Ganoderma lucidum-roxburgh rose bidirectional fermentation liquor as well as preparation method and application thereof

Abstract

The invention discloses a ganoderma lucidum-roxburgh rose bidirectional fermentation broth and a preparation method and application thereof, and belongs to the technical field of roxburgh rose fermentation broth preparation. Ganoderma lucidum is used as an original strain, roxburgh rose is used as a matrix component, and ganoderma lucidum-roxburgh rose bidirectional liquid fermentation is performed; the antioxidant activity of the ganoderma lucidum fermentation liquid and the antioxidant activity of the ganoderma lucidum-roxburgh rose fermentation liquid and the antioxidant activity of the mycelium extraction liquid and the antioxidant activity of the artificially cultivated sporocarp extraction liquid are compared, so that the influence of roxburgh rose on ganoderma lucidum liquid fermentation is explored, and a certain thought is provided for diversified development and utilization and technical innovation of related new products with ganoderma lucidum and roxburgh rose as raw materials.

Description

Technical Field

[0001] The invention relates to the technical field of preparation of roxburghii fermented liquid, and in particular to a ganoderma-roxburghii bidirectional fermented liquid and a preparation method and application thereof. Background Art

[0002] Ganoderma lucidum belongs to the Ganodermaceae family and the Ganoderma genus. It is also known as fairy grass, Ganoderma grass, and Ruicao. It is a famous medicinal and edible fungus. Modern pharmacological research shows that Ganoderma has good physiological functions in anti-aging, liver protection, anti-tumor, anti-oxidation, and immune regulation. Rosa roxburghii Tratt, scientifically known as Rosa roxburghii, is a plant of the Rosaceae family. It is rich in active ingredients such as polysaccharides, flavonoids, and superoxide dismutase, and has multiple effects such as lowering blood sugar, anti-tumor, lowering blood lipids, and anti-oxidation.

[0003] At present, Ganoderma lucidum can be obtained through methods such as wild collection, artificial cultivation and fermentation. Wild collection is difficult and not easy to obtain, so the general production mainly adopts two methods: artificial cultivation of fruiting bodies and liquid fermentation. The cultivation methods of artificially cultivated fruiting bodies mainly include cultivation in simulated wild environments, bag cultivation, basswood cultivation, etc. Artificial cultivation of fruiting bodies takes a long time and requires high costs, and may cause insect and ant damage during the process. Studies have shown that the production of Ganoderma lucidum mycelium using liquid fermentation methods has the advantages of good growth conditions, short time required, easy large-scale fermentation, and low cost. In addition, during the deep liquid fermentation process, the enrichment of specific products can be achieved by regulating the fermentation conditions.

[0004] Bidirectional liquid fermentation is to add Chinese medicinal materials to the fungal fermentation system, and the Chinese medicinal materials can provide the nutrients needed for fungal growth, promote fungal fermentation, and produce more active ingredients under the action of fungi, so that the fermentation has bidirectionality. In recent years, the bidirectional fermentation of Ganoderma lucidum and other Chinese medicinal materials has been further developed. He Wei and others used astragalus, angelica, yam and coix seed to carry out liquid fermentation with Ganoderma lucidum. The four Chinese medicinal materials, especially astragalus, have a certain promoting effect on the liquid fermentation of Ganoderma lucidum, which has promoted the application of Ganoderma lucidum fermentation products in the fields of cosmetics and health care products. However, the fermentation level of the existing bidirectional fermentation of Ganoderma lucidum and other Chinese medicinal materials is relatively weak, and the content of active ingredients such as polysaccharides and polyphenols produced after fermentation is relatively low. To this end, the present invention provides a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid and its preparation method and application. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a Ganoderma lucidum-Roxbirica bidirectional fermentation liquid and a preparation method and application thereof, which effectively solves the technical problems that the existing fermentation level of Ganoderma lucidum and other Chinese medicinal materials is weak, and the content of active ingredients such as polysaccharides and polyphenols produced after fermentation is small, and at the same time provides a Ganoderma lucidum-Roxbirica bidirectional fermentation liquid fermentation process with short time consumption and higher antioxidant activity of the product.

[0006] The present invention provides a method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, which is characterized by comprising the following steps:

[0007] Inoculating the Ganoderma lucidum strain into a seed culture medium, and culturing at 26-30° C. with shaking to obtain a Ganoderma lucidum liquid seed solution;

[0008] Adding roxburghii powder to a fermentation medium with an initial pH of 5.0 to 6.5, sterilizing, and then adding the ganoderma lucidum liquid seed solution, shaking and culturing, to obtain a ganoderma lucidum-roxburghii bidirectional fermentation liquid;

[0009] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0010] As a preferred embodiment, each liter of the seed culture medium contains 35 g glucose, 0.5 g MgSO4·7H2O, 1 g KH2PO4, 5 g yeast extract powder, 0.1 g vitamin B1, 5 g peptone, and the rest is water.

