extract is a water extract of mulberry, the coix seed extract is an ethanol extract of coix seed, and the black tea extract is an ethanol extract of black tea.

[0014] The present invention also provides a *Sanghuang* mycelium prepared by the above preparation method.

[0015] The present invention also provides a composition for the preparation of Phellinus linteus mycelium by the preparation method described above.

[0016] The present invention also provides the use of the *Sanghuang* mycelium or the composition in the preparation of pharmaceuticals.

[0017] The present invention has the following beneficial effects:

[0018] First, this invention utilizes liquid fermentation technology to solve the problem of scarcity of Sanghuang raw materials, and further avoids the instability of components caused by the depletion of wild resources and fluctuations in the cultivation environment. Second, in terms of process, this invention uses a specific culture medium to effectively increase the yield of Sanghuang mycelium, and at the same time, it can increase the content of effective components such as polysaccharides and flavonoids in Sanghuang mycelium, which is conducive to further development and utilization. Among them, mulberry trees, as one of the growth carriers of wild Phellinus linteus, contain various nutrients required for its growth. Therefore, its extracts can be used as components of the culture medium to promote Phellinus linteus growth. The extracts of coix seed and black tea also contain a variety of nutrients that can promote the growth of Phellinus linteus. At the same time, some nutrients in coix seed extract can bind to the cell membrane of Phellinus linteus, thereby enhancing its ability to absorb nutrients from the culture medium. More importantly, the mulberry tree extract, coix seed extract and black tea extract work together and complement each other, providing comprehensive and sufficient nutrition for the accumulation of various effective substances in Phellinus linteus, especially polysaccharides, flavonoids and triterpenes. This can promote the growth of Phellinus linteus mycelium and the accumulation of secondary metabolites, resulting in good activity. Detailed Implementation

[0019] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with specific embodiments.

[0020] In this invention, unless otherwise specified, all reagents and consumables used are purchased from conventional reagent manufacturers in the field; unless otherwise specified, all experimental methods and techniques used are conventional methods and techniques in the field.

[0021] The *Sanghuang* involved in this invention is a test-tube mother culture purchased from the Guangdong Lingnan Fungal Culture Factory on Taobao.

[0022] First, the present invention provides a Phellinus linteus mycelium and its preparation method.

[0023] In this embodiment, the method for preparing the *Phellinus linteus* mycelium includes the following steps:

[0024] (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained.

[0025] (2) Seed culture: Activated Phellinus linteus was inoculated into seed culture medium and cultured at 26-27℃ and 200-400rpm for 4-5 days to obtain Phellinus linteus seed liquid;

[0026] (3) Fermentation culture: The seed liquid of Phellinus linteus is transferred to the fermentation medium at an inoculation rate of 8-10% (v / v) and cultured at 26-27℃ and 200-500rpm for 5-7 days to obtain Phellinus linteus fermentation broth;

[0027] The fermentation medium comprises the following components at the following mass concentrations: glucose 15-20 g / L, corn flour 5-8 g / L, yeast powder 5-8 g / L, wheat bran 1-2 g / L, magnesium sulfate 0.1-0.2 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L, mulberry extract 3-5 g / L, coix seed extract 2-5 g / L, and black tea extract 1-4 g / L;

[0028] (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, dried at 40-60℃ to constant weight, crushed and sieved to obtain Sanghuang mycelium.

[0029] For example, in step (2), the culture temperature includes, but is not limited to: 26℃, 26.5℃, 26.8℃, 27℃, etc.; the rotation speed includes, but is not limited to: 200rpm, 230rpm, 250rpm, 280rpm, 300rpm, 330rpm, 360rpm, 380rpm, 400rpm.

[0030] Preferably, in step (2), the seed culture medium comprises the following components at the following mass concentrations: glucose 15-20 g / L, yeast powder 3-5 g / L, peptone 3-5 g / L, magnesium sulfate 0.1-0.3 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L and mulberry extract 2-5 g / L.

