A red ginseng fermentation liquor, a preparation method thereof and an application thereof

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a red ginseng fermentation liquor and a preparation method and application thereof. The preparation method of the red ginseng fermentation liquor comprises the following steps: dissolving red ginseng powder in water to obtain a red ginseng fermentation culture medium by destroying the cell wall of the red ginseng; mixing seed liquid of plant lactobacillus and seed liquid of fermented mucous lactobacillus to obtain a composite strain seed liquid; inoculating the composite strain seed liquid into the red ginseng fermentation culture medium for fermentation culture to obtain a fermentation liquor; centrifuging the fermentation liquor, collecting supernatant, and extruding and filtering the supernatant to collect filtrate, and the obtained filtrate is the red ginseng fermentation liquor. The application adopts composite strain fermentation and combines a mechanical extrusion mode to improve the content of total ginsenosides. Through the method, the concentration of total ginsenosides is increased from 2.19 mg / mL to 7.53 mg / mL.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a red ginseng fermentation liquor and a preparation method and application thereof. BACKGROUND

[0002] Ginseng belongs to the Panax genus of the Araliaceae family, and has extensive and good medicinal and health-care functions in cardiovascular system diseases, mental system diseases, digestive system diseases, hematopoietic and immune system diseases, endocrine system diseases, anti-tumor, anti-aging, tonifying qi and blood, hair growth and hair loss prevention, etc. Red ginseng is a cooked product of ginseng, which is steamed and dried, and has a function of being more warm and nourishing. Red ginseng not only retains various effective components of ginseng, but also has better effects of tonifying qi and cultivating essence, nourishing blood and producing fluid, strengthening heart and stomach, etc. Compared with ginseng, the performance of red ginseng is more moderate. The content of ginsenosides in ginseng accounts for about 4% of the effective components, which plays a significant role in medicine. Among them, the contents of natural prototype ginsenosides Rb1, Rb2, Rc, Re and Rg1 account for 70% to 80% of the total content of ginsenosides; and the contents of rare ginsenosides Rg3, Rh1, Rh2, F2 and CK are extremely low, but have high pharmacological activities. For example, rare ginsenoside Rg3 is considered to be a comprehensive regulator of human functions, and has effects of anti-inflammation, anti-cancer, protection of cardiovascular system, improvement of cognition, protection of nerves and immune regulation. Rare ginsenoside F2 is one of the core functional components of red ginseng, and has multiple effects of anti-oxidation, anti-photoaging, wrinkle reduction, promotion of skin hydration and repair of skin barrier. A large number of studies have shown that natural prototype ginsenosides can be converted into rare ginsenosides through intestinal microbial flora metabolism in vivo, which improves the pharmacological activity. At the same time, rare ginsenosides can easily cross the blood-brain barrier with small molecular weight and strong liposolubility, and directly reach the brain, which further improves the absorption efficiency and bioavailability in vivo, and opens up new possibilities for the treatment and health-care field.

[0003] As a key active ingredient in ginseng, ginsenosides have a wide application prospect in the fields of medicine, health-care products and cosmetics. However, the current preparation process of ginsenosides is faced with the key problem of low yield, which seriously restricts the large-scale production and application of ginsenosides.

[0004] In the existing microbial fermentation technology, the utilization efficiency of ginseng raw materials by microorganisms is generally low. On the one hand, complex chemical components in ginseng, such as polysaccharides and proteins, may inhibit the growth and metabolic activity of microorganisms, so that the microorganisms cannot fully exert the conversion capacity of ginsenosides. On the other hand, the growth characteristics and metabolic regulation mechanism of microorganisms also limit the yield of ginsenosides. For example, the conversion enzyme activity of some microorganisms to ginsenosides is not high during the fermentation process, resulting in low substrate conversion rate; or the growth of microorganisms is inhibited in the late growth stage due to the consumption of nutrients and accumulation of metabolic products, which further affects the continuous synthesis of ginsenosides.

