White ginseng fungus fermentation product with soothing effect, preparation method and application thereof

Abstract

The present application discloses a soothing effect of white ginseng fungus fermentation product, and its preparation method and application. The preparation method of the soothing effect of white ginseng fungus fermentation product comprises the following steps: (1) fermentation substrate A comprises 100 parts of water, 0.4-5 parts of carbohydrate carbon source and 0.5-5 parts of organic nitrogen source; white ginseng fungus is inoculated into fermentation substrate A, and fermentation product A is obtained by aerobic fermentation for 24-48 hours; (2) fermentation substrate B comprises 100 parts of water, 0.4-2 parts of carbohydrate carbon source and 0.5-2 parts of organic nitrogen source; fermentation product A is mixed with fermentation substrate B, and a second aerobic fermentation is carried out for 12-20 hours, and sterilization is performed. The present application mainly adopts a two-step fermentation method, and strictly controls the fermentation time of each step, which is more conducive to the production of active ingredients by white ginseng fungus, so that the final product has excellent anti-inflammatory, antioxidant, damaged cell repair and anti-aging effects. It also achieves energy saving and environmental protection, and improves safety of use.

Description

Technical Field

[0001] The present application belongs to the field of biological fermentation, and specifically relates to a white ginseng fungus fermentation product with soothing effect, and a preparation method and application thereof. Background Art

[0002] White ginseng fungus, also known as Schizophyllum commune Fr., belongs to the Basidiomycetes and is a large edible and medicinal fungus. White ginseng fungus polysaccharide is the main active ingredient in white ginseng fungus. It has multiple physiological activities such as antibacterial, anti-inflammatory, and anti-radiation. It also has good skin moisturizing function and can delay skin aging to a certain extent. Compared with traditional moisturizers such as glycerin and sodium hyaluronate, it exhibits good moisture absorption and moisturizing properties.

[0003] There are reports in the prior art of using organic solvent ethanol to extract active ingredients from the fruiting body of white ginseng fungus. Extracting active ingredients from white ginseng fungus with organic solvents not only causes environmental pollution and increases production costs, but also requires the removal of the extraction solvent ethanol after extraction, which is not conducive to industrialization, and also leads to organic solvent residues, which poses certain safety issues and is not conducive to being used as a raw material for skin topical preparations.

[0004] Therefore, there is an urgent need in the art to develop a method for efficiently extracting active ingredients with excellent cosmetic effects from white ginseng fungus, with a simple process and high safety in the use of the obtained extract. Summary of the invention

[0005] The technical problem to be solved by the present application is to overcome the defects of the prior art that organic solvents are used to extract active ingredients from white ginseng fungi, the organic solvents need to be removed after extraction, the production cost is increased, and organic solvent residues will be caused, reducing the safety of product use, and provide a white ginseng fungi fermentation product with soothing effect, and its preparation method and application. The present application mainly adopts a two-step fermentation method, and strictly controls the fermentation time of each step, which is more conducive to the production of active ingredients by white ginseng fungi, and promotes the final product to have excellent anti-inflammatory, antioxidant, damaged cell repair and anti-aging effects. And there is no need to add organic solvents during the preparation process, so as to achieve energy saving and environmental protection, and effectively avoid organic solvent residues in the product, thereby improving the safety of product use.

[0006] This application adopts the following technical solutions to solve the above technical problems:

[0007] The present application provides a method for preparing a white ginseng fermentation product with soothing effect, which comprises the following steps:

[0008] (1) The fermentation substrate A comprises 100 parts of water, 0.4-5 parts of a carbohydrate carbon source and 0.5-5 parts of an organic nitrogen source; Schizophyllum commune is inoculated into the fermentation substrate A, and a fermentation product A is obtained by aerobic fermentation for 24-48 hours;

[0009] (2) The fermentation substrate B includes 100 parts of water, 0.4-2 parts of carbohydrate carbon source and 0.5-2 parts of organic nitrogen source; the fermentation product A is mixed with the fermentation substrate B, and the mixture is subjected to secondary aerobic fermentation for 12-20 hours, and sterilized to obtain the white ginseng fungus fermentation liquid.

[0010] In step (1) of some embodiments, the carbohydrate carbon source may include monosaccharides and / or starch-containing materials.

[0011] Wherein, the monosaccharide may include glucose.

[0012] Wherein, the starch-containing material may include at least one of rice flour, sorghum flour, corn flour and tapioca starch.

[0013] According to the conventional art, the preparation method of the rice flour comprises the following steps: the rice is dried, crushed, and sieved, wherein the mesh number of the sieve used for sieving can be 50 to 150 meshes, preferably 80 to 120 meshes, for example 100 meshes.

[0014] According to the conventional art, the preparation method of sorghum flour comprises the following steps: sorghum seeds are shelled, crushed, and sieved. The mesh number of the sieve used for sieving can be 50 to 150 meshes, preferably 80 to 120 meshes.

[0015] According to the conventional art, the preparation method of corn flour comprises the following steps: drying corn seeds, crushing, and sieving. The mesh number of the sieve used for sieving can be 50 to 150 meshes, preferably 80 to 120 meshes, such as 100 meshes.

[0016] In step (1) of some embodiments, the organic nitrogen source may include at least one of potatoes, soybean flour, black soybean flour, soybean meal and soybean peptide.

[0017] The potatoes may be peeled and shredded according to conventional procedures in the art before use.

[0018] The method for preparing soybean powder may include the following steps: shelling, drying, crushing and sieving soybeans, wherein the mesh size of the sieve used for sieving may be 50 to 150 meshes, preferably 80 to 120 meshes.

[0019] Wherein, the mass percentage of crude protein in the black bean powder can be 30% to 42% of the black bean powder.

[0020] The method for preparing black bean powder may include the following steps: shelling, drying, crushing and sieving black beans. The mesh size of the sieve used for sieving may be 50 to 150 meshes, preferably 80 to 120 meshes. In step (1) of some embodiments, the water may include deionized water commonly used in the art.

[0021] In step (1) of some embodiments, the weight proportion of the sugar carbon source is preferably 0.5 to 2 parts, more preferably 1 to 1.7 parts.

[0022] In step (1) of some embodiments, the weight proportion of the organic nitrogen source is preferably 0.5 to 2 parts, more preferably 0.8 to 1.5 parts.

[0023] In step (1) of some embodiments, the fermentation substrate A may be sterilized according to conventional procedures in the art before use.

[0024] The conditions and methods for sterilizing the fermentation substrate A may be high temperature sterilization methods conventionally used in the art.

[0025] When the high temperature sterilization method is used to sterilize the fermentation substrate A, the sterilization temperature can be a conventional temperature for such operations in the art, preferably 90-130°C, more preferably 115-125°C, for example 121°C.

[0026] When the high temperature sterilization method is used to sterilize the fermentation substrate A, the sterilization time can be the conventional time for such operation in the art, preferably 10 to 40 minutes.

