Postbiotic composition micro-capsule, preparation method thereof and application of micro-capsule in hangover-alleviating and liver-protecting products

Abstract

The invention provides a retrogen composition microcapsule, a preparation method thereof and application of the retrogen composition microcapsule in hangover alleviating and liver protecting products, and relates to the technical field of retrogen preparation. The postbiotic composition is prepared by the following steps: fermenting lactobacillus mucus Postbio-Q7, lactobacillus paracasei Postbio-P6 and lactobacillus paracasei Postbio-P8, and then inactivating the fermented lactobacillus mucus Postbio-Q7, the fermented lactobacillus paracasei Postbio-P6 and the fermented lactobacillus paracasei Postbio-P8. The preparation method of the postbiotic composition micro-capsule comprises the following steps: 1, adding sodium alginate, shellac, a guarana extract and protamine into sterile deionized water to prepare a micro-capsule shell; and 2, mixing a micro-capsule shell with the metagen composition to prepare the metagen composition micro-capsule. The metagen composition disclosed by the invention has excellent effects of dispelling the effects of alcohol and protecting the liver, and the prepared micro-capsules can play a better role in protecting the metagen composition, so that the metagen composition is convenient to store and transport, internal components are protected, the oxidation value is reduced, and the bacteriostatic and antibacterial effects of the metagen are further improved.

Description

Technical Field

[0001] The present invention relates to the technical field of postbiotic preparation, and in particular to a postbiotic composition microcapsule and a preparation method thereof, and application thereof in alcohol sobering and liver protecting products. Background Art

[0002] In May 2021, the International Scientific Association of Probiotics and Prebiotics (ISAPP) released a consensus statement on postbiotics, making postbiotics officially a category of concern in the field of global intestinal microecology. The statement defines postbiotics as: "A preparation of inanimate microorganisms and / or inanimate microorganisms and / or their components that are beneficial to the health of the host." According to research results, postbiotics play an important role in maintaining the intestinal barrier, regulating immune function, anti-oxidation, and anti-hypertension. At the same time, due to the "inanimate" characteristics of postbiotics, although the research time is far behind probiotics, it still does not prevent the academic and industrial circles from being fascinated by them. The industry regards postbiotics as a new type of microecological product after probiotics, prebiotics and synbiotics, which gives it a broad application prospect.

[0003] my country's wine culture has a long history and has been passed down to this day. Wine has become an important part of our culture, daily life, and work. In daily life, people often drink excessively for various reasons, which causes serious damage to the liver. When alcohol enters the human body, the alcohol dehydrogenase in the liver breaks down ethanol into acetaldehyde, and acetaldehyde dehydrogenase breaks down acetaldehyde into acetic acid. Subsequently, acetaldehyde is broken down into non-toxic water and carbon dioxide through the tricarboxylic acid cycle and excreted from the body. Long-term excessive drinking increases the burden on the liver, and the ability to decompose ethanol and acetaldehyde decreases. Excessive acetaldehyde produced will inhibit the tricarboxylic acid cycle, causing lipid metabolism to be blocked and damaging liver tissue. Therefore, long-term excessive drinking can easily cause alcoholic liver diseases such as alcoholic liver, cirrhosis, liver fibrosis, and even liver cancer.

[0004] Some probiotics have been shown to have beneficial effects on alcoholic fatty liver. The most effective probiotics currently studied are lactic acid bacteria and bifidobacteria, including Lactobacillus rhamnose and animal bifidobacterium subspecies. Clinical studies have found that Lactobacillus rhamnose can enhance the phagocytic function of neutrophils in patients with alcoholic cirrhosis. Animal experimental studies have also confirmed that Lactobacillus rhamnose can improve alcoholic liver damage by reducing liver inflammatory response and maintaining intestinal mucosal integrity. The Chinese patent with publication number CN115895950A discloses the use of Lactobacillus paracasei SMNLBK for hangover or addition to alcoholic fermented beverages. In this comparative document, the probiotics used are live bacteria, not postbiotics prepared by fermented bacteria. The Chinese patent with publication number CN117487690A discloses a method of using at least one of the Lactobacillus rhamnosus MC-1 strain, the Lactobacillus rhamnosus SSN-13 strain and the Limosilactobacillus fermentum G-1-46 strain to prepare a probiotic fermentation agent for fermenting a Chinese medicine composition to prepare a drink with the effect of sobering up and protecting the liver. In this comparative document, on the one hand, active probiotics are used, and on the other hand, the active probiotics are used for fermentation, rather than probiotic postbiotics directly used for sobering up and protecting the liver.