[0031] For example, in the seed culture medium:

[0032] The glucose concentration includes, but is not limited to: 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, and 20 g / L;

[0033] The mass concentration of the yeast powder includes, but is not limited to: 3 g / L, 3.2 g / L, 3.5 g / L, 4 g / L, 4.3 g / L, 4.5 g / L, and 5 g / L;

[0034] The peptone concentrations include, but are not limited to: 3 g / L, 3.3 g / L, 3.6 g / L, 4 g / L, 4.3 g / L, 4.7 g / L, and 5 g / L.

[0035] The mass concentration of the magnesium sulfate includes, but is not limited to: 0.1 g / L, 0.15 g / L, 0.2 g / L, 0.25 g / L, and 0.3 g / L;

[0036] The mass concentration of potassium dihydrogen phosphate includes, but is not limited to: 0.4 g / L, 0.45 g / L, 0.5 g / L, 0.55 g / L, and 0.6 g / L;

[0037] The mass concentration of the mulberry extract includes, but is not limited to: 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, 4 g / L, 4.5 g / L, and 5 g / L.

[0038] For example, in step (3):

[0039] The inoculation amount (v / v) of the Phellinus linteus seed liquid includes, but is not limited to: 8%, 8.5%, 9%, 9.5%, and 10%.

[0040] The fermentation temperatures include, but are not limited to, 26℃, 26.5℃, 26.8℃, and 27℃.

[0041] The fermentation rotation speeds include, but are not limited to: 200 rpm, 220 rpm, 250 rpm, 300 rpm, 350 rpm, and 400 rpm.

[0042] The fermentation time includes, but is not limited to: 5 days, 5.5 days, 6 days, 6.5 days, and 7 days.

[0043] For example, in the fermentation medium:

[0044] The glucose concentration includes, but is not limited to: 15 g / L, 16 g / L, 17 g / L, 18 g / L, 19 g / L, and 20 g / L;

[0045] The mass concentration of the corn flour includes, but is not limited to: 5 g / L, 6 g / L, 7 g / L, and 8 g / L;

[0046] The mass concentration of the yeast powder includes, but is not limited to: 5 g / L, 6 g / L, 7 g / L, and 8 g / L;

[0047] The mass concentration of the bran includes, but is not limited to: 1 g / L, 1.2 g / L, 1.5 g / L, 1.8 g / L, and 2 g / L;

[0048] The mass concentration of the magnesium sulfate includes, but is not limited to: 0.1 g / L, 0.13 g / L, 0.15 g / L, 0.18 g / L, and 0.2 g / L;

[0049] The mass concentration of potassium dihydrogen phosphate includes, but is not limited to: 0.4 g / L, 0.45 g / L, 0.5 g / L, 0.55 g / L, and 0.6 g / L;

[0050] The mass concentration of the mulberry extract includes, but is not limited to: 3 g / L, 3.5 g / L, 4 g / L, 4.5 g / L, and 5 g / L;

[0051] The mass concentration of the coix seed extract includes, but is not limited to: 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, 4 g / L, 4.5 g / L, and 5 g / L;

[0052] The mass concentration of the dark tea extract includes, but is not limited to: 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, and 4 g / L;

[0053] Preferably, the mulberry extract is a water extract of mulberry, the coix seed extract is an ethanol extract of coix seed, and the black tea extract is an ethanol extract of black tea.

[0054] For example, the preparation method of the mulberry water extract is as follows: the peeled mulberry branches are crushed and passed through an 80-mesh sieve, the mulberry powder and water are mixed at a solid-liquid ratio of 1:100g / L, extracted at 100℃ for 1h, cooled and filtered, and the obtained filtrate is concentrated under reduced pressure to about 15% of the original volume to obtain the mulberry extract, which is then stored at 4℃ for later use.