[0005] Due to the low yield of ginsenosides prepared by microbial fermentation, the production cost is high, which makes the price of ginsenosides related products expensive, and it is difficult to meet the market demand. This not only hinders the wide application of ginsenosides in the medical field, but also limits its large-scale promotion in the health care and cosmetic industries. Therefore, it is urgent to develop a method that can significantly improve the yield of ginsenosides prepared by microbial fermentation. SUMMARY

[0006] In order to solve the problem of low yield of ginsenosides prepared by microbial fermentation, the present application provides a preparation method of red ginseng fermentation broth.

[0007] The technical scheme adopted by the present application is:

[0008] The first aspect of the present application provides a preparation method of red ginseng fermentation broth, comprising the following steps:

[0009] After dissolving the red ginseng powder in water, cellulase and pectinase are added to destroy the cell wall in the red ginseng, and then the enzyme is inactivated and sterilized to obtain a red ginseng fermentation medium; a seed liquid of lactobacillus plantarum is prepared; a seed liquid of lactobacillus fermentum is prepared; the seed liquid of lactobacillus plantarum and the seed liquid of lactobacillus fermentum are mixed to obtain a composite strain seed liquid; the composite strain seed liquid is inoculated into the red ginseng fermentation medium for fermentation culture to obtain a fermentation broth; the fermentation broth is centrifuged, the supernatant is collected, and the supernatant is squeezed and filtered to collect the filtrate, and the obtained filtrate is the red ginseng fermentation broth.

[0010] Preferably, the pore size of the filter membrane used in the squeezing and filtering is 0.8 μm.

[0011] Preferably, the volume ratio of the seed liquid of lactobacillus plantarum to the seed liquid of lactobacillus fermentum is 1:1, and the inoculation amount of the composite strain seed liquid is 4% of the volume fraction.

[0012] Preferably, the number of viable bacteria of lactobacillus plantarum in the seed liquid of lactobacillus plantarum is 1×10 8 CFU / mL; and the number of viable bacteria of lactobacillus fermentum in the seed liquid of lactobacillus fermentum is 1×10 8 CFU / mL.

[0013] Preferably, the addition amount of cellulase and pectinase is: 0.05 mL of cellulase and 0.05 mL of pectinase are added per 1 g of red ginseng powder.

[0014] Preferably, the fermentation culture condition is 37℃ static culture for 48h.

[0015] The second aspect of the present application provides a red ginseng fermentation broth obtained according to the preparation method.

[0016] The third aspect of the present application provides a use of the red ginseng fermentation liquor for preparing a product for improving immunity.

[0017] Preferably, the product further comprises a pharmaceutically acceptable adjuvant, which is a sweetener.

[0018] Preferably, the sweetener is xylitol.

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

[0020] The present application provides a preparation method of red ginseng fermentation liquor, comprising the following steps:

[0021] The red ginseng powder is dissolved in water, and then cellulase and pectinase are added to destroy the cell wall in the red ginseng, followed by enzyme inactivation, sterilization, to obtain a red ginseng fermentation culture medium; a seed liquid of Lactobacillus plantarum is prepared; a seed liquid of Lactobacillus fermentum is prepared; the seed liquid of Lactobacillus plantarum and the seed liquid of Lactobacillus fermentum are mixed to obtain a composite strain seed liquid; the composite strain seed liquid is inoculated into the red ginseng fermentation culture medium for fermentation culture, to obtain a fermentation liquor; the fermentation liquor is centrifuged, the supernatant is collected, and the supernatant is squeezed and filtered, and the filtrate is collected, which is the red ginseng fermentation liquor. The present application uses composite strain fermentation and combines mechanical squeezing to improve the content of total ginsenosides. Through the method of the present application, the concentration of total ginsenosides is increased from 2.19 mg / mL to 7.53 mg / mL.

[0022] Lactobacillus plantarum and Lactobacillus fermentum, as probiotics, can spontaneously produce bacterial outer vesicles during fermentation. The present application collects the supernatant of the fermentation liquor fermented by the composite strain, and then prepares a red ginseng fermentation liquor containing bacterial outer vesicles through mechanical squeezing. The bacterial outer vesicles can improve the health status of the intestinal flora of the human body, and can effectively encapsulate total ginsenosides to improve the storage stability thereof. In addition, the red ginseng fermentation liquor of the present application has anti-inflammatory and antioxidant effects.

