Ganoderma lucidum fermentation liquor and preparation method thereof

Abstract

The invention belongs to the technical field of fungal fermentation, and particularly discloses ganoderma lucidum fermentation liquor and a preparation method thereof. The preparation method of the ganoderma lucidum fermentation liquor comprises the following steps: (1) activating a strain, namely inoculating the strain into a slant culture medium for culturing, and carrying out transfer grafting once to obtain an activated strain; (2) liquid culture: inoculating the activated strain into a liquid culture medium, and carrying out shake culture for 7 days to obtain a liquid strain; (3) fermentation culture: inoculating the liquid strain into a fermentation culture medium according to the inoculation amount of 7-9%, and carrying out three-stage culture to obtain a fermented product; and (4) centrifugal filtration: centrifuging the fermented product, collecting a supernatant, and filtering the obtained supernatant to obtain the ganoderma lucidum fermentation broth. According to the preparation method disclosed by the invention, three-stage variable-condition culture is specifically carried out according to the growth condition of the ganoderma lucidum mycelia, so that rapid growth of the ganoderma lucidum mycelia and mass accumulation of polysaccharides are facilitated, and the polysaccharide content of the prepared ganoderma lucidum fermentation liquor is increased.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of fungal fermentation, in particular to a Ganoderma lucidum fermentation liquor and a preparation method thereof. BACKGROUND

[0002] Ganoderma lucidum, belonging to Basidiomycota, Agaricomycetes, Polyporales and Ganodermataceae in taxonomy, is a rare edible and medicinal fungus, rich in various bioactive components. Ganoderma lucidum has a long history of medicinal use, and the research on its effective components and pharmacological effects has been extensive and in-depth, among which the research on Ganoderma lingzhi polysaccharide (GLP) is the most in-depth. Ganoderma lingzhi polysaccharide has the effects of immune regulation, anti-tumor, liver protection, blood lipid reduction and nerve cell protection, and is called "biological response regulator". Ganoderma lingzhi polysaccharide can be obtained from Ganoderma lucidum fruiting bodies, spore powder and Ganoderma lucidum fermentation products.

[0003] The artificial production of Ganoderma lucidum mainly includes solid culture method and liquid fermentation method. The solid culture method is convenient and economical, and can grow fruiting bodies, which is an effective method to obtain Ganoderma lucidum products; however, the solid culture method has the disadvantages of long cycle, difficulty in effectively determining the end of culture and strictly controlling the content and quality of the product. The liquid fermentation method is not affected by seasons, has a short culture period, low cost, can quickly proliferate fungal cells, and the production of effective components and active substances is easy to control, which has attracted widespread attention. Studies have shown that Ganoderma lingzhi polysaccharide produced by liquid fermentation has similar efficacy to polysaccharide derived from Ganoderma lucidum fruiting bodies and Ganoderma lucidum spores.

[0004] However, the yield of Ganoderma lingzhi polysaccharide obtained by liquid fermentation is not high, which limits the commercial development of Ganoderma lingzhi polysaccharide products. SUMMARY

[0005] In view of the above, it is necessary to provide a Ganoderma lucidum fermentation liquor and a preparation method thereof. The preparation method of the present application is three-stage variable condition culture according to the growth of Ganoderma lucidum mycelium, so as to facilitate the rapid growth of Ganoderma lucidum mycelium and the accumulation of polysaccharide, thereby improving the polysaccharide content of the prepared Ganoderma lucidum fermentation liquor.