[0011] As a preferred embodiment, 5 pieces of Ganoderma lucidum strains of 0.5 cm×0.5 cm×0.5 cm are inoculated into every 100 mL of the seed culture medium.

[0012] As a preferred embodiment, the amount of the roxburghii powder added is 16-20 g / L based on the volume of the fermentation medium.

[0013] As a preferred embodiment, the inoculation amount of the Ganoderma lucidum is 2% to 6% based on the volume of the fermentation medium.

[0014] As a preferred embodiment, the shaking culture is specifically: shaking culture at 150-180 r / min and 26-30° C. for 6-8 days.

[0015] As a preferred embodiment, the sterilization is specifically: sterilization at 121° C., 101-103 kPa for 25 minutes.

[0016] The second object of the present invention is to provide a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid prepared by the above preparation method.

[0017] The third object of the present invention is to provide an application of the above-mentioned Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid in the preparation of products with antioxidant activity.

[0018] As a preferred implementation, after keeping the temperature for 5 to 20 hours, the temperature is raised to 650 to 680° C. at a rate of 8.0° C. / min and kept for 1 to 3 hours, and then cooled along with the furnace.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The invention provides a ganoderma lucidum-roxburgh bidirectional fermentation liquid and a preparation method and application thereof. With ganoderma lucidum as a starting strain and roxburgh as a culture medium, the advantages of the ganoderma lucidum-roxburgh bidirectional liquid fermentation process and the antioxidant activity of the fermentation product are explored. When the initial pH value of the fermentation medium is 5.5, the ganoderma lucidum inoculation amount is 6%, and the roxburgh addition amount is 16 g / L, the polysaccharide content and biomass of the fermentation system are high; compared with the ganoderma lucidum fermentation liquid, after roxburgh is added to form a ganoderma lucidum-roxburgh bidirectional fermentation system, the fermentation level is improved, and the content of active substances polysaccharides, flavonoids, polyphenols and triterpenoid compounds is significantly increased; the ganoderma lucidum fermentation product and the ganoderma lucidum-roxburgh bidirectional fermentation product have an effect on the scavenging rate of DPPH free radicals, hydroxyl free radicals, and ABTS + The free radical scavenging rate and reducing power of Ganoderma lucidum fermentation product increased with the increase of mass concentration. When the mass concentration was 3 mg / mL, the hydroxyl radical scavenging rate and ABTS + The scavenging rate, DPPH free radical scavenging rate and reducing power of the liquid fermentation product of Ganoderma lucidum and Rosa roxburghii were 45.16%, 73.34%, 74.55% and 0.43 respectively, while those of the bidirectional fermentation product of Ganoderma lucidum and Rosa roxburghii were 51.55%, 94.79%, 84.87% and 0.60 respectively. Therefore, the bidirectional liquid fermentation product of Ganoderma lucidum and Rosa roxburghii was significantly higher than that of Ganoderma lucidum fermentation product, indicating that the addition of Rosa roxburghii promoted the improvement of the antioxidant activity of the fermentation system; the DPPH free radical scavenging capacity, ABTS + The free radical scavenging ability and hydroxyl radical scavenging ability were significantly higher than those of the artificially cultivated fruiting body extract, indicating that compared with artificially cultivated fruiting bodies, liquid fermentation has the advantages of shorter time and higher antioxidant activity of the products.

[0021] In this study, Ganoderma lucidum was used as the fermentation fungus and Rosa roxburghii was used as the culture ingredient. The antioxidant capacity and content of some antioxidant substances in the bidirectional fermentation products increased significantly, and the fermentation level was improved. This shows that the Ganoderma lucidum-Rosa roxburghii bidirectional liquid fermentation process has certain advantages, and provides certain ideas for the diversified development and utilization of related new products with Ganoderma lucidum and Rosa roxburghii as raw materials and technological innovation. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a graph showing the effect of the initial pH value of the fermentation medium of the present invention on the polysaccharide content and biomass;

[0023] Figure 2 This is a graph showing the effect of the amount of roxburghii added on the polysaccharide content and biomass of the present invention;

[0024] Figure 3 This is a graph showing the effect of the inoculation amount of Ganoderma lucidum on the polysaccharide content and biomass of the present invention;

[0025] Figure 4 A comparison chart of the scavenging abilities of the Ganoderma lucidum-Roxbirica roxb bidirectional fermentation liquid prepared in Example 1 of the present invention, the Ganoderma lucidum fermentation liquid of Comparative Example 1, and the vitamin C of Comparative Example 2 on hydroxyl radicals;

[0026] Figure 5 The results of the bidirectional fermentation liquid of Ganoderma lucidum and Rosa roxburghii prepared in Example 1 of the present invention, the fermentation liquid of Ganoderma lucidum prepared in Comparative Example 1 and the vitamin C in Comparative Example 2 on ABTS + Comparison chart of free radical scavenging ability;