[0005] It should be noted that the information disclosed in the above background technology section is only used to enhance the understanding of the background of the present disclosure, and therefore may include information that does not constitute the prior art known to ordinary technicians in the field. Summary of the invention

[0006] In view of this, the present invention proposes a postbiotic composition microcapsule and a preparation method thereof, and application thereof in alcohol sobering and liver protecting products. A postbiotic composition with liver protecting and alcohol sobering functions is prepared by fermenting three probiotics, namely, Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8, in stages.

[0007] The technical solution of the present invention is achieved as follows: The present invention provides a postbiotic composition, which is obtained by fermenting and inactivating Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8.

[0008] The present invention also provides a postbiotic composition microcapsule, comprising a microcapsule shell and a microcapsule core, wherein the microcapsule core is the postbiotic composition according to claim 1. It is understood that the microcapsule shell can be a common raw material for preparing pharmaceutical microcapsules. The postbiotic composition is wrapped in a microcapsule, which can protect the components in the postbiotic composition and improve the stability of the postbiotic composition in the microcapsule core.

[0009] The present invention also provides a method for preparing the above-mentioned postbiotic composition microcapsules, comprising the following steps:

[0010] S10, adding sodium alginate, shellac, guarana extract and protamine into sterile deionized water, stirring continuously at 50-60° C. for 30-60 min to obtain a dissolved solution;

[0011] Wherein, based on the total mass of 100%, sodium alginate 1% to 4%, shellac 0.5% to 2%, guarana extract 0.1% to 1%, protamine 0.1% to 1%, and the balance is sterile deionized water;

[0012] S20, stirring and homogenizing the dissolved solution at 10000-15000 rpm for 10-15 minutes to obtain a uniform solution, and filtering with a 0.1 μm-0.4 μm filter to obtain a microcapsule shell solution;

[0013] S30, mixing the microcapsule shell solution with the postbiotic composition, homogenizing, and drying; the mass ratio of the microcapsule solution to the postbiotic composition is (1-4): (2-6), the homogenization pressure is 20-40 MPa, and the drying temperature is 160-190° C. to obtain postbiotic composition microcapsules.

[0014] The preparation method uses sodium alginate, shellac, guarana extract and protamine to prepare the shell of the microcapsule to improve the stability and antioxidant capacity of the postbiotic composition. Among them, sodium alginate is a natural polysaccharide with good water solubility and solubility. It mainly plays a role in stabilizing the microcapsule shell structure and emulsification in the microcapsule structure. Shellac is mainly used as a film-forming material in the microcapsule shell, providing good film-forming property and stability, which can extend the shelf life of the raw materials and protect the internal components (postbiotic composition), reduce the oxidation value, and enhance the moisture resistance of the postbiotic composition microcapsules. Protamine in the microcapsule shell mainly improves the antibacterial property and stability of the postbiotic composition microcapsules and extends the shelf life of the postbiotic composition microcapsules. The film-forming material shellac forms a coacervate with sodium alginate and protamine with a cross-linked structure, and then the biochemical substances of the guarana extract interact with the polymer constituting the coacervate to strengthen the microcapsule shell structure. The above four components cooperate with each other and work synergistically to form more stable postbiotic composition microcapsules with the postbiotic composition. Compared with the postbiotic composition microcapsules prepared by using part of the above four components or the postbiotic composition without microcapsules, the solubility of the postbiotic composition microcapsules can be significantly increased, the oxidation value thereof can be reduced, and the fluidity and moisture resistance of the postbiotic composition microcapsules can be increased.