[0055] For example, the preparation method of the coix seed ethanol extract is as follows: coix seed is pulverized and passed through a 60-mesh sieve. The coix seed powder and 95% ethanol are mixed evenly at a material-to-liquid ratio of 1:4. The mixture is then extracted at 50°C for 30 minutes using a 300W ultrasonic wave. After cooling and filtration, the filtrate is concentrated under reduced pressure to about 50% of its original volume to obtain the coix seed extract. The extract is then stored at 4°C for later use.

[0056] For example, the preparation method of the black tea ethanol extract is as follows: add an 80% ethanol solution to black tea at a material-to-liquid ratio of 1:10 g / mL, heat and reflux for 2 hours, cool and filter, concentrate the obtained filtrate under reduced pressure to about 25% of the original volume, and the black tea extract is obtained. Store it at 4°C for later use.

[0057] For example, in step (4):

[0058] Drying temperatures include, but are not limited to: 40℃, 42℃, 45℃, 47℃, 50℃, 53℃, 55℃, 58℃, and 60℃.

[0059] The present application will be further described below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the present application.

[0060] Example 1

[0061] This embodiment provides a *Phellinus linteus* mycelium, the preparation method of which includes the following steps:

[0062] (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained.

[0063] (2) Seed culture: Activated Phellinus linteus was inoculated into seed culture medium and cultured at 26℃ and 200rpm for 5 days to obtain Phellinus linteus seed liquid;

[0064] The seed culture medium comprises the following components at the following mass concentrations: glucose 15 g / L, yeast powder 3 g / L, peptone 3 g / L, magnesium sulfate 0.1 g / L, potassium dihydrogen phosphate 0.4 g / L, and mulberry water extract 2 g / L.

[0065] (3) Fermentation culture: The seed liquid of Phellinus linteus was transferred to the fermentation medium at an inoculation rate of 8% (v / v) and cultured at 26℃ and 200rpm for 5 days to obtain Phellinus linteus fermentation broth;

[0066] The fermentation medium comprises the following components at the following mass concentrations: glucose 15 g / L, corn flour 5 g / L, yeast powder 5 g / L, wheat bran 1 g / L, magnesium sulfate 0.1 g / L, potassium dihydrogen phosphate 0.4 g / L, mulberry water extract 3 g / L, coix seed ethanol extract 2 g / L, and black tea ethanol extract 1 g / L.

[0067] (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, washed and centrifuged 3 times, dried at 40°C to constant weight, and crushed through a 50-mesh sieve to obtain the Sanghuang mycelium.

[0068] Example 2

[0069] This embodiment provides a *Phellinus linteus* mycelium, the preparation method of which includes the following steps:

[0070] (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained.

[0071] (2) Seed culture: Activated Phellinus linteus was inoculated into seed culture medium and cultured at 26.5℃ and 300rpm for 4.5 days to obtain Phellinus linteus seed liquid;

[0072] The seed culture medium comprises the following components at the following mass concentrations: glucose 18 g / L, yeast powder 4 g / L, peptone 4 g / L, magnesium sulfate 0.2 g / L, potassium dihydrogen phosphate 0.5 g / L, and mulberry water extract 4 g / L.

[0073] (3) Fermentation culture: The seed liquid of Phellinus linteus was transferred to the fermentation medium at an inoculation rate of 9% (v / v) and cultured at 27℃ and 400rpm for 6 days to obtain Phellinus linteus fermentation broth;

[0074] The fermentation medium comprises the following components at the following mass concentrations: glucose 18 g / L, corn flour 7 g / L, yeast powder 6 g / L, wheat bran 1.5 g / L, magnesium sulfate 0.15 g / L, potassium dihydrogen phosphate 0.5 g / L, mulberry water extract 4 g / L, coix seed ethanol extract 4 g / L, and black tea ethanol extract 3 g / L.