[0027] Figure 6 A comparison chart of the scavenging ability of the Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid prepared in Example 1 of the present invention, the Ganoderma lucidum fermentation liquid of Comparative Example 1, and the vitamin C of Comparative Example 2 on DPPH free radicals;

[0028] Figure 7 A comparison chart of the total reducing capacity of the Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid prepared in Example 1 of the present invention and the Ganoderma lucidum fermentation liquid of Comparative Example 1;

[0029] Figure 8 The figure is a comparison of the scavenging rates of the fruiting body extract and the mycelium extract on hydroxyl radicals of the present invention;

[0030] Fig. 9 The fruiting body extract and mycelium extract of the present invention are used to treat ABTS + Comparison chart of free radical scavenging rates;

[0031] Fig.10 The figure is a comparison chart of the scavenging rates of the fruiting body extract and the mycelium extract on DPPH free radicals of the present invention. DETAILED DESCRIPTION

[0032] In order to enable those skilled in the art to better understand the technical solution of the present invention and implement it, the present invention is further described below in conjunction with specific examples, but the examples are not intended to limit the present invention. The following test methods and detection methods, unless otherwise specified, are conventional methods; the reagents and raw materials, unless otherwise specified, are commercially available.

[0033] The existing bidirectional fermentation of Ganoderma lucidum and other Chinese medicinal materials mentioned in the background technology of the present invention has been further developed. He Wei and others used astragalus, angelica, yam and coix seed to carry out liquid fermentation with Ganoderma lucidum. The four Chinese medicinal materials, especially astragalus, have a certain promoting effect on the liquid fermentation of Ganoderma lucidum, which has promoted the application of Ganoderma lucidum fermentation products in the fields of cosmetics and health care products. However, the fermentation level of the existing bidirectional fermentation of Ganoderma lucidum and other Chinese medicinal materials is relatively weak, and the content of active ingredients such as polysaccharides and polyphenols produced after fermentation is relatively low. Based on the above technical problems, the present invention provides a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid and its preparation method and application, using Ganoderma lucidum as the starting strain and Roxbirica roxbirica as the culture matrix, to explore the advantages of the Ganoderma lucidum-Roxbirica bidirectional liquid fermentation process and the antioxidant activity of its fermentation products.

[0034] The technical solution of the present invention is analyzed and explained in detail below.

[0035] The present invention first provides a method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, which is characterized by comprising the following steps:

[0036] The ganoderma lucidum strains are inoculated into a seed culture medium, and 5 pieces of ganoderma lucidum strains of 0.5 cm×0.5 cm×0.5 cm are inoculated into every 100 mL of the seed culture medium, and the culture is shaken at 26-30° C. to obtain a ganoderma lucidum liquid seed solution; a roxburgh powder is added to a fermentation medium with an initial pH of 5.0-6.5, and the culture is sterilized at 121° C. and 101-103 kPa for 25 min, and then the ganoderma lucidum liquid seed solution is added, and the culture is shaken at 150-180 r / min and 26-30° C. for 6-8 days to obtain a ganoderma lucidum-roxburgh bidirectional fermentation liquid;

[0037] In order to provide suitable carbon and nitrogen sources, the seed culture medium contains 35 g glucose, 0.5 g MgSO4·7H2O, 1 g KH2PO4, 5 g yeast extract powder, 0.1 g vitamin B1, 5 g peptone, and the rest is water per liter;

[0038] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0039] Calculated by the volume of the fermentation medium, the added amount of the roxburghii powder is 16-20 g / L, and the inoculation amount of the ganoderma lucidum is 2%-6%.

[0040] The technical effects of the present invention are described in detail below through specific embodiments and comparative examples.

[0041] Example 1

[0042] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0043] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0044] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 6% by volume roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0045] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0046] Example 2

[0047] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0048] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 26° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0049] Add 2.0 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.0, sterilize at 121° C. and 100 kPa for 25 min, then add the 4% by volume roxburghii inoculum to the roxburghii liquid seed solution, shake culture at 180 r / min and 26° C. for 6 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0050] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0051] Example 3

[0052] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0053] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 30° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0054] Add 1.8 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 6.0, sterilize at 121° C. and 102 kPa for 25 min, then add the 2% by volume roxburghii liquid seed solution, shake culture at 160 r / min and 30° C. for 8 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0055] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0056] Example 4

[0057] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0058] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0059] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 6.5, sterilize at 121° C. and 100 kPa for 25 min, then add the 6% by volume roxburghii inoculum to the roxburghii liquid seed solution, shake culture at 180 r / min and 28° C. for 8 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0060] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0061] Example 5

[0062] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0063] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 26° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0064] Add 2.0 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 4% by volume roxburghii liquid seed solution, shake culture at 150 r / min and 30° C. for 6 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0065] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0066] Example 6

[0067] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0068] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 30° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0069] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 6.0, sterilize at 121° C. and 103 kPa for 25 min, then add the 4% by volume roxburghii inoculum to the roxburghii liquid seed solution, shake culture at 150 r / min and 26° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0070] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0071] In order to further illustrate the technical effect of the present invention, the present invention also sets a comparative example, which is as follows:

[0072] Comparative Example 1

[0073] Compared with Example 1, the difference is that no Rosa roxburghii is added for bidirectional fermentation.