[0023] The present application uses red ginseng as a substrate for cultivation. On the one hand, probiotics can produce bacterial outer vesicles during growth, and on the other hand, the growth and metabolism of probiotics can transform natural prototype ginsenosides in red ginseng to produce rare ginsenosides and other secondary metabolites with high physiological activity. During mechanical squeezing, bacterial outer vesicles can enrich and encapsulate rare ginsenosides, which can be applied in the field of food and beverage and cosmetic efficacy raw materials. This method simplifies the process of engineering bacterial outer vesicles to encapsulate rare ginsenosides, and can effectively improve the problems of poor water solubility and poor stability of rare ginsenosides.

[0024] The present application combines probiotic fermentation technology and bacterial outer vesicle preparation technology, effectively solves the problem that probiotics only have the function of fermentation agent, and the present application also overcomes the problem of poor stability of rare saponins in aqueous solution. The system of bacterial outer vesicles encapsulating rare saponins to form nanoemulsions can significantly improve the stability of rare saponins and the ability to resist gastric acid. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 A and B are bacterial outer vesicles in filtrate through 0.8 μm filter membrane and 0.1 μm filter membrane, respectively.

[0026] Figure 2 A and B are particle size distribution diagrams of bacterial outer vesicles in filtrate through 0.8 μm filter membrane and 0.1 μm filter membrane, respectively.

[0027] Figure 3 The figure is the concentration of total ginsenosides in red ginseng fermentation broth prepared in Example 1 and Comparative Example 1.

[0028] Figure 4 A and B are transmission electron microscope photos of bacterial outer vesicle structure after in vitro digestion simulation.

[0029] Figure 5 A, B, C and D are bacterial outer vesicle structure characterization under different storage conditions. DETAILED DESCRIPTION

[0030] The present application will be further described below through specific examples, but the scope of the present application is not limited thereto. The details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and these modifications or replacements all fall within the protection scope of the present application.

[0031] The inventive concept of the present application is as follows:

[0032] In the existing microbial fermentation technology, the utilization efficiency of ginseng raw materials by microorganisms is generally low, the main reasons are: the complex chemical components in ginseng can inhibit the growth and metabolic activity of microorganisms, so that the microorganisms cannot fully exert the transformation ability of total ginsenosides, and the growth characteristics and metabolic regulation mechanism of the organism also limit the yield of total ginsenosides.

[0033] Based on this, the present application provides a preparation method of red ginseng fermentation broth, comprising the following steps:

[0034] The red ginseng powder is dissolved in water, cellulase and pectinase are added to destroy the cell wall in the red ginseng, then the enzyme is inactivated, sterilized, and a red ginseng fermentation culture medium is obtained; a seed liquid of lactobacillus plantarum is prepared; a seed liquid of lactobacillus fermentum is prepared; the seed liquid of lactobacillus plantarum and the seed liquid of lactobacillus fermentum are mixed to obtain a compound strain seed liquid; the compound strain seed liquid is inoculated into the red ginseng fermentation culture medium for fermentation culture, and a fermentation liquid is obtained; the fermentation liquid is centrifuged, the supernatant is collected, and the supernatant is squeezed and filtered, and the filtrate is collected, and the obtained filtrate is the red ginseng fermentation liquid.

[0035] As probiotics, lactobacillus plantarum and lactobacillus fermentum can degrade the cell wall of red ginseng through cellulase and hemicellulase in the probiotics, so as to release natural prototype saponins in the red ginseng cells; on the other hand, glycosidase in the probiotics, especially beta-glucosidase, can hydrolyze the glycosidic bond in the natural prototype saponins, so as to convert them into rare saponins, thereby improving the content of rare saponins.