[0006] To achieve the above purpose, the technical scheme adopted by the present application is:

[0007] A preparation method of a Ganoderma lucidum fermentation liquor, comprising the following steps:

[0008] (1) Strain activation: after the strain is inoculated in the slant medium and cultured, it is transferred and expanded once to obtain activated strain;

[0009] (2) liquid culture: the activated bacteria are inoculated in liquid culture medium and cultured for 7 days to obtain liquid bacteria;

[0010] (3) fermentation culture: the liquid bacteria are inoculated in fermentation culture medium at an inoculation amount of 7-9% to perform three-stage culture to obtain fermentation product, wherein the three-stage culture comprises:

[0011] first-stage culture: initial pH is 4.0-4.2, temperature is 25-26℃, stirring speed is 190 r / min, ventilation amount is 3.5 L / min, and culture time is 2 days;

[0012] second-stage culture: initial pH is 5.5-6.0, temperature is 30-31℃, stirring speed is 190 r / min, ventilation amount is 5 L / min, and culture time is 2 days;

[0013] third-stage culture: initial pH is 5.5-6.0, temperature is 26-27℃, stirring speed is 180 r / min, ventilation amount is 4 L / min, and culture time is 2 days;

[0014] (4) centrifugal filtration: the fermentation product is centrifuged to collect supernatant, and the obtained supernatant is filtered to obtain the ganoderma fermentation liquor.

[0015] Further, in step (1), the slant culture medium is PDA culture medium.

[0016] Further, in step (2), the liquid culture medium comprises 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O and 18 mg / g VB2.

[0017] Further, in step (3), the fermentation culture medium comprises 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate and 2 g / L yeast extract.

[0018] Further, the fermentation culture medium further comprises 3% malt extract.

[0019] The application further provides a ganoderma fermentation liquor prepared by the above preparation method.

[0020] The application has the following beneficial effects:

[0021] 1.The applicant of the present application found through a large amount of experimental data accumulation and analysis that targeted use of different growth conditions in different growth stages of Ganoderma lucidum is conducive to the production and accumulation of polysaccharides. Among them, the fermentation temperature is 25-26℃, the pH is 4.0-4.2, and the aeration rate is 3L / min in the first stage of mycelium growth. The relatively low temperature, pH and low aeration rate make the fermentation conditions relatively mild, which can effectively promote the cell activity and lay a solid foundation for subsequent growth. As the growth advances to the second stage, the mycelium proliferates rapidly, and the metabolic activity of the mycelium is enhanced. At this time, the fermentation temperature is increased to 30-31℃, and the pH and aeration rate are appropriately increased, which can provide the best conditions for the metabolic activity of the mycelium and the enzyme reaction of the cells, thereby promoting the growth and metabolism. In the late growth stage, the metabolic activity of the mycelium gradually slows down, and the fermentation temperature, pH and aeration rate are appropriately reduced. On the one hand, it can avoid the premature aging of the mycelium caused by the rapid enzyme inactivation, which makes it difficult to accumulate polysaccharides, and continue to accumulate polysaccharides to improve the content of polysaccharides. On the other hand, it can reduce the power consumption. At the same time, maintaining a high speed in the early and middle stages of mycelium growth can increase the proportion of small mycelial balls, thereby facilitating the synthesis of extracellular polysaccharides. In the late growth stage of the mycelium, the mycelial balls are weak, and the speed is reduced to avoid autolysis of the mycelium.

[0022] 2.In the fermentation medium of the present application, the addition of malt extract combined with yeast extract and peptone can provide more suitable chemicals and auxiliary factors for the growth and metabolism of Ganoderma lucidum mycelium, thereby further improving the content of polysaccharides in the Ganoderma lucidum fermentation broth. DETAILED DESCRIPTION

[0023] In order to make the technical problems, technical solutions and advantages of the present application more clear, the following will be described in detail in combination with specific examples.

[0024] Example 1

[0025] A preparation method of Ganoderma lucidum fermentation broth, comprising the following steps:

[0026] (1) Preparation of culture medium

[0027] Slope culture medium: PDA medium is selected;

[0028] Liquid culture medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, 18 mg / g VB2.

[0029] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, 2 g / L yeast extract.

[0030] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and expanded once, and cultured at 25°C for 7 days to obtain the activated strain; wherein the slant medium is the PDA medium.