[0027] According to the common practice in the art, the sterilization operation of the fermentation substrate A may further include a cooling operation, which may generally be cooling to room temperature.

[0028] In step (1) of some embodiments, the white ginseng fungus includes Schizophyllum commune Wbsh-01 (Schizophyllum commune) deposited in the General Microbiological Center of China Microorganism Culture Collection Administration with a deposit number of CGMCC No. 40388 and / or Schizophyllum commune purchased from China Industrial Microorganism Culture Collection Administration Center with a deposit number of CICC 2592.

[0029] Among them, the Schizophyllum commune Wbsh-01 (Schizophyllum commune) is deposited in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Administration, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, 100101, the deposit date is March 9, 2023, and the deposit number is CGMCC No.40388.

[0030] In step (1) of some embodiments, based on the water in the fermentation substrate A, the number of the white ginseng bacteria inoculated per unit mass of the water may be 10 5 CFU / g~10 7 CFU / g.

[0031] In step (1) of some embodiments, the white ginseng fungus can be added in the form of a white ginseng fungus liquid according to the conventional practice in the art.

[0032] Wherein, the live bacteria concentration of the white ginseng fungus in the white ginseng fungus liquid can be 10 6 ~10 8 CFU / mL.

[0033] The method for preparing the white ginseng fungus liquid may include the following steps: inoculating the white ginseng fungus into an activation culture medium, and subjecting the culture medium to activation culture.

[0034] In the preparation method of the white ginseng fungus liquid, the activation culture medium may include a PDA liquid culture medium. The PDA liquid culture medium may include 6g of potato, 20g of glucose and 1000mL of distilled water. The preparation method of the PDA liquid culture medium comprises the following steps: 6g of potato, 20g of glucose and 1000mL of distilled water are mixed and sterilized to obtain a mixture.

[0035] In the method for preparing the white ginseng fungus liquid, the temperature of the activation culture can be 25-35°C, preferably 25-30°C, such as 28°C.

[0036] In the method for preparing the white ginseng fungus liquid, the activation culture time can be 36 to 120 hours, preferably 36 to 96 hours, such as 72 hours.

[0037] In the method for preparing the white ginseng fungus liquid, the activation culture can be carried out under oscillation conditions according to the conventional art, and the oscillation speed can be 150-250 rpm, preferably 170-200 rpm, such as 180 rpm.

[0038] In step (1) of some embodiments, the time of the aerobic fermentation culture is preferably 35 to 48 hours, for example 36 hours.

[0039] In step (1) of some embodiments, the temperature of the primary aerobic fermentation culture may be 25-35°C, preferably 26-30°C, for example 28°C.

[0040] In step (1) of some embodiments, the primary aerobic fermentation culture can be carried out under oscillation conditions according to the conventional art, and the oscillation speed can be 150-250 rpm, preferably 170-200 rpm, for example 180 rpm.

[0041] In step (2) of some embodiments, the carbohydrate carbon source may include monosaccharides and / or starch-containing materials.

[0042] Wherein, the monosaccharide may include glucose.

[0043] Wherein, the starch-containing material may include at least one of rice flour, sorghum flour, corn flour and tapioca starch.

[0044] According to the conventional art, the preparation method of the rice flour comprises the following steps: the rice is dried, crushed, and sieved, wherein the mesh number of the sieve used for sieving can be 50 to 150 meshes, preferably 80 to 120 meshes, for example 100 meshes.

[0045] According to the conventional art, the preparation method of sorghum flour comprises the following steps: sorghum seeds are shelled, crushed, and sieved. The mesh number of the sieve used for sieving can be 50 to 150 meshes, preferably 80 to 120 meshes.

[0046] According to the conventional art, the preparation method of corn flour may include the following steps: drying corn seeds, crushing, and sieving. The mesh number of the sieve used for sieving may be 50 to 150 meshes, preferably 80 to 120 meshes, such as 100 meshes.

[0047] In step (2) of some embodiments, the organic nitrogen source may include at least one of potatoes, soybean flour, black soybean flour, soybean meal and soybean peptide.

[0048] The potatoes may be peeled and shredded according to conventional procedures in the art before use.

[0049] The method for preparing soybean powder may include the following steps: shelling, drying, crushing and sieving soybeans, wherein the mesh size of the sieve used for sieving may be 50 to 150 meshes, preferably 80 to 120 meshes.

[0050] Wherein, the mass percentage of crude protein in the black bean powder can be 30% to 42% of the black bean powder.

[0051] The method for preparing the black bean powder may include the following steps: shelling the black beans, drying, crushing, and sieving. The mesh size of the sieve used for sieving may be 50 to 150 meshes, preferably 80 to 120 meshes.

[0052] In step (2) of some embodiments, the water may include deionized water commonly used in the art.

[0053] In step (2) of some embodiments, the weight proportion of the carbohydrate carbon source is preferably 0.5 to 1.5 parts, more preferably 0.5 to 0.7 parts, for example 0.67 parts.

[0054] In step (2) of some embodiments, the weight proportion of the organic nitrogen source is preferably 0.8 to 2 parts, more preferably 0.8 to 1.67 parts.

[0055] In step (2) of some embodiments, the fermentation substrate B may be sterilized according to conventional procedures in the art before use.

[0056] The conditions and methods for sterilizing the fermentation substrate B may be high temperature sterilization methods conventionally used in the art.

[0057] When the high temperature sterilization method is used to sterilize the fermentation substrate B, the sterilization temperature can be a conventional temperature for such operations in the art, preferably 90-130°C, more preferably 115-125°C, for example 121°C.

[0058] When the high temperature sterilization method is used to sterilize the fermentation substrate B, the sterilization time can be the conventional time for such operation in the art, preferably 10 to 40 minutes, such as 30 minutes.

[0059] According to the common practice in the art, the sterilization operation of the fermentation substrate B may further include a cooling operation, which may generally be cooling to room temperature.

[0060] In step (2) of some embodiments, the secondary aerobic fermentation culture time is preferably 12 to 16 hours, such as 15 hours.

[0061] In step (2) of some embodiments, the temperature of the secondary aerobic fermentation culture may be 25-35°C, preferably 26-30°C, for example 28°C.

[0062] In step (2) of some embodiments, the secondary fermentation culture can be carried out under shaking conditions according to the conventional art, and the shaking speed can be 150-250 rpm, preferably 160-200 rpm, for example 180 rpm.

[0063] In step (2) of some embodiments, the sterilization method may be a high-temperature sterilization method conventionally used in the art.

[0064] When the high temperature sterilization method is used for the sterilization, the sterilization temperature may be 110-125°C, preferably 115-121°C.

[0065] When the high temperature sterilization method is used for the sterilization, the sterilization time may be 15 to 35 minutes, preferably 20 to 35 minutes, for example 30 minutes.

[0066] In step (2) of some embodiments, the sterilization operation may further include centrifugation and collection of the supernatant and / or mixing with a preservative.