[0036] The probiotics can spontaneously produce BEVs, which are spherical structures with a lipid bilayer, generally with a diameter of 20nm-400nm. As bacterial-derived small-molecule active substances, BEVs have advantages such as stable structure and long effective circulation time, and can encapsulate various bioactive molecules, including proteins, lipids, nucleic acids and metabolites. A large number of studies have shown that BEVs have potential mechanisms in immune response, antioxidant and other aspects, and have application potential as new postbiotics in functional foods, disease treatment substitutes and immune regulation adjuvants. In addition, BEVs are also ideal carriers for nano-delivery systems of bioactive compounds, because they have good immunological properties and pharmacokinetic properties, and have the ability to penetrate physiological barriers, so they can be loaded with high-value functional ingredients through technical means to prepare delivery systems. After taking related products, precise targeted nutritional intervention can be achieved, which can be used as an important carrier for constructing food nutrition factor delivery systems.

[0037] In order to enable those skilled in the art to better understand the technical solutions of the present application and to implement them, the present application will be further described below in conjunction with specific embodiments. In the description of the present application, if not specially stated, the reagents used are commercially available, and the methods used are conventional techniques in the art.

[0038] The abbreviations used in the present application are shown in Table 1.

[0039] Table 1 Abbreviation Table

[0040]

[0041] The strains and manufacturer information used in the present application are shown in Table 2.

[0042] Table 2 Strain and Manufacturer Information

[0043]

[0044] The cellulase and pectinase used in the application are purchased from Novozymes, the enzyme activity of the cellulase is 700 U / mL, and the enzyme activity of the pectinase is 10,000 U / mL; the red ginseng powder is purchased from Jilin Hongjiu Biological Technology Co., Ltd.; the artificial saliva, artificial gastric juice and artificial intestinal juice are purchased from Shanghai Xinfan Biological Technology Co., Ltd.

[0045] In the application, the detection method of the total ginsenoside content is as follows: 20 μL of the red ginseng fermentation liquor is added into 5 g / L 0.2 mL of a vanillin solution and mixed, 0.8 mL of an analytical pure perchloric acid is added and mixed, 10 min of constant temperature water bath is carried out at 60 DEG C, and after cooling on ice, glacial acetic acid is added to 5 mL and shaken up. The absorbance is measured at a wavelength of 560 nm.

[0046] Example 1

[0047] A preparation method of the red ginseng fermentation liquor is as follows:

[0048] S1, the red ginseng powder is dissolved in water, the cell wall in the red ginseng is destroyed by adding cellulase and pectinase, then the enzyme is inactivated, sterilized, and the red ginseng fermentation medium is obtained.

[0049] The red ginseng powder is mixed with distilled water at a mass ratio of 1:10, the red ginseng powder is fully dissolved, then the cellulase and pectinase are added; the enzyme is hydrolyzed at 60 DEG C with a shaking speed of 100 rpm / min for 3 h; the enzyme is inactivated at 85 DEG C for 10 min, then sterilized at 121 DEG C for 20 min, and cooled to room temperature to obtain the red ginseng fermentation medium. The cellulase and pectinase are added in an amount of 0.05 mL of cellulase and 0.05 mL of pectinase per 1 g of red ginseng powder.

[0050] S2, the seed liquid of the lactobacillus plantarum and the seed liquid of the lactobacillus fermentum are prepared respectively, the seed liquid of the lactobacillus plantarum and the seed liquid of the lactobacillus fermentum are mixed to obtain the seed liquid of the composite strain.

[0051] The YZ-lactobacillus plantarum LP28 is dissolved in sterile water to obtain the YZ-lactobacillus plantarum LP28 bacterial liquid, the YZ-lactobacillus plantarum LP28 bacterial liquid is gently streaked on the MRS solid culture medium using an inoculation loop, and cultured at 37 DEG C until the single colony of the YZ-lactobacillus plantarum LP28 appears. The single colony of the YZ-lactobacillus plantarum LP28 is picked up to the test tube containing 5 mL of the sterilized MRS liquid medium using an inoculation loop, and the strain is activated, and cultured at 37 DEG C for 18 h, to obtain the seed liquid of the YZ-lactobacillus plantarum LP28, and the viable bacterial count of the YZ-lactobacillus plantarum LP28 in the seed liquid of the YZ-lactobacillus plantarum LP28 is 1 x 10 8CFU / mL.