[0015] On the basis of the above technical scheme, preferably, in step S10, based on the total mass of 100%, sodium alginate is 2%, shellac is 0.5%, guarana extract is 0.5%, protamine is 0.2%, and the balance is sterile deionized water. Preferably, in step S20, a 0.2 μm filter is used for filtration.

[0016] The present invention also provides an application of the postbiotic composition microcapsule in preparing a product for sobering up and protecting the liver. For example, the postbiotic composition microcapsule is combined with a pharmaceutically acceptable excipient to prepare a drug for sobering up and protecting the liver.

[0017] The present invention also provides a method for preparing the above-mentioned postbiotic composition, comprising the following steps:

[0018] S1, activating fermented Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 to obtain fermented Lactobacillus mucilaginosus Postbio-Q7 activated bacterial liquid, the number of viable bacteria is (5-10)×10 6 cfu / mL; Lactobacillus paracasei Postbio-P6 activated bacterial solution, the number of viable bacteria is (5~10)×10 6 cfu / mL; and Lactobacillus paracasei Postbio-P8 activated bacterial solution, the number of viable bacteria is (5-10)×10 6 cfu / mL;

[0019] S2, inoculating the activated bacterial solution of Lactobacillus mucilaginosus Postbio-Q7 into the fermentation medium at an inoculation rate of 2% to 4% v / v, performing the first fermentation, and obtaining the first fermentation solution;

[0020] S3, inoculating the activated bacterial solution of Lactobacillus paracasei Postbio-P6 and the activated bacterial solution of Lactobacillus paracasei Postbio-P8 into the first fermentation liquid at an inoculation amount of 2% to 4% v / v, performing a second fermentation to obtain a second fermentation liquid;

[0021] S4. Performing an inactivation treatment on the second fermentation broth to obtain a postbiotic composition.

[0022] After fermenting Lactobacillus mucilaginosus Postbio-Q7 for the first time, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 are added for the second fermentation. In this way, the conditions of the first fermentation and the second fermentation can be adjusted respectively, thereby improving the antibacterial effect of the prepared postbiotic composition.

[0023] It should be noted that the inactivation treatment in step S4 is: the second fermentation broth can be pasteurized at 95°C for 40-100 minutes to obtain a postbiotic fermentation broth, and then the postbiotic fermentation broth is clarified, impurity-removed, concentrated and desalted to obtain a postbiotic composition. Furthermore, the obtained postbiotic composition is in a liquid state. For the convenience of storage and transportation, the postbiotic composition can be freeze-dried to obtain a postbiotic freeze-dried powder.

[0024] On the basis of the above technical scheme, preferably, step S3 comprises: adding a feed growth factor to the first fermentation broth; inoculating the activated bacterial solution of Lactobacillus paracasei Postbio-P6 and the activated bacterial solution of Lactobacillus paracasei Postbio-P8 into the first fermentation broth added with the feed growth factor at an inoculation amount of 2% to 4% v / v, performing a second fermentation to obtain a second fermentation broth;

[0025] The volume of the feed growth factor is 0.04% to 0.06% of the volume of the first fermentation broth. Preferably, the volume of the feed growth factor is 0.05% of the volume of the first fermentation broth.

[0026] Thus, after the first fermentation, before the second addition of probiotics (Lactobacillus paracasei Postbio-P6 activated bacterial solution and Lactobacillus paracasei Postbio-P8), supplementary addition of feed growth factors can improve the efficiency of the second fermentation culture, thereby improving the alcohol sobering and liver protecting effect of the prepared postbiotic composition.

[0027] On the basis of the above technical solution, preferably, in step S2, the fermentation conditions of the first fermentation are: fermentation at 37° C. for 20 to 30 h, a rotation speed of 120 to 180 rpm / min, and a fermentation pH of 4.0 to 4.5;

[0028] In step S3, the fermentation conditions of the second fermentation are: fermentation at 37° C. for 10 to 20 hours, a rotation speed of 120 to 180 rpm / min, and a fermentation pH of 3.2 to 3.8.