[0074] A method for preparing a ganoderma lucidum fermentation liquid, characterized in that it comprises the following steps:

[0075] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0076] 100 mL of fermentation medium with an initial pH of 5.5 was sterilized at 121° C. and 103 kPa for 25 min, and then the Ganoderma lucidum liquid seed solution was added according to a Ganoderma lucidum inoculation amount of 6% by volume, and cultured at 150 r / min and 28° C. for 7 days to obtain Ganoderma lucidum fermentation liquid;

[0077] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0078] Comparative Example 2

[0079] Vitamin C solutions with mass concentrations of 0.5 mg / mL, 1.0 mg / mL, 1.5 mg / mL, 2.0 mg / mL, 2.5 mg / mL, 3.0 mg / mL, 3.5 mg / mL and 4.0 mg / mL were used.

[0080] Comparative Example 3

[0081] Compared with Example 1, the difference is that the initial pH of the fermentation medium is 4.5.

[0082] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0083] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0084] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 4.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 6% by volume roxburghii inoculum to the roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0085] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0086] Comparative Example 4

[0087] Compared with Example 1, the difference is that the amount of the added Rosa roxburghii powder is reduced from 1.6 g to 1.2 g.

[0088] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0089] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0090] Add 1.2 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 6% by volume roxburghii inoculum to the roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0091] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0092] Comparative Example 5

[0093] Compared with Example 1, the difference is that the added amount of the roxburghii powder is reduced from 1.6 g to 0.8 g.

[0094] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0095] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0096] Add 0.8 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 6% by volume roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0097] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0098] Comparative Example 6

[0099] Compared with Example 1, the difference is that the added amount of the roxburghii powder is reduced from 1.6 g to 0.4 g.

[0100] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0101] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0102] Add 0.4 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 6% by volume roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0103] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0104] Comparative Example 7

[0105] Compared with Example 1, the difference is that the Ganoderma lucidum liquid seed solution is added according to a Ganoderma lucidum inoculation amount of 8% by volume.

[0106] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0107] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0108] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the 8% by volume roxburghii liquid seed solution, shake culture at 150 r / min and 28° C. for 7 days, and obtain a roxburghii-roxburghii bidirectional fermentation liquid;

[0109] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0110] Comparative Example 8

[0111] Compared with Example 1, the difference is that the Ganoderma lucidum liquid seed solution is added according to a Ganoderma lucidum inoculation amount of 10% by volume.

[0112] A method for preparing a Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid, characterized in that it comprises the following steps:

[0113] Inoculate 5 pieces of 0.5 cm×0.5 cm×0.5 cm Ganoderma lucidum strains into 100 mL of seed culture medium, and culture with shaking at 28° C. to obtain Ganoderma lucidum liquid seed solution; each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the rest is water;

[0114] Add 1.6 g of roxburghii powder to 100 mL of fermentation medium with an initial pH of 5.5, sterilize at 121° C. and 103 kPa for 25 min, then add the lingzhi liquid seed solution according to a 10% by volume inoculum of lingzhi, and culture at 150 r / min and 28° C. for 7 days to obtain lingzhi-roxburghii bidirectional fermentation liquid;

[0115] Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

[0116] The preparation process and performance test results of the Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid of the present invention are as follows.

[0117] Materials and reagents: Rosa roxburghii, produced in Guizhou; Ganoderma lucidum strains and Ganoderma lucidum cultivars, purchased from Minyuan strains; peptone and yeast extract powder, purchased from Beijing Solebow Co., Ltd.; Folin phenol, purchased from Shanghai MacLean Technology Co., Ltd.; thiamine hydrochloride, MgSO4·7H2O, ferric chloride, ABTS, rutin, oleanolic acid, sodium nitrite, ascorbic acid, phenol, sodium sulfite, ferrous sulfate, glucose, potassium ferrocyanide, KH2PO4, DPPH, trichloroacetic acid, analytical grade, purchased from Shanghai Aladdin Co., Ltd.

[0118] Instruments and equipment: SW-CJ-1F clean bench, Suzhou Antai Co., Ltd.; MQD-S3HR shaking incubator, Shanghai Minquan Instrument Co., Ltd.; BXM-30R vertical pressure steam sterilizer, Shanghai Boxun Medical Co., Ltd.; Alpha1-4LSC B freeze dryer, CHRIST, Germany; UV-1800PC ultraviolet-visible spectrophotometer, Shanghai Aoxiang Instrument Co., Ltd.