[0067] The centrifugal speed may be 3000-9000 rpm, preferably 4000-6000 rpm, for example 4800 rpm.

[0068] The centrifugation time may be 10 to 40 min, preferably 20 to 40 min, for example 30 min.

[0069] Wherein, the radius of the centrifuge may be 8 to 15 cm, for example, 12 cm.

[0070] Wherein, during the process of mixing with the preservative, the mixing temperature may be 60-80°C, preferably 70-80°C.

[0071] Wherein, in the process of mixing with the preservative, the preservative used may include p-hydroxyacetophenone and / or 1,2-hexanediol.

[0072] When the preservative includes p-hydroxyacetophenone, the mass percentage of p-hydroxyacetophenone in the material obtained after sterilization is 0.3% to 1%, preferably 0.5%.

[0073] When the preservative includes 1,2-hexanediol, the mass percentage of 1,2-hexanediol in the material obtained after sterilization is 0.3% to 1%, preferably 0.5%.

[0074] The present application also provides a white ginseng fungus fermentation product with soothing effect, which is prepared by the preparation method of the white ginseng fungus fermentation product with soothing effect as described above.

[0075] The present application also provides a use of the white ginseng fermentation product with soothing effect as described above directly as a product, as an additive or as a base in the preparation of a skin external preparation.

[0076] In some embodiments, the soothing white ginseng fermentation product can be used as at least one of the anti-inflammatory active ingredient, antioxidant active ingredient, skin repair active ingredient and anti-aging active ingredient in the skin topical preparation.

[0077] The present application also provides a skin external preparation, which includes the white ginseng fungus fermentation product with soothing effect as described above.

[0078] In some embodiments, the skin topical preparation may further include active ingredients commonly used in the art.

[0079] The active ingredient may include at least one of a moisturizing active ingredient, a whitening active ingredient, an anti-inflammatory active ingredient, an anti-allergic active ingredient and an antioxidant active ingredient.

[0080] In some embodiments, the topical skin preparation may include but is not limited to a facial mask, an essence or a toner according to conventional methods in the art.

[0081] In some embodiments, the mass percentage of the soothing effect of the fermented product of white ginseng fungus in the skin external preparation may be 5% to 99%, preferably 60% to 99%.

[0082] In some embodiments, the room temperature generally refers to 24-30°C.

[0083] On the basis of conforming to the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present application.

[0084] The reagents and raw materials used in this application are commercially available.

[0085] The positive progress of this application is that: this application mainly adopts a two-step fermentation method and strictly controls the fermentation time of each step, which is more conducive to the production of active ingredients such as extracellular polysaccharides by white ginseng fungi, so that the final product has excellent anti-inflammatory, antioxidant, damaged cell repair and anti-aging effects. In addition, no organic solvents need to be added during the preparation process, which achieves energy conservation and environmental protection, and can effectively avoid organic solvent residues in the product, thereby improving the safety of product use. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] The present application can be better understood by referring to the description given below in conjunction with the accompanying drawings. The accompanying drawings, together with the following detailed description, are included in this specification and form a part of this specification, and are used to further illustrate the preferred embodiments of the present application and explain the principles and advantages of the present application.

[0087] in:

[0088] Figure 1 Schematic diagram of the ITS sequence phylogenetic tree of strain Wbsh-01 in Example 1;

[0089] Figure 2 This is a comparison chart of the expression of the inflammatory factor IL-8 in cells after the products prepared in Examples 2 to 7 and Comparative Examples 1 to 5 were used to treat cells stimulated with LPS;

[0090] Figure 3 A comparison chart of the DPPH free radical scavenging abilities of the products obtained in Examples 2 to 7 and Comparative Examples 1 to 5;

[0091] Figure 4 A comparison chart of the hydroxyl radical scavenging abilities of the products obtained in Examples 2 to 7 and Comparative Examples 1 to 5;

[0092] Figure 5 This is a comparison chart of cell viability after the products prepared in Examples 2 to 7 and Comparative Examples 1 to 5 treated cells damaged by ultraviolet light;

[0093] Figure 6 This is a comparison chart of the cell viability of human skin fibroblasts after being treated with the products prepared in Examples 2 to 7 and Comparative Examples 1 to 5. DETAILED DESCRIPTION

[0094] The present application is further described below by way of examples, but the present application is not limited to the scope of the examples. The experimental methods without specific conditions in the following examples are selected according to conventional methods and conditions, or according to the product instructions.

[0095] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0096] In the following examples, the PDA liquid culture medium raw material was purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd., the product number is HB0233-4, and the product name is potato glucose water.

[0097] The preparation method of PDA liquid culture medium is as follows: take 26g of potato glucose water (including 6g of potato powder and 20g of glucose), add it into 1000mL of deionized water, mix well, sterilize it in a high-temperature sterilizer at 121°C for 20min, and set aside.

[0098] Example 1

[0099] 1. Isolation of Schizophyllum Wbsh-01

[0100] Wild fungal fruiting bodies were collected from dead wood in Yunnan Province, and tissues at the junction of the cap and the stipe were picked, then placed on a slant culture medium (PDA plus enriched culture medium), and placed in a 28°C constant temperature incubator for dark culture for 5 days to obtain mycelium. The mycelium was picked up and purified three times using an inoculation loop to obtain a purified strain; the purification step was to pick the mycelium onto a slant culture medium (PDA plus enriched culture medium), and place it in a 28°C constant temperature incubator for dark culture for 5 days.

[0101] 2. Morphological observation of Schizophyllum Wbsh-01

[0102] Mycelium hyphae are white, branched, curved, and 1-2 μm in diameter. The hyphae grow radially around the inoculation point, and the hyphae at the inoculation point are yellow. When the mycelium develops to 8 days, the hyphae are entangled with each other, forming white protrusions on the surface of the substrate, which are the primordia of the fruiting body.

[0103] 3. Molecular biological identification of Schizophyllum Wbsh-01 (sent to Beijing New Era Zhonghe Technology Co., Ltd. for sequencing)

[0104] According to the operating procedures of the fungal genomic DNA extraction kit (model D3390-02, manufacturer: OMEGA), the genomic DNA of Schizophyllum sp. Wbsh-01 was extracted.

[0105] The genomic DNA of Schizophyllum Wbsh-01 was used as the amplification template, and the universal primers ITS1 (TCCGTAGGTGAACCTGCGG) and ITS4 (TCCTCCGCTTATTGAT ATGC) of the fungal ribosomal rDNA region were used as primers for PCR reaction (the manufacturer of the PCR instrument was Hangzhou Langji Scientific Instrument Co., Ltd., instrument model MG96+), and the PCR amplification program was as follows: 94°C pre-denaturation for 3 minutes to enter the cycle, 94°C pre-denaturation for 30 seconds, 54°C annealing for 30 seconds, 72°C extension for 90 seconds, for a total of 25 cycles. The obtained PCR product was sequenced, and the obtained rDNA-ITS sequence was directly compared with the publicly available rDNA-ITS sequence in the NCBI Genbank database for homology. The full-length ITS sequence was intercepted using TS2Database (as shown in Sequence Table 1), and the ITS phylogenetic tree analysis (see Figure 1 , Figure 1 Among them, 521 is Schizophyllum commune Wbsh-01), and the sequence similarity between strain Wbsh-01 and Schizophyllum commune MH539647.1 is 100%, indicating that the strain is Schizophyllum comm une.