[0029] On the basis of the above technical scheme, preferably, in step S3, the feed growth factor includes, by weight, 0.5 parts of Tween 80, 1 part of disodium hydrogen phosphate, 0.01 parts of manganese sulfate, 0.1 parts of magnesium sulfate, 10 parts of yeast powder, 0.02 parts of folic acid, 0.01 parts of biotin, and 0.01 parts of vitamin K.

[0030] The present invention also provides an application of the above-mentioned postbiotic composition in preparing a product for sobering up and protecting the liver. For example, the postbiotic composition is combined with a pharmaceutically acceptable excipient to prepare a product for sobering up and protecting the liver.

[0031] Compared with the prior art, the postbiotic composition and postbiotic composition microcapsule provided by the present disclosure have the following advantages:

[0032] Beneficial effects:

[0033] 1. The postbiotic composition provided by the present invention uses fermented Lactobacillus mucilaginosus Postbio-Q7 for the first stage of fermentation, and then adds Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 for the second stage of fermentation. The fermentation conditions of each fermentation stage can be adjusted respectively, so that the obtained postbiotic composition has excellent alcohol sobering and liver protecting effects.

[0034] 2. By using microcapsule technology to prepare microcapsule-coated postbiotic compositions, the stability, antioxidant properties, moisture resistance and fluidity of the microcapsule shell structure can be improved by combining and synergistically optimizing the raw materials for preparing the microcapsule shell. The stable microcapsule shell can better protect the postbiotic composition, facilitate the storage and transportation of the postbiotic composition, protect the internal components (postbiotic composition), reduce the oxidation value, and thus improve the antibacterial and antimicrobial effects of the postbiotics. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0036] Figure 1 This is a schematic diagram comparing the survival rate results of various groups of human liver cancer cells in one embodiment of the present disclosure.

[0037] Figure 2 This is a schematic diagram of a half inhibition rate curve of human liver cancer cells in one embodiment of the present disclosure.

[0038] Figure 3 This is a schematic diagram of the FDA fluorescence signal intensity of each group of human liver cancer cells in one embodiment of the present disclosure.

[0039] Figure 4 This is a schematic diagram of zebrafish behavior after different treatments are used in the zebrafish drunkenness model in one embodiment of the present invention.

[0040] Figure 5 This is a schematic diagram of the comparison results of the movement distance of zebrafish after different treatments in the zebrafish drunkenness model in one embodiment of the present invention.

[0041] Figure 6 This is a schematic diagram of the fat staining intensity of zebrafish liver after each group of samples were processed in one embodiment of the present disclosure.

[0042] Figure 7 This is a schematic diagram comparing the fat staining intensity (pixels) of each group of zebrafish in one embodiment of the present disclosure.

[0043] Figure 8 This is a schematic diagram of the comparison results of alcohol dehydrogenase content in serum samples of each group in one embodiment of the present disclosure.

[0044] Fig. 9 This is a schematic diagram of the comparison results of acetaldehyde dehydrogenase content in serum samples of each group in one embodiment of the present disclosure.

[0045] Fig.10 This is a schematic diagram of the principal component analysis results of transcription sequencing of each group of samples in one embodiment of the present disclosure.

[0046] Fig.11 This is a volcano plot of differential gene screening between the model control group and the epizootic group in one embodiment of the present disclosure.

[0047] Fig.12 This is a volcano plot of differential gene screening between the model control group and the positive control group in one embodiment of the present disclosure.

[0048] Fig.13 This is a schematic diagram of the KEGG pathway enrichment results of up-regulated differentially expressed genes in the model group and the postbiotic group in one embodiment of the present disclosure.

[0049] Fig.14This is a schematic diagram of the KEGG pathway enrichment results of down-regulated differentially expressed genes in the model group and the postbiotic group in one embodiment of the present disclosure.

[0050] Fig.15 In one embodiment of the present disclosure, postbiotics are used for protection against acute liver injury, and HE staining results of liver tissue sections of mice are compared (×400).