[0031] (2) Liquid culture: the activated strain is inoculated in the liquid medium and placed in a rotary shaker for vibration culture at 26°C, 180 r / min for 7 days to obtain the liquid strain;

[0032] (3) Fermentation culture: the liquid strain is inoculated into the fermentation medium at a inoculation amount of 7% for three-stage culture to obtain the fermentation product, wherein the three-stage culture includes:

[0033] First-stage culture: initial pH 4.0, temperature 25°C, stirring speed 190 r / min, aeration amount 3.5 L / min, and culture time 2 days;

[0034] Second-stage culture: initial pH 5.5, temperature 30°C, stirring speed 190 r / min, aeration amount 5 L / min, and culture time 2 days;

[0035] Third-stage culture: initial pH 5.5, temperature 26°C, stirring speed 180 r / min, aeration amount 4 L / min, and culture time 2 days;

[0036] (4) Centrifugal filtration: the fermentation product is centrifuged at 3000 r / min for 20 min to separate, washed with the same volume of distilled water as the fermentation liquid, and centrifuged for 20 min to separate again, then the supernatants are combined, and the obtained supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

[0037] Example 2

[0038] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0039] (1) Preparation of culture medium

[0040] Slant medium: PDA medium is selected;

[0041] Liquid medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0042] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, 2 g / L yeast extract.

[0043] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and cultured for 7 days at 25°C to obtain the activated strain; wherein the slant medium is the PDA medium.

[0044] (2) Liquid culture: the activated strain is inoculated in the liquid medium and placed in a rotary shaker for vibration culture at 26°C and 180 r / min for 7 days to obtain the liquid strain.

[0045] (3) Fermentation culture: the liquid strain is inoculated into the fermentation medium at an inoculation amount of 8% for three-stage culture to obtain the fermentation product, wherein the three-stage culture comprises:

[0046] First-stage culture: initial pH 4.1, temperature 25.5°C, stirring speed 190 r / min, aeration amount 3.5 L / min, and culture time 2 days;

[0047] Second-stage culture: initial pH 5.8, temperature 30.5°C, stirring speed 190 r / min, aeration amount 5 L / min, and culture time 2 days;

[0048] Third-stage culture: initial pH 5.8, temperature 26.5°C, stirring speed 180 r / min, aeration amount 4 L / min, and culture time 2 days;

[0049] (4) Centrifugal filtration: the fermentation product is centrifuged at 3000 r / min for 20 min to separate, washed with the same volume of distilled water as the fermentation liquid, and centrifuged for 20 min to separate again, then the supernatants are combined, and the obtained supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

[0050] Example 3

[0051] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0052] (1) Medium preparation

[0053] Slant medium: PDA medium is selected;

[0054] Liquid medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0055] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, 2 g / L yeast extract.

[0056] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and cultured for 7 days at 25°C to obtain the activated strain; wherein the slant medium is the PDA medium.

[0057] (2) Liquid culture: the activated strain is inoculated in the liquid medium and placed in a rotary shaker for vibration culture at 26°C and 180 r / min for 7 days to obtain the liquid strain.

[0058] (3) Fermentation culture: the liquid strain is inoculated into the fermentation medium at an inoculation amount of 9% for three-stage culture to obtain the fermentation product, wherein the three-stage culture comprises:

[0059] First-stage culture: initial pH 4.2, temperature 26°C, stirring speed 190 r / min, aeration amount 3.5 L / min, and culture time 2 days;

[0060] Second-stage culture: initial pH 6.0, temperature 31°C, stirring speed 190 r / min, aeration amount 5 L / min, and culture time 2 days;

[0061] Third-stage culture: initial pH 6.0, temperature 27°C, stirring speed 180 r / min, aeration amount 4 L / min, and culture time 2 days;

[0062] (4) Centrifugal filtration: the fermentation product is centrifuged at 3000 r / min for 20 min to separate, washed with the same volume of distilled water as the fermentation liquid, and centrifuged for 20 min to separate again, then the supernatants are combined, and the obtained supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

[0063] Example 4

[0064] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0065] (1) Preparation of culture medium

[0066] Slant medium: PDA medium is selected;

[0067] Liquid medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0068] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, 2 g / L yeast extract, 3% malt extract.