[0119] Determination of active ingredients

[0120] Ganoderma lucidum has high antioxidant activity, and its main active substances are polysaccharides and triterpenes. Rosa roxburghii contains a variety of active ingredients such as polysaccharides, polyphenols, and flavonoids, which also have good antioxidant activity. In order to explore whether the content of active substances in the fermented product will increase after adding rosa roxburghii powder, the content of polysaccharides, flavonoids, triterpenes and polyphenols in Ganoderma lucidum fermentation liquid and Ganoderma lucidum-rosa roxburghii fermentation liquid under the same fermentation conditions was determined.

[0121] Determination of polysaccharide content: Phenol-sulfuric acid method was used, glucose was used as the standard, and the regression equation was Y=10.4833x-0.052, R 2 =0.99925.

[0122] Flavonoid content determination: Rutin was used as the standard, and the regression equation was Y = 2.90574x-0.001, R 2 =0.99975.

[0123] Determination of polyphenol content: The polyphenol content was determined by the Folin phenol colorimetric method, with pyrogallol as the standard, and the regression equation was Y = 0.01558x-0.065, R 2 =0.99962.

[0124] Determination of triterpenoid content: The triterpenoid content was determined by vanillin-glacial acetic acid colorimetric reaction, oleanolic acid was used as the standard, and the regression equation was Y=4.64286x-0.02386, R 2 =0.99968.

[0125] Experimental investigation

[0126] 1. Effects of initial pH, amount of roxburghii added, and amount of Ganoderma lucidum inoculation on the Ganoderma lucidum-roxburghii bidirectional fermentation system

[0127] Taking the polysaccharide content and biomass of the fermentation system as indicators, the initial pH values ​​were set at 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0, the sea buckthorn addition amounts were 4g / L, 8g / L, 12g / L, 16g / L, 20g / L, and 24g / L, and the Ganoderma inoculation amount was 2%, 4%, 6%, 8%, and 10% (V / V) to explore the effects of different initial pH values, sea buckthorn addition amounts, and Ganoderma inoculation amounts on the Ganoderma lucidum-sea buckthorn bidirectional fermentation system.

[0128] 2. Cultivation and extraction of Ganoderma lucidum fruiting bodies

[0129] Keep it in an environment without direct sunlight, with good ventilation, temperature between 25 and 30 degrees Celsius, and humidity between 85% and 95%, and cultivate it for 60 to 90 days. The mature fruiting body of Ganoderma lucidum has a cylindrical shape, a cap that is close to a semicircular shape, a diameter of about 12 cm, and a thickness of about 1 to 2 cm. The surface of the mature fruiting body is relatively hard and has a certain glossiness. The whole Ganoderma lucidum is reddish brown, and the surface has ring-shaped and radial lines.

[0130] The obtained fruiting bodies were dried, crushed, and passed through a 40-mesh sieve. The water-to-liquid ratio was 1:30, the temperature was 90°C, the extraction time was 4h, the extraction times were twice, the two extracts were combined, concentrated by rotary evaporation at 50°C, and freeze-dried at low temperature to determine their antioxidant activity.

[0131] 3. Determination of Antioxidant Activity

[0132] 3.1 Hydroxyl free radical scavenging ability assay

[0133] Take 2 mL of sample solution with different mass concentrations, add 2.0 mL of 0.009 mol / L FeSO4 solution, 2.0 mL of 0.0088 mol / L H2O2 solution, and 2.0 mL of 0.009 mol / L salicylic acid solution, place in a water bath, react at 37°C for 30 min, vitamin C (VC) is used as a positive control, and the absorbance is detected at 510 nm.

[0134] Calculate according to formula (1):

[0135]

[0136] Where:

[0137] C1——hydroxyl radical scavenging rate;

[0138] A0 – absorbance of sample replaced by deionized water;

[0139] A1——absorbance of sample solution;

[0140] A2 – absorbance of deionized water instead of H2O2 solution.

[0141] 3.2ABTS + Free radical scavenging ability assay

[0142] Mix 0.0026mol / L K2S2O8 and 0.0074mol / L A BTS+ solution in a ratio of 1:1 and shake well. Store in a dark environment for 12 to 16 hours. Leave the mixed solution overnight and mix it with anhydrous ethanol in a certain proportion so that the absorbance of the mixed solution reaches about 0.70 at 734nm to obtain ABTS. + Working solution, prepare freshly before use. + The working solution was mixed evenly with 0.5 mL of sample solution of different mass concentrations, and reacted in the dark for 6 min at room temperature. The VC group was used as a positive control, and the absorbance was detected at 734 nm.

[0143] Calculate according to formula (2):

[0144]

[0145] Where:

[0146] C2——ABTS + Free radical scavenging rate;

[0147] A0——ABTS + Absorbance after mixing with deionized water working solution;

[0148] A1——Sample solution and ABTS + Absorbance of the working solution after mixing;

[0149] A2 – absorbance of the sample solution after mixing with deionized water.