[0106] Based on the results of morphological identification and DNA sequence analysis, strain Wbsh-01 was identified as Schizohyllum commune, belonging to the phylum Basidiomycota, the order Agaricales, and the genus Schizophyllum.

[0107] 4. Preservation of Schizophyllum commune strain Wbsh-01

[0108] The deposit date of the new strain Wbsh-01 is March 9, 2023, the deposit number is CGMCC No. 40388, the classification name is Schizophyllum commune, the name of the depository is the General Microbiology Center of China Culture Collection Administration (CGMCC for short), and the address is: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Postal Code 100101.

[0109] 5. Preparation of Schizophyllum Wbsh-01 bacterial solution

[0110] A single colony of Schizophyllum commune with a preservation number of CGMCC No. 40388 was picked up by an inoculation loop and inoculated into the above-configured PDA liquid culture medium, and activated and cultured for 72 hours at a temperature of 28° C. The activation culture was carried out under shaking conditions with an oscillation speed of 180 rpm to obtain a Schizophyllum commune bacterial solution; the live bacterial concentration of Schizophyllum commune in the Schizophyllum commune bacterial solution was 10 7 CFU / mL.

[0111] Example 2 Preparation of a white ginseng fermentation product having a soothing effect

[0112] (1) Shelling sorghum seeds, drying, grinding, and passing through a 100-mesh sieve to obtain sorghum flour, which is then set aside; peeling potatoes, cutting them into potato shreds, which is then set aside;

[0113] 3 g of sorghum flour, 3 g of potato shreds and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate A.

[0114] 30 mL of the bacterial solution of Schizophyllum Wbsh-01 prepared in Example 1 was inoculated into the fermentation substrate A prepared above. Based on the mass of water in the fermentation substrate A, the number of Schizophyllum Wbsh-01 inoculated per unit mass of water was 10 6 CFU / g; one aerobic fermentation culture was carried out at a temperature of 28°C for 48 hours, and one aerobic fermentation culture was carried out on an oscillator with a rotation speed of 180 rpm; after one aerobic fermentation culture, a fermentation product A was obtained;

[0115] (2) Shelling the black beans, drying, crushing, and passing through a 100-mesh sieve to obtain black bean powder for later use; the mass percentage of crude protein in the black bean powder is 30% to 42%;

[0116] 3 g of black bean powder, 1.5 g of glucose and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate B;

[0117] The fermentation product A prepared in step (1) and the fermentation substrate B prepared above are mixed, and secondary aerobic fermentation is carried out at a temperature of 28° C. for 15 hours, and the secondary aerobic fermentation is carried out on an oscillator with a rotation speed of 180 rpm; after the secondary aerobic fermentation, the mixture is sterilized at a temperature of 121° C. for 30 minutes, cooled to room temperature after sterilization, and centrifuged at a rotation speed of 4800 rpm and a centrifugal radius of 12 cm for 30 minutes, and the supernatant is taken to obtain a white ginseng fungus fermentation product with a soothing effect.

[0118] Example 3

[0119] Compared with Example 2, the only difference is that the time of the aerobic fermentation culture in step (1) is different, specifically 36 hours, and the other condition parameters are the same as Example 2, and a white ginseng fungus fermentation product with soothing effect is obtained.

[0120] Example 4

[0121] Compared with Example 2, the only difference is that the formula of the fermentation substrate A in step (1) is different. The fermentation substrate A includes 5g corn flour, 3g soybean peptide and 300g deionized water. The other condition parameters are the same as Example 2.

[0122] The preparation method of corn flour comprises the following steps: drying corn seeds, crushing them, and passing them through a 100-mesh sieve.

[0123] Example 5

[0124] Compared with Example 2, the only difference is that the formula of the fermentation substrate B in step (2) is different. The fermentation substrate B includes 2g rice flour, 5g soybean flour and 300g deionized water. The other condition parameters are the same as Example 2.

[0125] The preparation method of rice flour comprises the following steps: drying the rice, crushing it and passing it through a 100-mesh sieve.

[0126] The preparation method of soybean powder comprises the following steps: shelling soybeans, drying, crushing, and passing through a 100-mesh sieve.

[0127] Example 6

[0128] Compared with Example 2, the only difference is that the fermentation bacteria inoculated in step (1) are different, specifically Schizophyllum CICC 2592 is used, and other condition parameters are the same as those in Example 2;

[0129] In this example, Schizophyllum was purchased from China Industrial Microbiological Culture Collection Center with the collection number CICC2592.

[0130] The preparation method of the Schizophyllum commune bacterial liquid comprises the following steps: using an inoculation loop to pick a single colony of Schizophyllum commune with a preservation number of CICC 2592, and inoculating it into a PDA liquid culture medium, activating and culturing it at a temperature of 28° C. for 72 hours, wherein the activation culture is carried out under an oscillating condition with an oscillating speed of 180 rpm, to obtain the Schizophyllum commune bacterial liquid; the concentration of live bacteria of Schizophyllum commune in the Schizophyllum commune bacterial liquid is 10 7 CFU / mL.

[0131] Example 7

[0132] Compared with Example 2, the only difference is that the formula of the fermentation substrate A in step (1) and the fermentation substrate B in step (2) are different, and specifically includes the following steps:

[0133] (1) Shelling black beans, drying, crushing, and passing through a 100-mesh sieve to prepare black bean powder for later use; the mass percentage of crude protein in the black bean powder is 30% to 42%;

[0134] 3 g of black bean powder, 1.5 g of glucose and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate A.

[0135] 30 mL of the bacterial solution of Schizophyllum Wbsh-01 prepared in Example 1 was inoculated into the fermentation substrate A prepared above. Based on the mass of water in the fermentation substrate A, the number of Schizophyllum Wbsh-01 inoculated per unit mass of water was 10 6 CFU / g; one aerobic fermentation culture was carried out at a temperature of 28°C for 48 hours, and one aerobic fermentation culture was carried out on an oscillator with a rotation speed of 180 rpm; after one aerobic fermentation culture, a fermentation product A was obtained;

[0136] (2) Shelling sorghum seeds, drying, grinding, and passing through a 100-mesh sieve to obtain sorghum flour, which is then set aside; peeling potatoes, cutting them into potato shreds, which is then set aside;

[0137] 3 g of sorghum flour, 3 g of potato shreds and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate B;

[0138] The fermentation product A prepared in step (1) and the fermentation substrate B prepared above are mixed, and secondary aerobic fermentation is carried out at a temperature of 28° C. for 15 hours, and the secondary aerobic fermentation is carried out on an oscillator with a rotation speed of 180 rpm; after the secondary aerobic fermentation, the mixture is sterilized at a temperature of 121° C. for 30 minutes, cooled to room temperature after sterilization, and centrifuged at a rotation speed of 4800 rpm and a centrifugal radius of 12 cm for 30 minutes, and the supernatant is taken to obtain a white ginseng fungus fermentation product with a soothing effect.