[0075] (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, washed and centrifuged 3 times, dried at 50°C to constant weight, and crushed through a 50-mesh sieve to obtain the Sanghuang mycelium.

[0076] Example 3

[0077] This embodiment provides a *Phellinus linteus* mycelium, the preparation method of which includes the following steps:

[0078] (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained.

[0079] (2) Seed culture: Activated Phellinus linteus was inoculated into seed culture medium and cultured at 27℃ and 400rpm for 4 days to obtain Phellinus linteus seed liquid;

[0080] The seed culture medium comprises the following components at the following mass concentrations: glucose 20 g / L, yeast powder 5 g / L, peptone 5 g / L, magnesium sulfate 0.3 g / L, potassium dihydrogen phosphate 0.6 g / L, and mulberry water extract 5 g / L.

[0081] (3) Fermentation culture: The seed liquid of Phellinus linteus was transferred to the fermentation medium at an inoculation rate of 10% (v / v) and cultured at 27℃ and 500rpm for 7 days to obtain Phellinus linteus fermentation broth;

[0082] The fermentation medium comprises the following components at the following mass concentrations: glucose 20 g / L, corn flour 8 g / L, yeast powder 8 g / L, wheat bran 2 g / L, magnesium sulfate 0.2 g / L, potassium dihydrogen phosphate 0.6 g / L, mulberry water extract 5 g / L, coix seed ethanol extract 5 g / L, and black tea ethanol extract 4 g / L.

[0083] (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, washed and centrifuged 3 times, dried at 60°C to constant weight, and pulverized through a 50-mesh sieve to obtain the Sanghuang mycelium.

[0084] Comparative Example 1

[0085] This comparative example provides a *Sanghuang* mycelium. The preparation method of the *Sanghuang* mycelium is different from that of Example 1, except that in step (2), the seed culture medium does not include mulberry water extract.

[0086] Comparative Example 2

[0087] This comparative example provides a *Sanghuang* mycelium. The preparation method of the *Sanghuang* mycelium differs from that of Example 1 only in that, in step (3), the fermentation culture medium does not include the ethanol extract of coix seed and the ethanol extract of black tea.

[0088] Comparative Example 3

[0089] This comparative example provides a *Sanghuang* mycelium. The preparation method of the *Sanghuang* mycelium differs from that of Example 1 only in that, in step (3), the fermentation culture medium does not include mulberry water extract and black tea ethanol extract.

[0090] Comparative Example 4

[0091] This comparative example provides a *Sanghuang* mycelium. The preparation method of the *Sanghuang* mycelium differs from that of Example 1 only in that, in step (3), the fermentation culture medium does not include mulberry water extract and coix seed ethanol extract.

[0092] Effect verification

[0093] 1. Determination of mycelial dry weight

[0094] Weigh the mycelium of each group of *Sanghuang* and calculate the dry weight of the mycelium. The results are shown in Table 1 below.

[0095] Table 1. Results of dry weight determination of Phellinus linteus mycelium in each group

[0096]

[0097] As shown in Table 1, the dry weight of Phellinus linteus mycelium in Examples 1-3 was significantly higher than that in Comparative Examples 1-4, indicating that the various components of the culture medium of the present invention work together to provide comprehensive nutrition for Phellinus linteus and effectively promote the growth of Phellinus linteus mycelium.

[0098] 2. Determination of polysaccharide yield

[0099] The polysaccharides were extracted from the mycelium of *Sanghuang* in Examples 1-3 and Comparative Examples 1-4 using the following method:

[0100] Anhydrous ethanol defatting → hot water extraction (extraction time 2 h, extraction temperature 100℃, material-liquid ratio 1:13) → filtration → 80% alcohol precipitation for 12 h → centrifugation → precipitate redissolution → dialysis → freeze drying → crude polysaccharide.