[0051] Fig.16 In one embodiment of the present disclosure, postbiotics are used to repair alcoholic liver damage, and the HE staining results of mouse liver tissue sections are compared (×400). DETAILED DESCRIPTION

[0052] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0053] The fermented mucus Lactobacillus (Limosilactobacillus fermentum) Postbio-Q7, Lactobacillus paracasei (Lactobacillus paracasei) Postbio-P6 and Lactobacillus paracasei (Lacticaseibacillus paracasei) Postbio-P8 used in the present invention are all preserved in the General Microbiological Center of China Microorganism Culture Collection Administration, and the preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The preservation numbers are CGMCC No. 26238, CGMCC No. 26237 and CGMCC No. 31112, respectively, and the preservation dates are December 25, 2022, December 25, 2022 and June 27, 2024, respectively.

[0054] The above-mentioned postbiotic composition and postbiotic composition microcapsule are further described below in conjunction with specific examples.

[0055] The basic formula of the fermentation medium used in the following examples is: 20 g glucose, 15 g whey protein, 20 g soy flour, 10 g skim milk powder, 10 g yeast powder, 1 L distilled water, initial pH = 7.0. Before inoculation and fermentation, the fermentation medium needs to be sterilized at 105°C for 30 min.

[0056] The formula of the supplement growth factor is: Tween 80 0.5g, disodium hydrogen phosphate 1g, manganese sulfate 0.01g, magnesium sulfate 0.1g, yeast powder 10g, folic acid 0.02g, biotin 0.01g, vitamin K 0.01g.

[0057] Example 1

[0058] S1. Inoculate fermented Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 into MRS liquid culture medium, culture at 37°C for 18 h, and subculture twice to make the number of viable bacteria of each strain after activation be (5-10)×10 6 cfu / mL.

[0059] S2. Inoculate the activated bacterial solution of Lactobacillus mucilaginosus Postbio-Q7 into the fermentation medium at an inoculation rate of 2% v / v, and ferment at 37° C. for 25 h, with a rotation speed of 150 rpm / min and a fermentation pH of 4.0-4.5 to obtain a first fermentation solution.

[0060] S3, adding 0.05% v / v of feed growth factor to the first fermentation broth, inoculating the first fermentation broth added with the feed growth factor at an inoculation rate of 2% v / v with activated bacterial solution of Lactobacillus paracasei Postbio-P6 and activated bacterial solution of Lactobacillus paracasei Postbio-P8, and performing a second fermentation to obtain a second fermentation broth.

[0061] S4. After the fermentation, the second fermentation liquid is pasteurized at 95° C. for 40-100 minutes to obtain a postbiotic fermentation liquid. The postbiotic fermentation liquid is then clarified, impurity-removed, concentrated and desalted to obtain a postbiotic composition solution, which is marked as postbiotic composition 1.

[0062] Example 2

[0063] The difference between this embodiment and embodiment 1 is that no feed growth factor is added in step S3, and the remaining steps are the same as those in embodiment 1. The obtained postbiotic composition solution is marked as postbiotic composition 2.

[0064] Example 3

[0065] The difference between this embodiment and embodiment 1 is that fermented mucus lactobacillus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 are inoculated in a fermentation medium at a rate of 2% v / v, respectively, and fermented at 37°C for 25 hours, with a rotation speed of 150rpm / min and a fermentation pH of 4.0-4.5. After one fermentation, sterilization, clarification and impurity removal and concentration and desalination treatment in step S4 are directly performed. That is to say, step S2 and step S3 are combined, and only one fermentation is performed. The remaining steps are the same as in embodiment 1, and the obtained postbiotic composition solution is marked as postbiotic composition 3.

[0066] The moisture content, solubility, oxidative stability, fluidity and moisture resistance of the postbiotic compositions prepared in Examples 1 to 3 were tested.

[0067] Solubility measurement method: Accurately weigh 1 g of the postbiotic composition sample prepared in the above embodiment, dissolve it in 25 mL of deionized water, transfer all the sample solution into a centrifuge tube and centrifuge it at 3000×g for 10 min. After decanting the supernatant, add a small amount of deionized water, centrifuge again to separate the supernatant, transfer the precipitate into a watch dish that has been dried to constant weight, and dry it in an oven at 105°C to constant weight. The mass of the sample minus the mass of the precipitate is the solubility of the sample.