[0150] 3.3 DPPH free radical scavenging ability determination

[0151] 2.0 mL of 0.0002 mol / L DPPH solution was mixed with 2 mL of sample solutions of different mass concentrations, shaken well, and allowed to stand for 30 min in a light-proof environment. The VC group was used as a positive control, and the absorbance was detected at 510 nm.

[0152] Calculate according to formula (3):

[0153]

[0154] Where:

[0155] C3——DPPH free radical scavenging rate;

[0156] A0——absorbance of the mixture of anhydrous ethanol and DPPH solution;

[0157] A1——absorbance of the mixture of sample solution and DPPH solution;

[0158] A2 – absorbance after mixing sample solution and anhydrous ethanol.

[0159] 3.4 Reducing power determination

[0160] Take 1.0mL of sample solution with different mass concentrations, add 2.5mL 0.2mol / L PBS solution and 2.5mL potassium ferricyanide solution, place in a water bath, react at 50℃ for 20min, take out, add 2.5mL 10% (W / V) trichloroacetic acid solution, shake well, centrifuge, take 2.5mL of the reaction supernatant into a centrifuge tube, add 2.5mL H2O and 0.5mL ferric chloride solution, mix well and let stand for 10min, and detect the absorbance value at 734nm.

[0161] Calculate according to formula (4):

[0162] C4=A1-A0 (4)

[0163] Where:

[0164] C4 – total reducing power;

[0165] A0——Deionized water replaces the absorbance of potassium ferrocyanide solution;

[0166] A1——Absorbance of sample solution.

[0167] 4. Data Processing

[0168] The data were expressed as mean ± standard deviation, and Excel 2019 was used for data processing and analysis.

[0169] Results and Discussion

[0170] 1. Content of active ingredients

[0171] As can be seen from Table 1, compared with the Ganoderma lucidum fermentation liquid, after adding Rosa roxburghii to form the Ganoderma lucidum-Rosa roxburghii bidirectional fermentation system, the content of active substances such as polysaccharides and flavonoids increased significantly, indicating that Rosa roxburghii effectively promoted the production of polysaccharides, flavonoids and other active substances during the growth of Ganoderma lucidum mycelium during the fermentation process. On the one hand, after Rosa roxburghii was added to the fermentation system, certain substances in Rosa roxburghii promoted the production of some active substances such as polysaccharides and triterpenes during the growth of Ganoderma lucidum mycelium during the fermentation process; on the other hand, Ganoderma lucidum strains will produce a large amount of enzymes during the fermentation process, and the produced enzymes may have a certain promoting effect on the decomposition of macromolecular substances such as cellulose in Rosa roxburghii tissues and cell walls, which is conducive to the precipitation of flavonoids and other substances in Rosa roxburghii, so that Rosa roxburghii can be used efficiently; secondly, the transformation of certain microorganisms in the Ganoderma lucidum-Rosa roxburghii fermentation system will also transform other components into substances with antioxidant activity.

[0172] Table 1 Comparison of active substance content in fermentation broth

[0173]

[0174] Note: Different lowercase letters indicate significant differences (P<0.05).

[0175] 2. Factors affecting the bidirectional fermentation of Ganoderma lucidum and Rosa roxburghii

[0176] 2.1 Effect of initial pH on polysaccharide content and biomass

[0177] The initial pH value of the culture medium will affect the morphology of the mycelium and the changes in the content of active substances during fungal growth. Different target products have different initial pH values. When the initial pH value of the culture medium is 4.5, 5.0, 5.5, 6.0, and 6.5, the experimental results are shown in Figure 1 When the initial pH value of the culture medium is less than 5.5, the polysaccharide content and biomass of the fermentation system increase with the increase of pH value. When the initial pH value of the culture medium is greater than 5.5, the polysaccharide content and biomass of the fermentation system decrease with the increase of pH value. When the pH value is 5.5, the polysaccharide content and biomass of the fermentation system reach peak values, which are 27.481 g / L and 3.1448 g / L respectively, which meets the most suitable growth conditions for Ganoderma lucidum at a pH value of 5.0-6.5.

[0178] 2.2 Effect of Rosa roxburghii addition on polysaccharide content and biomass

[0179] Since roxburghii contains a large amount of flavonoids, SOD, polysaccharides, VC and other active ingredients, adding roxburghii powder appropriately can increase the content of active ingredients in the culture medium and play a physical shearing effect on the mycelium during the shaking fermentation process. However, adding too much roxburghii powder will affect the dissolved oxygen content of the fermentation system and thus hinder the growth of Ganoderma lucidum. When the roxburghii addition amount is 4.0, 8.0, 12.0, 16.0, 20.0, 24.0 g / L, the experimental results show that Figure 2 When the amount of seabuckthorn added was less than 16.0g / L, the polysaccharide content and biomass of the fermentation system increased with the increase of the amount of seabuckthorn added. When the amount of seabuckthorn added was greater than 16.0g / L, the polysaccharide content and biomass of the fermentation system decreased with the increase of the amount of seabuckthorn added. When the amount of seabuckthorn added was 16.0g / L, the polysaccharide content and biomass of the fermentation system reached peak values, which were 23.4373g / L and 2.4539g / L, respectively. Therefore, the appropriate amount of seabuckthorn added to the fermentation system was 16.0g / L.