[0139] Comparative Example 1

[0140] Compared with Example 2, the only difference is that the formula of the fermentation substrate A in step (1) is different. The fermentation substrate A includes 3g glycerol, 3g ammonium chloride and 300g deionized water. The other condition parameters are the same as Example 2.

[0141] Comparative Example 2

[0142] Compared with Example 2, the only difference is that the fermentation culture time in step (1) and step (2) is different. Specifically, the time of the first aerobic fermentation culture in step (1) is 15 hours, and the time of the second aerobic fermentation culture in step (2) is 48 hours. Other condition parameters are the same as in Example 2.

[0143] Comparative Example 3

[0144] (1) Shelling sorghum seeds, drying, crushing, and passing through a 100-mesh sieve to obtain sorghum flour, which is then set aside; peeling potatoes, cutting them into potato shreds, which is then set aside;

[0145] 3 g of sorghum flour, 3 g of potato shreds and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate A.

[0146] 30 mL of the above-prepared Schizophyllum Wbsh-01 bacterial solution was inoculated into the above-prepared fermentation substrate A. Based on the mass of water in the fermentation substrate A, the number of Schizophyllum Wbsh-01 inoculated per unit mass of water was 10 6 CFU / g; an aerobic fermentation culture was carried out at a temperature of 28°C for 48 hours, and an aerobic fermentation culture was carried out on an oscillator with a rotation speed of 180 rpm; a fermentation product A was obtained after an aerobic fermentation culture; the fermentation product A was sterilized at a temperature of 121°C for 30 minutes, and cooled to room temperature after sterilization to obtain material A;

[0147] (2) shelling the black beans, drying, crushing, and passing through a 100-mesh sieve to prepare black bean powder for later use; the mass percentage of crude protein in the black bean powder is 30% to 42%;

[0148] 3 g of black bean powder, 1.5 g of glucose and 300 g of deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain fermentation substrate B;

[0149] The material A obtained in step (1) and the fermentation substrate B obtained above are fully mixed, centrifuged, and the supernatant is collected.

[0150] Comparative Example 4

[0151] Shelling sorghum seeds, drying, grinding, and passing through a 100-mesh sieve to obtain sorghum flour for later use;

[0152] Peel the potatoes, cut into shredded potatoes, and set aside;

[0153] Shelling the black beans, drying, crushing, and passing through a 100-mesh sieve to prepare black bean powder for later use; the mass percentage of crude protein in the black bean powder is 30% to 42%;

[0154] 3 g sorghum flour, 3 g potato shreds, 3 g black bean flour, 1.5 g glucose and 600 g deionized water were mixed evenly, sterilized at 121° C. for 30 min, cooled to room temperature after sterilization, and set aside to obtain a fermentation substrate;

[0155] 30 mL of the Schizophyllum Wbsh-01 bacterial solution prepared in Example 1 was inoculated into the fermentation substrate prepared above, and aerobic fermentation was carried out for 15 h. The aerobic fermentation was carried out on an oscillator at a speed of 180 rpm. After aerobic fermentation, the mixture was sterilized at a temperature of 121° C. for 30 min, cooled to room temperature after sterilization, and centrifuged at a speed of 4800 rpm and a centrifugal radius of 12 cm for 30 min, and the supernatant was taken.

[0156] Comparative Example 5

[0157] Compared with Example 2, the only difference is that the inoculated bacteria in step (1) are different, 30 mL of Schizophyllum Wbsh-01 bacterial solution is replaced with 30 mL of yellow wine yeast AS2.1392 bacterial solution, and other condition parameters are the same as Example 2;

[0158] Yellow wine yeast AS2.1392 was purchased from Beijing Food Brewing Research Institute. The live bacteria concentration of yellow wine yeast AS2.1392 in the yellow wine yeast AS2.1392 bacterial solution was 10 7 CFU / mL.

[0159] Effect Example 1

[0160] The samples with soothing efficacy can significantly inhibit the increase in the expression of related inflammatory factors induced by LPS. This experiment verifies the soothing efficacy of the products prepared in the examples and comparative examples by this method.

[0161] Preparation of the test solution: The products prepared in the above examples or comparative examples were prepared into a test solution with a volume percentage of 5% using serum-free DMEM culture medium.

[0162] HaCat cells in logarithmic phase were taken and 2×10 6 The cell suspension was inoculated into a 6-well cell culture plate at a density of 1 cell / mL, 2 mL of cell suspension was added to each well, and cultured in an incubator for 24 h. After culture, the supernatant was carefully aspirated, the cells were washed twice with phosphate buffered saline (PBS), and the PBS was aspirated and discarded.

[0163] The blank group was given 2 mL of serum-free DMEM solution; the model group and the experimental group were added with 2 mL of LPS solution (LPS solution with a concentration of 200 μg / mL diluted with serum-free DMEM culture medium), and the culture was continued for 24 h.

[0164] The LPS solution with a concentration of 200 μg / mL diluted with serum-free DMEM culture medium was discarded from the model group and the experimental group, and the cells were washed twice with PBS solution. 2 mL of serum-free DMEM culture medium was added to the model group, and 2 mL of the test solution prepared above was added to the experimental group, and the cells were cultured for another 24 h.

[0165] After the culture, the cells were collected, treated with a cell lysis solution, centrifuged at 10,000 r / min and 4° C. for 10 min, and the supernatant was taken to obtain a cell lysis supernatant.

[0166] The BCA total protein content standard curve was drawn as y = 0.8795x + 0.0156, R 2 =0.9989. The standard curve of IL-8 inflammatory factor was drawn as y=0.0019x+0.0371, R 2 =0.9988.

[0167] Take 20 μL of cell lysis supernatant, use BCA kit to detect the absorbance value of the cell lysis supernatant of each group at 562 nm, and calculate the total protein content A in the cell lysis supernatant of each group using the BCA total protein content standard curve.