[0069] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and cultured for 7 days at 25°C to obtain the activated strain; wherein the slant medium is the PDA medium.

[0070] (2) Liquid culture: the activated strain is inoculated in the liquid medium and placed in a rotary shaker for vibration culture at 26°C and 180 r / min for 7 days to obtain the liquid strain.

[0071] (3) Fermentation culture: the liquid strain is inoculated into the fermentation medium at an inoculation amount of 9% for three-stage culture to obtain the fermentation product, wherein the three-stage culture comprises:

[0072] First-stage culture: initial pH 4.2, temperature 26°C, stirring speed 190 r / min, aeration amount 3.5 L / min, and culture time 2 days;

[0073] Second-stage culture: initial pH 6.0, temperature 31°C, stirring speed 190 r / min, aeration amount 5 L / min, and culture time 2 days;

[0074] Third-stage culture: initial pH 6.0, temperature 27°C, stirring speed 180 r / min, aeration amount 4 L / min, and culture time 2 days;

[0075] (4) Centrifugal filtration: the fermentation product is centrifuged at 3000 r / min for 20 min to separate, washed with the same volume of distilled water as the fermentation liquid, and centrifuged for 20 min to separate again, then the supernatants are combined, and the obtained supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

[0076] Comparative Example 1

[0077] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0078] (1) Preparation of culture medium

[0079] Slant medium: PDA medium is selected;

[0080] Liquid medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0081] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, 2 g / L yeast extract.

[0082] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and cultured for 7 days at 25°C to obtain the activated strain; wherein the slant medium is the PDA medium;

[0083] (2) Liquid culture: the activated strain is inoculated in the liquid medium and cultured for 7 days at 26°C and 180 r / min in a rotary shaker to obtain the liquid strain;

[0084] (3) Fermentation culture: the liquid strain is inoculated in the fermentation medium at an inoculation amount of 9%, and cultured for 6 days under the conditions of initial pH 4.2, temperature 26°C, stirring speed 190 r / min, and ventilation amount 3.5 L / min;

[0085] (4) Centrifugal filtration: the fermentation product is centrifuged at 3000 r / min for 20 min to separate, washed with the same volume of distilled water as the fermentation liquid, and centrifuged for 20 min to combine the supernatant.

[0086] After centrifugal separation for 20 min, the obtained supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

[0087] Comparative Example 2

[0088] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0089] (1) Preparation of culture medium

[0090] Slant medium: PDA medium is selected;

[0091] Liquid medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0092] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, and 2 g / L yeast extract.

[0093] (2) Strain activation: the Ganoderma lucidum strain is inoculated in the PDA medium and cultured for 4 days, then transferred and cultured for 7 days at 25°C to obtain the activated strain; wherein the slant medium is the PDA medium;

[0094] (2) Liquid culture: inoculate the activated strain into liquid medium, and place it in a rotary shaker for oscillation culture at 26°C and 180 r / min for 7 days to obtain liquid strain;

[0095] (3) Fermentation culture: inoculate the liquid strain into fermentation medium at an inoculation amount of 9%, and culture for 6 days under the conditions of initial pH 6.0, temperature 28°C, stirring speed 180 r / min, and ventilation amount 5 L / min;

[0096] (4) Centrifugal filtration: centrifuge the fermentation product at 3000 r / min for 20 min to separate, wash with the same volume of distilled water as the fermentation liquid, continue to centrifuge for 20 min, and then combine the supernatant to obtain the Ganoderma lucidum fermentation liquid.