[0068] Moisture resistance test method: Weigh 5 g of the postbiotic composition sample prepared in the above embodiment, place it at 20°C and 75% humidity for 12 hours, measure its weight change, calculate the weight gain rate, and calculate the moisture resistance of the corresponding sample according to the change in weight gain rate.

[0069] In this example, the moisture content of the postbiotic composition sample was determined by the oven method according to GB / T5528-2008. The oxidative stability of the postbiotic composition sample was tested according to the colorimetric method in GB / T 5009.37-2003. The fluidity of the postbiotic composition sample was tested by the shrinkage (CI) and the Hausner ratio (HR) reaction. The results are shown in Table 1.

[0070] Among them, for fluidity, generally speaking, CI<10, HR between 1.00 and 1.11 indicates that the powder fluidity is very good; CI<15, HR between 1.12 and 1.18 indicates that the powder fluidity is good; CI>16, HR>1.2 indicates that the powder fluidity is average or poor. The meq / kg of oxidation stability means milliequivalent per kilogram. meq is the abbreviation of milliequivalent, a unit of measurement for the chemical activity or reactivity of a substance.

[0071] Table 1. Comparison of performance of postbiotic compositions 1-3

[0072]

[0073] It can be seen from the results in Table 1 that, compared with postbiotic composition 2 and postbiotic composition 3, the postbiotic composition obtained by culturing Lactobacillus mucilaginosus Postbio-Q7 with Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 in stages has low water content, improved solubility, strong oxidative stability and improved moisture resistance.

[0074] The postbiotic composition prepared in Example 1 is used below to verify the alcohol sobering and liver protecting effects of the postbiotic composition.

[0075] 1. Cytotoxicity testing of postbiotic compositions

[0076] The postbiotic composition obtained in Example 1 is centrifuged at 4000rpm / min for 10min, the supernatant is collected, and the supernatant is filtered through a 0.22μm filter membrane to obtain postbiotics. The postbiotics are configured into postbiotic solutions with contents of 3.125%, 6.25%, 12.5%, 25%, 50%, and 100%, respectively. For example, a 50% postbiotic solution is obtained by diluting the postbiotics by half with a culture medium; a 100% postbiotic solution is a postbiotic directly filtered with a 0.22μm filter membrane.

[0077] Human hepatoma cells were expressed as 5×10 4 Cells were inoculated into a 96-well plate at a seeding density of 10 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ) until the cell plating rate in the 96-well plate reached 40% to 60%, and then postbiotics were added for the experiment.

[0078] Experimental group: divided into 6 groups according to the different concentrations of the added postbiotic solution, and the postbiotic solution added to each group was 3.125%, 6.25%, 12.5%, 25%, 50%, and 100%, respectively; 200 μL of the postbiotic solution of the corresponding solubility was added to each well.

[0079] Blank control group: 200 μL of culture medium was added to each well.

[0080] Positive control group: 200 μL of culture medium containing 10% DMSO was added to each well.

[0081] After the administration, the 96-well plate was placed in an incubator (37°C, 5% CO2) for 24 hours. After the cells were incubated for 24 hours, the supernatant was discarded, and MTT working solution (0.5 mg / mL) was added, and the cells were incubated at 37°C in the dark for 4 hours. After the incubation, the supernatant was discarded, 150 μL of DMSO was added to each well, and the cells were shaken for 15 minutes, and the OD value was read at 490 nm.

[0082] like Figure 1As shown, when postbiotics were added to human liver cancer cells for co-culture, the cell survival rate reached more than 90% when the postbiotics were added at a concentration range of 6.25%. Compared with the blank control group, no obvious cytotoxicity to liver cancer cells was shown. When the postbiotics were added at a concentration of 12.5% ​​and above, the cell survival rate showed a significant difference compared with the blank control group, and the cell survival rate decreased in a concentration-dependent manner. However, when 100% postbiotics were added, the cell survival rate was still higher than that of the positive control group. This experiment shows that postbiotics did not cause obvious damage to cells within a concentration range of 6.25%, but when the solubility increased to 12.5%, the postbiotics began to show cytotoxicity to liver cancer cells. As Figure 2 As shown, according to the cell half inhibition rate curve, when 50% of human liver cancer cells are apoptotic, the concentration of postbiotics is 33.6%.