[0168] Take 20 μL of cell lysis supernatant, use IL-8 ELISA kit (Beijing Solebow Technology Co., Ltd.) to detect the absorbance value of cell lysis supernatant at 450 nm, use IL-8 inflammatory factor standard curve to calculate the IL-8 inflammatory factor content B in the cell lysis supernatant of each group above (remove the absorbance value when the standard concentration in the standard curve is 0 during calculation), calculate the expression amount of IL-8 inflammatory factor, the results are shown in Table 1 and Figure 2 (Table 1 and Figure 2 ***p<0.001, indicating that there is an extremely significant statistical difference compared with the model group, and it is extremely significantly reduced;** p<0.01, indicating that there is a significant statistical difference compared with the model group, and it is significantly reduced; *p<0.05, indicating that there is a statistical difference compared with the model group, and it is reduced; ns p>0.05, indicating no statistical difference compared with the model group; ### p<0.001, indicating that there is an extremely significant statistical difference compared with the blank group, extremely significantly increased).

[0169] IL-8 inflammatory factor expression = B / A. (Calculation of relative expression of inflammatory factors: First, calculate the inflammatory factor content B through the standard curve of IL-8 inflammatory factor, and divide the inflammatory factor content by the corresponding protein content A in the cell lysate. The expression of IL-8 C = B / A, and the relative expression of IL-8 = C / C 空白组 )

[0170] Table 1

[0171] OD mean Relative expression of IL-8 Blank group 0.102 1±0.086 Model Group 0.121 1.456±0.026 Example 2 0.087 0.91±0.018 Example 3 0.084 0.829±0.031 Example 4 0.084 0.846±0.036 Example 5 0.09 0.976±0.039 Example 6 0.094 1.058±0.059 Example 7 0.103 1.039±0.086 Comparative Example 1 0.113 1.429±0.039 Comparative Example 2 0.11 1.243±0.052 Comparative Example 3 0.114 1.269±0.092 Comparative Example 4 0.116 1.256±0.076 Comparative Example 5 0.116 1.231±0.047

[0172] According to the data in Table 1, after the model group was treated with LPS, the expression of the intracellular inflammatory factor IL-8 increased significantly, and the experimental group added the products prepared by Examples 2 to 7 of the present application, and the expression of the intracellular inflammatory factor IL-8 decreased, even lower than that of the blank control group, that is, the inflammatory damage caused by LPS to the cells was reduced, and the inflammatory effect was ideal. The soothing effect of the products prepared by Comparative Examples 1 to 5 was significantly worse than that of the products prepared by the examples.

[0173] Effect Example 2: DPPH free radical scavenging experiment

[0174] DPPH is an early synthesized organic free radical, which is often used to evaluate the hydrogen-donating capacity of antioxidants. It is very stable in organic solvents, purple, and has a characteristic absorption peak at 517nm. When encountering free radical scavengers, the lone pair of electrons of DPPH is paired and it fades, that is, the absorbance at the maximum absorption wavelength becomes smaller. Therefore, the scavenging effect of the sample on DPPH free radicals can be evaluated by measuring the change in absorbance.

[0175] Test solution: the product obtained in the above embodiment or comparative example;

[0176] The specific experimental steps of the DPPH free radical scavenging experiment are as follows:

[0177] (1) Take an equal volume (1 mL) of the test solution and 2 × 10 -4 mol / L DPPH solution and mix well (tube A1);

[0178] (2) Take an equal volume (1 mL) of anhydrous ethanol and 2 × 10 -4mol / L DPPH solution and mix well (tube A2);

[0179] (3) Take an equal volume (1 mL) of anhydrous ethanol and mix it with the test solution (tube A3);

[0180] (4) After 30 minutes of reaction in the dark, the absorbance of tubes A1, A2, and A3 was measured at 517 nm. The results are shown in Table 2.

[0181] The calculation formula of DPPH free radical scavenging rate is: DPPH free radical scavenging rate = [(A2 + A3) - A1] / A2 × 100%, the calculation results are shown in Table 2 and Figure 3 (Table 2 and Figure 3 ***p<0.001, indicating that there is an extremely significant statistical difference compared with Example 2, and the decrease is extremely significant; ** p<0.01, indicating that there is a significant statistical difference compared with Example 2, and it is significantly reduced; *p<0.05, indicating that there is a statistical difference compared with Example 2, and it is reduced; ns p>0.05, indicating no statistical difference compared with Example 2).

[0182] Table 2

[0183]

[0184]

[0185] The results in Table 2 show that the DPPH radical scavenging ability of the products prepared in Examples 2 to 7 of the present application is significantly better than that of the products prepared in Comparative Examples 1 to 5. The type of fermentation substrate, fermentation strain, fermentation culture time and step-by-step fermentation process have a great influence on the antioxidant efficacy of the prepared products.

[0186] Effect Example 3 Hydroxyl Radical Scavenging Experiment

[0187] The Fenton reaction is used to generate hydroxyl radicals (·OH), and salicylic acid is added to the reaction system. ·OH reacts with salicylic acid to generate a colored compound 2,3-dihydroxybenzoic acid with characteristic absorption at 510nm. The fixed reaction time method is used to measure the absorbance of the reaction solution containing the analyte at 510nm, and compared with the blank solution to determine the scavenging effect of the analyte on ·OH.

[0188] The liquid to be tested is the product obtained in the above-mentioned embodiment and comparative example;

[0189] Add reagents to the test tube according to Table 3, and add 9mmol / L FeSO 4 , 9mmol / L salicylic acid ethanol solution, test solution, appropriate amount of deionized water and 8.8mmol / L H 2 O2 Solution. Shake well, heat in a 37℃ water bath for 15 minutes, take out, and measure the absorbance of the blank control A 0 , sample group absorbance A x and without H 2 O 2 The background absorbance of the solution is A x0 . Determination of A 0 The reference solution was a system without hydrogen peroxide. Three parallel experiments were performed in each group, and the absorbance values ​​were tested and the average values ​​were taken. The results are shown in Table 4. The hydroxyl radical scavenging rate of each group was calculated according to the following formula. The results are shown in Table 4 and Figure 4 (Table 4 and Figure 4 ***p<0.001, indicating that there is an extremely significant statistical difference compared with Example 2, and the decrease is extremely significant; ** p<0.01, indicating a statistically significant difference compared with Example 2, which is significantly reduced; ns p>0.05, indicating no statistical difference compared with Example 2).

[0190] Hydroxyl free radical scavenging rate = (A 0 -(A x -A x0 )) / A 0 ×100%.

[0191] Table 3

[0192]

[0193]

[0194] Table 4

[0195]

[0196] The results in Table 4 show that the hydroxyl radical scavenging ability of the products prepared in Examples 2 to 7 of the present application is significantly better than that of the products prepared in Comparative Examples 1 to 5. The type of fermentation substrate, fermentation strain, fermentation culture time and step-by-step fermentation process have a great influence on the antioxidant effect of the prepared product.

[0197] Effect Example 4: Anti-light damage

[0198] 1. Experimental steps:

[0199] The products obtained in the above examples and comparative examples were respectively prepared with serum-free DMEM culture medium to prepare test solutions of the experimental group with a volume percentage of 10%.