[0097]

[0098] Comparative Example 3

[0099] A preparation method of a Ganoderma lucidum fermentation liquid, comprising the following steps:

[0100] (1) Preparation of culture medium

[0101] Slope culture medium: PDA culture medium is selected;

[0102] Liquid culture medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

[0103] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, yeast extract 2 g / L, and 3% malt extract.

[0104] (2) Strain activation: inoculate the Ganoderma lucidum strain into PDA culture medium and culture for 4 days, then transfer and expand once, and culture at 25°C for 7 days to obtain activated strain; wherein the slope culture medium is PDA culture medium;

[0105] (2) Liquid culture: inoculate the activated strain into liquid medium, and place it in a rotary shaker for oscillation culture at 26°C and 180 r / min for 7 days to obtain liquid strain;

[0106] (3) Fermentation culture: inoculate the liquid strain into fermentation medium at an inoculation amount of 9% for two-stage culture to obtain fermentation product, wherein the two-stage culture comprises:

[0107] ​The first stage of culture: initial pH 4.2, temperature 26 °C, stirring speed 190 r / min, ventilation volume 3.5 L / min, culture time 3 days;

[0108] The second stage of culture: initial pH 6.0, temperature 31 ° C, stirring speed 190 r / min, ventilation volume 5 L / min, culture time 3 days;

[0109] (4) Centrifugal filtration: The fermented product was centrifuged at 3000 r / min for 20 min and the same volume of distilled water as the fermented liquid was used to separate the fermented product.

[0110] After washing, centrifugation was continued for 20 minutes, the supernatants were combined, and the obtained supernatants were filtered to obtain the Ganoderma lucidum fermentation liquid.

[0111] Comparative Example 4

[0112] A method for preparing a Ganoderma lucidum fermentation liquid comprises the following steps:

[0113] (1) Culture medium preparation

[0114] Slant culture medium: select PDA medium;

[0115] Liquid culture medium: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, 18 mg / g VB2.

[0116] Fermentation medium: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate.

[0117] (2) Activation of strains: The Ganoderma lucidum strains were inoculated into PDA medium and cultured for 4 days, then transferred once and cultured at 25°C for 7 days to obtain activated strains; wherein the slant culture medium is PDA medium;

[0118] (2) Liquid culture: The activated bacterial strain is inoculated into a liquid culture medium, and the culture is placed on a rotary shaker at 26°C and 180 rpm for 7 days to obtain a liquid bacterial strain;

[0119] (3) Fermentation culture: The liquid bacteria are inoculated into a fermentation medium at a rate of 9% and cultured in three stages to obtain a fermented product, wherein the three-stage culture comprises:

[0120] The first stage of culture: initial pH 4.2, temperature 26 °C, stirring speed 190 r / min, ventilation volume 3.5 L / min, culture time 2 days;

[0121] Second stage culture: initial pH 6.0, temperature 31℃, stirring speed 190r / min, aeration amount 5L / min, culture time 2d;

[0122] Third stage culture: initial pH 6.0, temperature 27℃, stirring speed 180r / min, aeration amount 4L / min, culture time 2d;

[0123] (4) Centrifugal filtration: centrifugal separation of the fermentation product at 3000r / min for 20min, washing with the same volume of distilled water as the fermentation liquid

[0124] , and centrifugal separation for another 20min, then combining the supernatant, filtering the obtained supernatant, and obtaining the Ganoderma lucidum fermentation liquid.

[0125] Effect verification

[0126] The Ganoderma lucidum fermentation liquids of the above examples 1-4 and comparative examples 1-4 were concentrated to 1 / 2 of the volume of the fermentation liquid, then precipitated with 5 times the volume of 95% ethanol, centrifuged for 20min, precipitated with 95% ethanol in the same volume as the concentrated liquid, centrifuged for another 20min, and dried at 60℃ to constant weight, which was the weight of the extracellular crude polysaccharide of each group, and the polysaccharide content (mg / 100mL) of each group of Ganoderma lucidum fermentation liquid was calculated, and the results are shown in Table 1.