[0083] 2. Effects of postbiotic composition on cell membrane permeability

[0084] Fluorescein diacetate (FDA) staining method is used to evaluate cell viability and survival. The basic principle is that FDA is a fat-soluble substance that is non-fluorescent. It can freely pass through the cell membrane into living cells and can be decomposed by intracellular esterase to produce a green fluorescent substance, fluorescein, which accumulates in the cells.

[0085] The postbiotic composition obtained in Example 1 was centrifuged at 4000 rpm / min for 10 min, the supernatant was collected, and the supernatant was filtered through a 0.22 μm filter membrane to obtain postbiotics. The postbiotics were configured into postbiotic solutions with contents of 3.125%, 6.25%, 12.5%, 25%, 50%, and 100%, respectively.

[0086] Human hepatoma cells were expressed as 5×10 4 Cells were inoculated into a 96-well plate at a seeding density of 10 cells / well and incubated overnight in an incubator (37° C., 5% CO 2 ) until the cell plating rate in the 96-well plate reached 40% to 60%, and then postbiotics were added for the experiment.

[0087] Experimental group: divided into 6 groups according to the different concentrations of the added postbiotic solution, and the postbiotic solution added to each group was 3.125%, 6.25%, 12.5%, 25%, 50%, and 100%, respectively; 200 μL of the postbiotic solution of the corresponding solubility was added to each well.

[0088] Control group (0% Postbiotic): 200 μL of culture medium was added to each well.

[0089] After the administration is completed, the 96-well plate is placed in an incubator (37°C, 5% CO2) for 24 hours. Take out the cells to be tested and wash them twice with PBS to remove the interfering components in the culture medium and postbiotics. Add FDA solution separately and incubate human liver cancer cells with FDA at 37°C for 15 minutes to ensure that the FDA solution covers the cell surface. After 15 minutes, remove the FDA solution and wash the cells twice with PBS to remove the FDA solution that has not entered the cells. Observe the cells under a fluorescence microscope and record the fluorescence signal. Live cells should show green fluorescence signals, and dead cells should not show fluorescence.

[0090] Results Figure 3 After adding 12.5% ​​postbiotic solution (12.5% ​​Postbiotic in the figure), the viable cell rate was more than 90% compared with the normal group without postbiotic (0% Postbiotic), indicating that when the concentration of postbiotic solution was 12.5%, the cells did not show obvious cytotoxicity. With the increase of the concentration of postbiotic solution, the green fluorescence signal of 25% postbiotic solution (25% Postbiotic in the figure) and 50% postbiotic solution (50% Postbiotic in the figure) was significantly weakened, and the cytotoxicity of postbiotic solution was significantly enhanced, indicating that postbiotic solution can act on human liver cancer cells and inhibit the activity of human liver cancer cells.

[0091] 3. The hangover-relief effect of postbiotic combination

[0092] The postbiotic composition obtained in Example 1 was centrifuged at 4000 rpm / min for 10 min, the supernatant was collected, and the supernatant was filtered through a 0.22 μm filter membrane to obtain postbiotics. The postbiotics were prepared with water to form postbiotic solutions with contents of 0.78 μl / ml, 1.56 μL / mL, and 3.12 μl / ml, respectively.

[0093] Construction of drunken animal model: Genetically mutant Albino zebrafish at 5 days post-fertilization (dpf) were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well. After each group was induced with anhydrous ethanol for 1 hour, the ethanol was removed to establish the zebrafish drunken model.

[0094] Experimental group: including three groups ( Figure 4 The indicated postbiotics were postbiocare 0.78 μl / ml, postbiocare 1.56 μl / ml, and postbiocare 3.12 μl / ml). 200 μl of postbiotic solution with a concentration of 0.78 μl / ml, 1.56 μL / mL, and 3.12 μl / ml was added to each group, respectively, and treated with the zebrafish drunkenness model at 28°C for 1 hour.