[0200] HaCaT cells were cultured in a medium containing 10% fetal bovine serum and 1% double antibody (1×10 5The cells were grown in DMEM medium containing 100 U / L penicillin and 100 mg / L streptomycin. 2 In a saturated humidity incubator, when the cell confluence reaches more than 85%, the cells in the logarithmic growth phase are digested with 0.05% trypsin, and the digestion reaction is terminated with serum-containing DMEM. The cell count plate is used to count and the cell suspension concentration is adjusted to 7×10 4 The cell suspension was inoculated into a 96-well plate at a ratio of 100 μL per well and incubated at 37°C and 5% CO 2 After incubation, the old culture medium was removed, the cells were washed twice with phosphate buffer, and then 100 μL of PBS solution was added. The model group and the experimental group were irradiated with UVB at a dose of 15 mJ / cm 2 The negative control group was not irradiated; PBS was discarded, serum-free DMEM medium was added to the model group and the negative control group, 100 μL of the above-prepared filtered and sterilized experimental group test solution was added to each well of the experimental group, and 3 replicates were made for each test solution; the blank control group had no cells and 100 μL of PBS was added. Then, the cells were incubated at 37°C and 5% CO 2 Then, 10 μL of CCK-8 solution was added to each well and incubated for another 3 h. The absorbance value was measured at a wavelength of 450 nm and the cell survival rate of each group was calculated. The results are shown in Table 5 and Figure 5 (Table 5 and Figure 5 ***p<0.001, indicating that there is an extremely significant statistical difference compared with the model group, and it is extremely significantly increased; ** p<0.01, indicating that there is a significant statistical difference compared with the model group, and it is significantly increased; *p<0.05, indicating that there is a statistical difference compared with the model group, and it is increased; ns p>0.05, indicating no statistical difference compared with the model group; ### p<0.001, indicating that there is an extremely significant statistical difference compared with the blank group, extremely significantly reduced).

[0201] The cell viability was calculated as follows:

[0202] Cell survival rate (%) = (A experimental group or model group - A blank control group) / (A negative control group - A blank control group) × 100%.

[0203] Table 5

[0204]

[0205]

[0206] According to the results in Table 5, the cell survival rate of the model group was significantly reduced after ultraviolet stimulation. The products prepared in Examples 2 to 7 of the present application have ideal repair effects on damaged cells after ultraviolet stimulation, can improve cell survival ability, and the repair effect is significantly better than the products prepared in the comparative example.

[0207] Effect Example 5: Human skin fibroblast toxicity experiment

[0208] This experiment uses human skin fibroblasts from the Chinese Science Cell Bank to verify the cytotoxicity of the products obtained in the above examples and comparative examples.

[0209] Reagents: 0.25% (containing EDTA) trypsin was produced by GIBCO, USA; DMEM culture medium was produced by GIBCO, USA; double antibody was produced by Corning, USA; CCK-8 was produced by Beijing Biode Biotech Co., Ltd.; fetal bovine serum was produced by GIBCO, USA; phosphate buffer was produced by Beijing Bioregi Biotech Co., Ltd.

[0210] Equipment: WJ-80A-Ⅱ CO 2 The manufacturer of the constant temperature incubator is Shanghai Shengke Instrument Equipment Co., Ltd.; the manufacturer of the Sunrise microplate reader is Tecan Trading Co., Ltd.; the manufacturer of the TL80-2 medical centrifuge is Jiangsu Tianli Medical Equipment Co., Ltd.; the manufacturer of the NUNC 96-well cell culture plate is Thermo Fisher Scientific.

[0211] 1. Experimental steps:

[0212] The products prepared in the above-mentioned examples and comparative examples were respectively prepared into 5% by volume of the test solution of the experimental group using serum-free DMEM culture medium.

[0213] Human skin fibroblasts were cultured in a medium containing 10% fetal bovine serum and 1% double antibody (1×10 5 The cells were grown in DMEM medium containing 100 U / L penicillin and 100 mg / L streptomycin. 2 In a saturated humidity incubator, when the cell confluence reaches more than 85%, the cells in the logarithmic growth phase are digested with 0.05% trypsin, and the digestion reaction is terminated with serum-containing DMEM. The cell count plate is used to count and the cell suspension concentration is adjusted to 7×10 4 The cell suspension was inoculated into a 96-well plate at a ratio of 100 μL per well and incubated at 37°C and 5% CO 2Incubate for 12 hours under the same conditions. Remove the old culture medium and wash the cells twice with phosphate buffer. Add 100 μL of the above-prepared sterilized test solution of different concentrations to each well of the experimental group, and make 6 duplicate wells for each test solution; the control group contains cells and serum-free DMEM medium is added; the blank control group has no cells and 100 μL of PBS is added. Then incubate at 37°C and 5% CO 2 Then, 10 μL of CCK-8 solution was added to each well and incubated for another 3 h. The absorbance was measured at 450 nm and the cell survival rate of each group was calculated. The results are shown in Table 6 and Figure 6 (Table 6 and Figure 6 ***p<0.001, indicating that there is an extremely significant statistical difference compared with Example 2, and the decrease is extremely significant; ** p<0.01, indicating a statistically significant difference compared with Example 2, which is significantly reduced; ns p>0.05, indicating no statistical difference compared with Example 2).

[0214] The cell viability was calculated as follows:

[0215] Cell survival rate = (A experimental group - A blank control group) / (A control group - A blank control group) × 100%.

[0216] Table 6

[0217]

[0218] The results show that the products prepared in Examples 2 to 7 of the present application are not only non-cytotoxic, but some of them also have an ideal effect of promoting cell growth. It can be seen that they have an ideal anti-aging effect.

[0219] Finally, it should be noted that in this application, the terms "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also other elements that are not explicitly listed, or also includes elements that are inherent to such process, method, article or apparatus.

[0220] Although the present application has been disclosed above through the description of the specific embodiments of the present application, it should be understood that those skilled in the art can design various modifications, improvements or equivalents to the present application within the spirit and scope of the attached scheme. These modifications, improvements or equivalents should also be considered to be included in the scope of protection claimed in the present application.

Claims

1. A method for preparing a white ginseng fermentation product having a soothing effect, characterized in that: The steps include: (1) The fermentation substrate A includes 100 parts of water, 0.4-5 parts of carbohydrate carbon source and 0.5-5 parts of organic nitrogen source; Schizophyllum commune ) is inoculated into the fermentation substrate A, and subjected to aerobic fermentation culture for 24 to 48 hours to obtain a fermentation product A; (2) The fermentation substrate B includes 100 parts of water, 0.4-2 parts of a carbohydrate carbon source and 0.5-2 parts of an organic nitrogen source; the fermentation product A is mixed with the fermentation substrate B, and the mixture is subjected to secondary aerobic fermentation for 12-20 hours, and sterilized to obtain the white ginseng fungus fermentation liquid; Wherein, in step (1), the white ginseng fungus is Schizophyllum commune Wbsh-01 (Schizophyllum commune) preserved in the General Microbiological Center of China Microorganism Culture Collection Committee, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing 100101, preservation date is March 9, 2023, and preservation number is CGMCC No.40388; the carbohydrate carbon source includes monosaccharides and / or starch-containing materials, the monosaccharides include glucose, and the starch-containing materials include at least one of rice flour, sorghum flour, corn flour and cassava starch; the organic nitrogen source includes at least one of potato, soybean flour, black soybean flour, soybean meal and soybean peptide; based on the water in the fermentation substrate A, the number of the white ginseng fungus inoculated per unit mass of the water is 10 5 CFU / g~10 7 CFU / g; In step (2), the carbohydrate carbon source includes monosaccharides and / or starch-containing materials, the starch-containing materials include at least one of rice flour, sorghum flour, corn flour and cassava starch, and the organic nitrogen source includes at least one of potato, soybean flour, black soybean flour, soybean meal and soybean peptide.