[0180] 2.3 Effect of Ganoderma lucidum inoculation amount on polysaccharide content and biomass

[0181] When the inoculation amount of Ganoderma lucidum is too low, the growth rate of the bacteria is slow, so it takes a long time to reach the end point of fermentation; when the inoculation amount of Ganoderma lucidum is relatively large, the bacteria will grow too vigorously, resulting in insufficient nutrients in the culture medium, leading to a decrease in the content of active ingredients in the fermentation system. When the inoculation amount of Ganoderma lucidum is 2%, 4%, 6%, and 8% (V / V), the experimental results are shown in Figure 3 , with the gradual increase of inoculation amount, the biomass and polysaccharide content of the fermentation system showed a trend of first rising and then falling, which was related to the size and number of mycelial balls. The inoculation amount had little effect on the biomass of the fermentation system, but had a greater impact on the content of active substances. Studies have shown that when the inoculation amount is large, the polysaccharide yield of the fermentation system is higher; when the inoculation amount is small, the mycelial ball diameter is larger, and the ganoderic acid yield in the fermentation system is higher. When the inoculation amount is 4% (V / V), the biomass reaches a peak of 2.6965g / L, and when the inoculation amount is 6% (V / V), the polysaccharide content of the fermentation system reaches the maximum, which is 25.7037g / L. Taking the polysaccharide content as the main indicator, 6% (V / V) is selected as the appropriate Ganoderma lucidum inoculation amount for the fermentation system.

[0182] 3. Evaluation of Antioxidant Activity of Bidirectional Fermentation Products

[0183] 3.1 Hydroxyl free radical scavenging ability

[0184] Depend on Figure 4In the range of 0.5-3.0 mg / mL, as the sample mass concentration gradually increased, the hydroxyl radical scavenging rates of Ganoderma lucidum fermented liquid and Ganoderma lucidum-roxburghii fermented liquid gradually increased. When the mass concentration was 3.0 mg / mL, the scavenging rate of Ganoderma lucidum fermented liquid was 45.16%, and the IC50 value was 3.34 mg / mL. The scavenging rate of Ganoderma lucidum-roxburghii fermented liquid was 51.55%, and the IC50 value was 2.74 mg / mL. The scavenging rate of VC on hydroxyl radicals gradually increased in the range of 0.5-1.0 mg / mL, reaching 99.93% at 1.0 mg / mL, and then basically maintained at this level. It can be seen that Ganoderma lucidum fermented liquid and Ganoderma lucidum-roxburghii fermented liquid have a certain scavenging ability on hydroxyl radicals. The addition of roxburghii to the fermentation system increased the hydroxyl radical scavenging rate, but the scavenging ability was weaker than that of VC.

[0185] 3.2 Ability to scavenge ABTS+ free radicals

[0186] Depend on Figure 5 It can be seen that in the range of 0.5-3.0 mg / mL, the ABTS+ free radical scavenging rates of Ganoderma lucidum fermentation broth and Ganoderma lucidum-roxburghii fermentation broth gradually increase with the increase of mass concentration. When the sample mass concentration is 3.0 mg / mL, the scavenging rate of Ganoderma lucidum fermentation broth is 73.34%, the IC50 value is 1.84 mg / mL, and the scavenging rate of Ganoderma lucidum-roxburghii fermentation broth is 94.79%, and the IC50 value is 1.08 mg / mL. The scavenging rate of VC on ABTS+ free radicals reaches 100% at 0.5 mg / mL, and then maintains at this level. When the sample mass concentration is 3.0 mg / mL, the scavenging power of Ganoderma lucidum-roxburghii bidirectional fermentation broth on ABTS+ is equivalent to that of VC. It can be seen that Ganoderma lucidum fermentation broth and Ganoderma lucidum-roxburghii fermentation broth have a certain scavenging ability on ABTS+ free radicals, and the addition of roxburghii improves the scavenging rate of ABTS+ free radicals in the fermentation system.