[0101] Calculate the polysaccharide yield of each group of Phellinus linteus mycelium using the following formula:

[0102] Polysaccharide yield (%) = (mass of crude polysaccharide after freeze-drying / dry mass of sample) × 100%;

[0103] The polysaccharide yields of various Phellinus linteus mycelia are shown in Table 2 below:

[0104] Table 2. Results of polysaccharide yield determination of Phellinus linteus mycelium in each group

[0105]

[0106] As shown in Table 2, when polysaccharides were extracted using the same extraction method, the polysaccharide yield of the *Sanghuang* mycelium in Examples 1-3 was significantly higher than that in Comparative Examples 1-4. This indicates that the addition of mulberry extract to the seed culture medium and the addition of a compound extract of mulberry, black tea, and coix seed to the fermentation culture medium can interact to jointly increase the content of active substances in *Sanghuang* mycelium.

[0107] 3. Antioxidant activity assay

[0108] The DPPH free radical scavenging capacity of crude polysaccharide (prepared at a concentration of 0.1 mg / mL) from *Sanghuang* mycelium in each group was determined. The results are shown in Table 3 below.

[0109] Table 3. Results of DPPH free radical scavenging capacity determination of polysaccharides from Phellinus linteus mycelium in each group.

[0110]

[0111] As shown in Table 3, the DPPH free radical scavenging rate of the polysaccharides from the mycelium of *Sanghuang* in Examples 1-3 was significantly higher than that in Comparative Examples 1-4. This indicates that the addition of mulberry extract to the seed culture medium and the addition of a compound extract of mulberry, black tea, and coix seed to the fermentation culture medium can interact to enhance the activity of polysaccharides in *Sanghuang* mycelium.

[0112] Based on the above effects, the present invention also provides a composition comprising the *Sanghuang* mycelium prepared by the above preparation method.

[0113] In summary, the Phellinus linteus mycelium or the composition containing Phellinus linteus mycelium of the present invention has excellent antioxidant properties. At the same time, it has been verified that Phellinus linteus mycelium also has anti-cancer, anti-inflammatory, antibacterial, and hypoglycemic biological activities, and can be used as a raw material in pharmaceuticals.

Claims

1. A method for preparing Phellinus linteus mycelium, characterized in that, Includes the following steps: (1) Activation of strain: The mycelial blocks of Phellinus linteus were inoculated into PDA medium for activation culture. After three generations of continuous activation culture, activated Phellinus linteus was obtained. (2) Seed culture: Activated Phellinus linteus is inoculated into seed culture medium and cultured at 26-27℃ and 200-400 rpm for 4-5 days to obtain Phellinus linteus seed liquid; the seed culture medium includes the following components at the following mass concentrations: glucose 15-20 g / L, yeast powder 3-5 g / L, peptone 3-5 g / L, magnesium sulfate 0.1-0.3 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L and mulberry extract 2-5 g / L; (3) Fermentation culture: The seed liquid of Phellinus linteus is transferred to the fermentation medium at an inoculation rate of 8-10% (v / v) and cultured at 26-27℃ and 200-500rpm for 5-7 days to obtain Phellinus linteus fermentation broth; The fermentation medium comprises the following components at the following mass concentrations: glucose 15-20 g / L, corn flour 5-8 g / L, yeast powder 5-8 g / L, wheat bran 1-2 g / L, magnesium sulfate 0.1-0.2 g / L, potassium dihydrogen phosphate 0.4-0.6 g / L, mulberry extract 3-5 g / L, coix seed extract 2-5 g / L, and dark tea extract 1-4 g / L; wherein the mulberry extract is an aqueous extract of mulberry, the coix seed extract is an ethanolic extract of coix seed, and the dark tea extract is an ethanolic extract of dark tea. (4) Filtration and drying: The fermentation liquid of Sanghuang is filtered to obtain filter residue. The filter residue is washed with distilled water, centrifuged, dried at 40-60℃ to constant weight, crushed and sieved to obtain Sanghuang mycelium.

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