[0052] The fermentation Lactobacillus muciadis JYLF-71 was dissolved in sterile water to obtain a fermentation Lactobacillus muciadis JYLF-71 liquid, and the fermentation Lactobacillus muciadis JYLF-71 liquid was streaked on MRS solid medium with an inoculation loop, and cultured at 37°C until single colonies of the fermentation Lactobacillus muciadis JYLF-71 appeared. The single colonies of the fermentation Lactobacillus muciadis JYLF-71 were picked up with an inoculation loop into a test tube containing 5 mL of sterilized MRS liquid medium, and the strain was activated and cultured at 37°C for 18 h. The seed liquid of the fermentation Lactobacillus muciadis JYLF-71 was obtained, and the viable cell count of the fermentation Lactobacillus muciadis JYLF-71 in the seed liquid was 1×10 8 CFU / mL.

[0053] The seed liquid of the YZ-Lactobacillus plantarum LP28 and the seed liquid of the fermentation Lactobacillus muciadis JYLF-71 were mixed in a volume ratio of 1:1 to obtain a composite strain seed liquid.

[0054] S3, the composite strain seed liquid was inoculated into the red ginseng fermentation medium for fermentation culture to obtain a fermentation liquid.

[0055] The composite strain seed liquid was inoculated into the red ginseng fermentation medium at a volume fraction of 4%, and cultured at 37°C for 48 h in an incubator. The fermentation was completed, and the fermentation liquid was collected.

[0056] S4, the fermentation liquid was centrifuged, and the supernatant was collected. The supernatant was squeezed and filtered, and the filtrate was collected. The obtained filtrate was the red ginseng fermentation liquid.

[0057] The fermentation liquid was centrifuged at 4000 rpm / min for 5 min at room temperature to remove the powder of the red ginseng powder in the fermentation liquid, and the supernatant in the fermentation liquid was retained. Since the YZ-Lactobacillus plantarum LP28 and the fermentation Lactobacillus muciadis JYLF-71 can spontaneously produce bacterial outer vesicles during fermentation, the following operations were performed on the supernatant:

[0058] The supernatant was sequentially passed through filter membranes with pore sizes of 8 μm, 5 μm, 1 μm, 0.8 μm and 0.1 μm, and at the same time, a hand-push liposome extruder was used to squeeze the supernatant when it passed through each filter membrane. The supernatant passing through each filter membrane was unidirectionally squeezed for 6 times to obtain the filtrate under the corresponding pore size, and 50 mL of the filtrate under each pore size was collected for subsequent detection. The remaining filtrate was repeatedly squeezed after replacing the next filter membrane until the supernatant was completely passed through the 0.1 μm filter membrane. The filtrate passing through the 0.8 μm filter membrane was the red ginseng fermentation liquid.

[0059] Comparative Example 1

[0060] A preparation method of a red ginseng fermentation liquid is as follows:

[0061] S1 to S3 are exactly the same as those in Example 1. In S4, after the fermentation is completed, the fermentation liquid is centrifuged at 4000 rpm / min for 5 min at room temperature to remove the red ginseng powder in the fermentation liquid. The supernatant in the fermentation liquid is the red ginseng fermentation liquid.

[0062] The filtrate passed through the filter membranes of various pore sizes in Example 1 was characterized as follows:

[0063] The filtrates passed through 8μm, 5μm, 1μm, 0.8μm, and 0.1μm filters were examined for the presence of the Tyndall effect. The filtrates passed through 0.8μm and 0.1μm filters were then observed using transmission electron microscopy, and the laser particle size distribution and protein concentration were measured.

[0064] Transmission electron microscopy results Figure 1 The particle size analysis results are shown in Figure 2 As shown. Figure 1 It can be seen that the bacterial extracellular vesicles in the filtrate are teacup-shaped structures. Figure 2 It can be seen that the average particle size of bacterial extracellular vesicles in the filtrate passed through a 0.8 μm filter membrane is 147 nm, and the average particle size of bacterial extracellular vesicles in the filtrate passed through a 0.1 μm filter membrane is 132.4 nm.

[0065] In addition, the vesicle concentration and protein concentration of bacterial extracellular vesicles in the filtrate were determined. The test results showed that the concentration of bacterial extracellular vesicles in the filtrate after passing through the 0.8 μm filter membrane was 1.8×10 10 Particles / mL, protein concentration was 7.30 mg / mL; the concentration of bacterial extracellular vesicles in the filtrate after passing through a 0.1 μm filter membrane was 1.4×10 10 Particles / mL, and the protein concentration was 6.24 mg / mL.