[0187] 3.3 DPPH free radical scavenging ability

[0188] Depend on Figure 6 It can be seen that in the range of 0.5-3.0 mg / mL, with the increase of sample mass concentration, the DPPH free radical scavenging ability of Ganoderma lucidum fermentation liquid and Ganoderma lucidum-roxburghii fermentation liquid gradually increased. When the sample mass concentration was 3.0 mg / mL, the scavenging rate of Ganoderma lucidum fermentation liquid was 74.55%, the scavenging rate of Ganoderma lucidum-roxburghii fermentation liquid was 84.87%, and VC reached 96.09% at 0.5 mg / mL, and then basically maintained at this level. It can be seen that Ganoderma lucidum fermentation liquid and Ganoderma lucidum-roxburghii fermentation liquid have a certain scavenging ability for DPPH free radicals, and the addition of roxburghii has a certain degree of improvement in the DPPH free radical scavenging rate of the fermentation system. When the sample mass concentration reached 3.0 mg / mL, the scavenging rate of Ganoderma lucidum-roxburghii fermentation liquid for DPPH free radicals began to approach VC.

[0189] 3.4 Total reduction capacity

[0190] Depend on Figure 7 It can be seen that within the range of 0.5-4.0 mg / mL, as the mass concentration of the sample increases, the reducing capacity of the Ganoderma lucidum fermentation liquid and the Ganoderma lucidum-Rosa roxburghii fermentation liquid gradually increases. When the mass concentration is 3.0 mg / mL, the reducing power of the Ganoderma lucidum fermentation liquid is 0.43, and the reducing power of the Ganoderma lucidum-Rosa roxburghii fermentation liquid is 0.60. It can be seen that the Ganoderma lucidum fermentation liquid and the Ganoderma lucidum-Rosa roxburghii fermentation liquid have a certain reducing capacity, and the addition of Rosa roxburghii improves the reducing capacity of the fermentation system.

[0191] 3. Comparison of antioxidant activity between artificially cultivated fruiting bodies and fermented mycelium extracts

[0192] Depend on Figure 8 to Figure 10 It can be seen that in the range of 1.0-6.0 mg / mL, the DPPH free radical scavenging rate, hydroxyl free radical scavenging rate, and ABTS + The free radical scavenging rate increased with the increase of mass concentration.

[0193] Depend on Figure 8 It can be seen that when the sample mass concentration is 6.0 mg / mL, the hydroxyl radical scavenging rate of the fruiting body extract is 22.77%, and the hydroxyl radical scavenging rate of the mycelium extract is 60.43%.

[0194] Depend on Fig. 9 It can be seen that when the sample mass concentration is 2.0 mg / mL, the ABTS content of the fruiting body extract is + The removal rate was 48.69% and the ABTS of mycelium extract + The clearance rate reached 100%.

[0195] Depend on Fig.10 It can be seen that when the sample mass concentration is 2.0 mg / mL, the DPPH free radical scavenging rate of the fruiting body extract is 42.27%, and the DPPH free radical scavenging rate of the mycelium extract reaches 92.69%.

[0196] The results showed that the DPPH free radical scavenging ability and ABTS + The free radical scavenging ability and hydroxyl radical scavenging ability are significantly higher than that of the artificially cultivated fruiting body extract. The content of active ingredients in the fruiting body and mycelium of the same Ganoderma lucidum strain is quite different, so it has different antioxidant activities.

[0197] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing a Ganoderma lucidum-Roxb. roxb. bidirectional fermentation liquid, characterized in that: The following steps are involved: Inoculating the Ganoderma lucidum strain into a seed culture medium, and culturing at 26-30° C. with shaking to obtain a Ganoderma lucidum liquid seed solution; Adding roxburghii powder to a fermentation medium with an initial pH of 5.0 to 6.5, sterilizing, and then adding the ganoderma lucidum liquid seed solution, shaking and culturing, to obtain a ganoderma lucidum-roxburghii bidirectional fermentation liquid; Each liter of the fermentation medium contains 35 g of glucose, 5 g of peptone, 1.0 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, and the rest is water.

2. The preparation method according to claim 1, characterized in that: Each liter of the seed culture medium contains 35 g of glucose, 0.5 g of MgSO4·7H2O, 1 g of KH2PO4, 5 g of yeast extract powder, 0.1 g of vitamin B1, 5 g of peptone, and the remainder is water.

3. The preparation method according to claim 1, characterized in that: Five pieces of Ganoderma lucidum strains measuring 0.5 cm×0.5 cm×0.5 cm were inoculated into every 100 mL of the seed culture medium.

4. The preparation method according to claim 1, characterized in that: Calculated by the volume of the fermentation medium, the added amount of the roxburghii powder is 16-20 g / L.

5. The preparation method according to claim 1, characterized in that: The inoculation amount of the ganoderma lucidum is 2% to 6% based on the volume of the fermentation medium.

6. The preparation method according to claim 1, characterized in that: The shaking culture specifically includes shaking culture at 150-180 r / min and 26-30° C. for 6-8 days.

7. The preparation method according to claim 1, characterized in that: The sterilization is specifically as follows: sterilizing at 121° C., 100-103 kPa for 25 minutes.

8. A Ganoderma lucidum-Roxb. roxb. bidirectional fermentation liquid prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the Ganoderma lucidum-Roxbirica roxbirica bidirectional fermentation liquid according to claim 8 in preparing a product with antioxidant activity.

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