2. The method for preparing the white ginseng fungus fermentation product with soothing effect according to claim 1, characterized in that: The preparation method satisfies at least one of the following conditions: In step (1), the water includes deionized water; In step (1), the weight proportion of the carbohydrate carbon source is 0.5 to 2 parts; In step (1), the weight portion of the organic nitrogen source is 0.5 to 2 parts; In step (1), the fermentation substrate A also includes a sterilization operation before use; In step (1), the white ginseng fungus is added in the form of a white ginseng fungus liquid; In step (1), the time of the aerobic fermentation culture is 35 to 48 hours; In step (1), the temperature of the primary aerobic fermentation culture is 25-35°C; In step (1), the primary aerobic fermentation culture is carried out under shaking conditions, and the shaking speed is 150-250 rpm.

3. The method for preparing the white ginseng fermentation product with soothing effect according to claim 2, characterized in that: The preparation method satisfies at least one of the following conditions: In step (1), the mass percentage of crude protein in the black soybean powder is 30% to 42% of the black soybean powder; In step (1), the weight proportion of the carbohydrate carbon source is 1 to 1.7 parts; In step (1), the weight proportion of the organic nitrogen source is 0.8 to 1.5 parts; In step (1), the concentration of live bacteria of the white ginseng fungus in the white ginseng fungus liquid is 10 6 ~10 8 CFU / mL; In step (1), the temperature of the primary aerobic fermentation culture is 26-30°C; In step (1), the primary aerobic fermentation culture is carried out under shaking conditions, and the shaking speed is 170-200 rpm.

4. The method for preparing the white ginseng fungus fermentation product with soothing effect according to any one of claims 1 to 3, characterized in that: The preparation method satisfies at least one of the following conditions: In step (2), the water includes deionized water; In step (2), the weight percentage of the carbohydrate carbon source is 0.5 to 1.5 parts; In step (2), the weight portion of the organic nitrogen source is 0.8 to 2 parts; In step (2), the fermentation substrate B also includes a sterilization operation before use; In step (2), the secondary aerobic fermentation culture time is 12 to 16 hours; In step (2), the temperature of the secondary aerobic fermentation culture is 25-35°C; In step (2), the secondary fermentation culture is carried out under shaking conditions, and the shaking speed is 150-250 rpm; In step (2), the sterilization method is high temperature sterilization.

5. The method for preparing the white ginseng fungus fermentation product with soothing effect according to claim 4, characterized in that: The preparation method satisfies: In step (2), the sterilization operation also includes centrifugation and collection of the supernatant and / or mixing with a preservative.

6. The method for preparing the white ginseng fermentation product with soothing effect according to claim 5, characterized in that: The preparation method satisfies at least one of the following conditions: In step (2), the monosaccharide includes glucose; In step (2), the mass percentage of crude protein in the black bean powder is 30% to 42% of the black bean powder; In step (2), the weight percentage of the carbohydrate carbon source is 0.5 to 0.7 parts; In step (2), the weight percentage of the organic nitrogen source is 0.8 to 1.67 parts; In step (2), the temperature of the secondary aerobic fermentation culture is 26-30°C; In step (2), the secondary fermentation culture is carried out under shaking conditions, and the shaking speed is 160-200 rpm; In step (2), the centrifugal speed is 3000-9000 rpm; In step (2), the centrifugation time is 10 to 40 minutes; In step (2), the radius of the centrifuge is 8 to 15 cm; In step (2), during the mixing process with the preservative, the mixing temperature is 60-80°C; In step (2), in the process of mixing with the preservative, the preservative used includes p-hydroxyacetophenone and / or 1,2-hexanediol.

7. The method for preparing the white ginseng fermentation product with soothing effect according to claim 6, characterized in that: The preparation method satisfies at least one of the following conditions: In step (2), after the sterilization operation of the fermentation substrate B, the step further includes cooling the fermentation substrate B to room temperature; In step (2), the centrifugal speed is 4000-6000 rpm; In step (2), the centrifugation time is 20 to 40 minutes; In step (2), during the mixing process with the preservative, the mixing temperature is 70-80°C; In step (2), the mass percentage of the p-hydroxyacetophenone in the material obtained after sterilization is 0.3% to 1%; the mass percentage of the 1,2-hexanediol in the material obtained after sterilization is 0.3% to 1%.

8. The method for preparing the white ginseng fermentation product with soothing effect according to claim 7, characterized in that: The preparation method satisfies at least one of the following conditions: In step (2), the mass percentage of the p-hydroxyacetophenone in the material obtained after sterilization is 0.5%; the mass percentage of the 1,2-hexanediol in the material obtained after sterilization is 0.5%.

9. A white ginseng fermentation product with soothing effect, characterized in that: The product is prepared by the method for preparing the white ginseng fungus fermentation product with soothing effect as described in any one of claims 1 to 8.

10. Use of the soothing effect of the white ginseng fermentation product as claimed in claim 9 directly as a product, as an additive or as a base in the preparation of a skin external preparation.

11. The use according to claim 10, characterized in that The soothing effect of the white ginseng fermentation product is used as at least one of the anti-inflammatory active ingredient, antioxidant active ingredient, skin repair active ingredient and anti-aging active ingredient in the skin external preparation.

12. A skin external preparation, characterized in that: The invention comprises the white ginseng fungus fermentation product with soothing effect as claimed in claim 9.

13. The external skin preparation according to claim 12, wherein The skin topical preparation satisfies at least one of the following conditions: The skin external preparation also includes an active ingredient; The skin external preparation includes a facial mask, essence or toner; The mass percentage of the white ginseng fermentation product with soothing effect in the skin external preparation is 5% to 99%.

14. The external skin preparation according to claim 13, wherein The skin topical preparation satisfies at least one of the following conditions: The active ingredient comprises at least one of a moisturizing active ingredient, a whitening active ingredient, an anti-inflammatory active ingredient, an anti-allergic active ingredient and an antioxidant active ingredient; The mass percentage of the white ginseng fermentation product with soothing effect in the skin external preparation is 60% to 99%.

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