[0066] The total ginsenosides in the red ginseng fermentation broth prepared in Example 1 and Comparative Example 1 were determined as follows:

[0067] Depend on Figure 3 It can be seen that the total ginsenoside concentration in the red ginseng fermented liquid prepared in Comparative Example 1 was 2.19 mg / mL, while the total ginsenoside concentration in the red ginseng fermented liquid prepared in Example 1 was increased to 7.53 mg / mL. This shows that the total ginsenoside concentration was significantly increased after extrusion.

[0068] In addition, 20% SDS of equal volume mass fraction was added to the red ginseng fermentation broth prepared in Example 1 for demulsification. After demulsification, the concentration of total ginsenosides reached 9.55 mg / mL, indicating that bacterial extracellular vesicles can effectively encapsulate total ginsenosides.

[0069] Example 2

[0070] An application of a red ginseng fermentation liquor, specifically as follows:

[0071] 1. Immunity effect of the red ginseng fermentation liquor.

[0072] Taking macrophage Raw264.7 as the research object, the cells were treated with the red ginseng fermentation liquor prepared in Example 1, and the change of cell activity was observed and counted to evaluate the immunity effect. The grouping and treatment of the cell proliferation test are shown in Table 3.

[0073] Table 3 Grouping and treatment of cell proliferation test

[0074]

[0075] Taking macrophage Raw264.7 as the research object, the cells were treated with the red ginseng fermentation liquor prepared in Example 1, and the change of cell activity was observed and counted to evaluate the immunity effect. The grouping and treatment of the cell proliferation test are shown in Table 3.

[0076] Table 4 Grouping and treatment of cell phagocytosis test

[0077]

[0078] Note: In Table 4, " / " means no such item.

[0079] The cell proliferation rate detection results are shown in Table 5. Compared with the blank control group, the cell proliferation rate of the positive control group was significantly increased by 22.27%, indicating that the model was successfully constructed. The cell activity of sample group 1 was increased by 36.98% compared with the blank control group, and the cell activity of sample group 2 was increased by 36.18% compared with the blank control group, indicating that the red ginseng fermentation liquor in the present application has the effect of improving immunity. In Table 5, the significance of the positive control group compared with the blank control group is represented by #, and p<0.01 is represented by ##; the significance of the sample group compared with the blank control group is represented by *, and p<0.01 is represented by **.

[0080] The cell phagocytosis index detection results are shown in Table 6. The phagocytosis index of sample group 3 was significantly increased by 55.20% compared with the blank control group, and the phagocytosis index of sample group 4 was significantly increased by 32.58% compared with the blank control group. In Table 6, the significance of the sample group compared with the blank control group is represented by *, and p<0.01 is represented by **.

[0081] The above results show that the red ginseng fermentation liquor of the present application has the effect of improving immunity, and the red ginseng fermentation liquor of the present application is added with a sweetener xylitol to prepare a product for improving immunity.

[0082] Table 5 Cell proliferation activity detection results

[0083]

[0084] Table 6 Cell phagocytosis index test results

[0085]

[0086] 2. Simulate in vitro digestion.

[0087] The red ginseng fermentation broth prepared in Example 1 was sequentially reacted in artificial saliva, artificial gastric juice and artificial intestinal juice to simulate the digestion process, and the method was as follows:

[0088] a. Take 500 μL of red ginseng fermentation broth, add 500 μL of artificial saliva in a 37°C water bath, mix well, and place in a 37°C water bath for 5 min.

[0089] b. Add 1000 μL of artificial gastric juice in a 37°C water bath to the red ginseng fermentation broth after artificial saliva digestion, mix well, and place in a 37°C water bath for 2 h.

[0090] c. Continue to add 2000 μL of artificial intestinal juice in a 37°C water bath, mix well, and place in a 37°C water bath for 1 h.

[0091] d. After the above step is completed, place in a 90°C water bath for 5 min.