[0127] Table 1 Polysaccharide content (mg / 100mL) of each group of Ganoderma lucidum fermentation liquid

[0128] Group Polysaccharide content (mg / 100 mL) Example 1 39.7 Example 2 40.6 Example 3 40.1 Example 4 42.8 Comparative Example 1 19.9 Comparative Example 2 24.7 Comparative Example 3 28.6 Comparative Example 4 31.2

[0129] As can be seen from Table 1, the polysaccharide content of examples 1-4 is significantly higher than that of comparative examples 1-4, indicating that the preparation method of the present application can effectively improve the polysaccharide content in the Ganoderma lucidum fermentation liquid, and the polysaccharide content of the fermentation liquid in example 4 is slightly higher than that in examples 1-3 because malt extract is added to the fermentation medium, which can provide more suitable chemical substances and auxiliary factors for the growth and metabolism of Ganoderma lucidum mycelium, thereby promoting the accumulation of polysaccharide. The polysaccharide content of comparative examples 1-3 is significantly reduced, indicating that the three-stage fermentation culture of the present application can indeed promote the growth and metabolism of Ganoderma lucidum, thereby improving the production and accumulation of polysaccharide. The polysaccharide content of comparative example 4 is also significantly reduced because the fermentation medium lacks yeast extract and malt extract, indicating that the fermentation medium combination of the present application can provide the most suitable conditions for the fermentation of Ganoderma lucidum.

Claims

1. A method for preparing Ganoderma lucidum fermentation liquid, characterized in that: The following steps are involved: (1) Activation of bacterial strains: After inoculating the bacterial strains into a slant culture medium and culturing them, they are transferred once to obtain activated bacterial strains; (2) Liquid culture: inoculating the activated bacterial strain into a liquid culture medium and culturing with shaking for 7 days to obtain a liquid bacterial strain; (3) Fermentation culture: The liquid bacteria are inoculated into a fermentation medium at an inoculum rate of 7-9% and cultured in three stages to obtain a fermentation product, wherein the three-stage culture comprises: The first stage of culture: initial pH 4.0-4.2, temperature 25-26 ° C, stirring speed 190 r / min, ventilation volume 3.5 L / min, culture time 2 days; The second stage of culture: initial pH 5.5-6.0, temperature 30-31 ° C, stirring speed 190 r / min, ventilation volume 5 L / min, culture time 2 days; The third stage of culture: initial pH 5.5-6.0, temperature 26-27 ° C, stirring speed 180 r / min, ventilation volume 4 L / min, culture time 2 days; (4) Centrifugal filtration: The fermented product is centrifuged to collect the supernatant, and the supernatant is filtered to obtain the Ganoderma lucidum fermentation liquid.

2. The method for preparing a Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: In step (1), the slant culture medium is PDA culture medium.

3. The method for preparing a Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: In step (2), the liquid culture medium includes: 5% glucose, 1% beef extract, 0.5% peptone, 0.2% ammonium sulfate, 0.1% KH2PO4, 0.1% MgSO4·7H2O, and 18 mg / g VB2.

4. The method for preparing a Ganoderma lucidum fermentation liquid according to claim 1, characterized in that: In step (3), the fermentation medium includes: 25% glucose, 0.7% peptone, 0.7% beef extract, 25 mg / g VB1, 0.05% KH2PO4, 0.09% MgSO4·7H2O, 0.15% ammonium sulfate, and 2 g / L yeast extract.

5. The method for preparing a Ganoderma lucidum fermentation liquid according to claim 4, characterized in that: The fermentation medium also includes 3% malt extract.

6. The Ganoderma lucidum fermentation liquid prepared by the preparation method according to any one of claims 1 to 5.