[0092] e. Centrifuge at 8000 rpm for 10 min, collect the supernatant, and perform detection.

[0093] From the above results, it can be seen that the bacterial outer vesicles in the red ginseng fermentation broth still retain the complete vesicle structure after being sequentially digested by artificial saliva, artificial gastric juice and artificial intestinal juice, indicating that they can resist gastric and intestinal juice digestion, and the bacterial outer vesicles in the red ginseng fermentation broth are stable and have good ability to retain their contents. Figure 4

[0094] 3. Storage stability.

[0095] The red ginseng fermentation broth prepared in Example 1 was added with 0.1% potassium sorbate preservative, and then stored at 5°C and 25°C for a certain period of time, and the pH stability, soluble solids content, color and appearance morphology and bacterial outer vesicle structure indicators were observed and detected, respectively, and the detailed results are shown in Table 7 and Figure 5 .

[0096] The results show that the red ginseng fermentation broth prepared in Example 1 was stored at 5°C and 25°C for 2 months, and the solution was still clear.

[0097] Table 7 Storage stability results

[0098]

[0099] ​In Table 7, "-" indicates that the bacterial outer vesicle structure is not detected. Since the detection time interval is short, the structure of the bacterial outer vesicle does not change much in a short period of time, so it is not detected.

[0100] The technical features of the above-described embodiments can be combined in any manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments are described, but it should be understood that any combination of the technical features is within the scope of the present disclosure as long as the combination does not result in a contradiction.

[0101] The above-described embodiments only express several implementation manners of the present application, and the description is relatively specific and detailed, but it should not be understood as a limitation on the scope of the patent. It should be pointed out that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application, and these are within the protection scope of the present application.

Claims

1. A method for preparing red ginseng fermentation liquid, characterized in that: The following steps are involved: The red ginseng powder is dissolved in water, and then cellulase and pectinase are added to destroy the cell walls of the red ginseng, followed by enzyme inactivation and sterilization to obtain a red ginseng fermentation medium; preparing a seed solution of Lactobacillus plantarum; the Lactobacillus plantarum is YZ-Lactobacillus plantarum LP28; preparing a seed liquid of fermented Lactobacillus mucilaginosus; the fermented Lactobacillus mucilaginosus is fermented Lactobacillus mucilaginosus JYLF-71; mixing the seed liquid of Lactobacillus plantarum and the seed liquid of Lactobacillus mucilaginosus fermentans to obtain a composite strain seed liquid; inoculating the composite strain seed liquid into a red ginseng fermentation medium for fermentation and culturing to obtain a fermentation liquid; The fermentation liquid is centrifuged to collect the supernatant, and the supernatant is squeezed and filtered to collect the filtrate, and the obtained filtrate is the red ginseng fermentation liquid; the pore size of the filter membrane used in the squeeze filtration is 0.8 μm.

2. The preparation method according to claim 1, wherein The volume ratio of the seed liquid of the plant lactobacillus to the seed liquid of the fermentative Lactobacillus mucilaginosus is 1:1, and the inoculation amount of the composite strain seed liquid is 4% by volume.

3. The preparation method according to claim 1, wherein The number of viable Lactobacillus plantarum bacteria in the seed liquid of the Lactobacillus plantarum is 1×10 8 CFU / mL; The number of viable bacteria of fermented Lactobacillus mucilaginosus in the seed liquid of fermented Lactobacillus mucilaginosus is 1×10 8 CFU / mL.

4. The preparation method according to claim 1, wherein The addition amount of the cellulase and pectinase is: Add 0.05 mL of cellulase and 0.05 mL of pectinase to every 1 g of red ginseng powder.

5. The preparation method according to claim 1, wherein The fermentation culture condition is static culture at 37° C. for 48 hours.

6. A red ginseng fermented liquid obtained according to the preparation method according to claim 1.

7. The use of the red ginseng fermentation liquid according to claim 6, characterized in that: The red ginseng fermented liquid is used to prepare products that enhance immunity.

8. The use according to claim 7, characterized in that The product further comprises a pharmaceutically acceptable excipient, which is a sweetener.

9. The use according to claim 8, characterized in that The sweetener is xylitol.

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