[0095] Model control group: 200 μl of water was added to the wells with zebrafish drunkenness model and treated at 28°C for 1 h.

[0096] Positive control group (RU21 100 μg / mL): 200 μl of RU21 (RU-21 Antipol natural plant extract) solution with a concentration of 100 μg / mL was added to the wells with the zebrafish drunkenness model and treated at 28° C. for 1 hour.

[0097] Normal control group: 200 μl of water was added to the wells of normal zebrafish and treated at 28°C for 1 h.

[0098] Ten zebrafish were randomly selected from each group and placed in a 96-well plate, with one zebrafish in each well and a capacity of 200 μL per well. The total movement distance of the zebrafish was detected by a behavior analyzer, and the statistical analysis results of this index were used to evaluate the alcohol sobering effect of the sample. The statistical processing results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0099] The results are as follows Figure 4 As shown, the black line is the slow movement distance, the green line is the medium speed movement distance, and the red line is the fast movement distance. When alcohol was added, the movement behavior of zebrafish in the model group was mainly slow, accompanied by a small amount of medium speed behavior, while the blank control group and the positive control group ( Figure 4 The RU21 100 μg / mL group shown in the figure showed mainly low and medium speed behaviors, accompanied by a small amount of high speed behaviors. When the dosage of postbiotics reached 3.12 μl / ml, the movement behavior of zebrafish was not significantly different from that of the blank control group and the positive control group. Figure 5 When the concentration of postbiotics reached 3.12 μL / mL, the total movement distance of drunk zebrafish was basically the same as that of the blank control group and the positive control group, and was significantly different from the model group. This shows that postbiotics have a significant effect of sobering up after drinking at a dose of 3.12 μL / mL. This shows that when a high concentration of postbiotic solution is added, it can play a role in sobering up and normalize the behavior of zebrafish.

[0100] 4. The liver-protecting effect of the postbiotic combination after drinking

[0101] The postbiotic composition obtained in Example 1 was centrifuged at 4000 rpm / min for 10 min, the supernatant was collected, and the supernatant was filtered through a 0.22 μm filter membrane to obtain postbiotics. The postbiotics were prepared with water to form postbiotic solutions with contents of 0.78 μl / ml, 1.56 μL / mL, and 3.12 μl / ml, respectively.

[0102] Albino mutant zebrafish at 5 days post fertilization (dpf) were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well.

[0103] Experimental group: There were three groups, each of which was added with 200 μl of postbiotic solution with concentrations of 0.78 μl / ml, 1.56 μL / mL, and 3.12 μl / ml, respectively, and then anhydrous ethanol was added and treated at 28°C for 1 day (1d).

[0104] Model control group: anhydrous ethanol was added to the corresponding wells and treated at 28°C for 1 day (1d).

[0105] Positive control group: RU21 at a concentration of 100 μg / mL was added to the corresponding medium, and then anhydrous ethanol was added and treated at 28°C for 1 day (1d).

[0106] Blank control group: only aqueous solution was added to the corresponding wells.

[0107] After the above treatment, the serum of the corresponding zebrafish in each group was collected for alcohol dehydrogenase and acetaldehyde dehydrogenase content detection. The fish body was fixed and stained with Oil Red O (a fat-soluble dye). After staining, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. The data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The liver fat staining intensity was analyzed, and the statistical analysis results of this indicator were used to evaluate the protective effect of the sample on alcoholic fatty liver. The statistical processing results were expressed as mean ± SE. SPSS26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0108] Results Figure 6-Figure 9 ,according to Figure 6 The experimental results show that after adding alcohol treatment, the liver fat content of the positive control group and the experimental group with a postbiotic concentration of 1.56μl / ml decreased accordingly compared with the model control group. This shows that the degree of alcohol-induced zebrafish fatty liver in the positive control group and the experimental group with a postbiotic concentration of 1.56μl / ml is relatively mild, that is, the addition of RU21 and postbiotic solution can reduce the induction of alcohol on fatty liver, and the postbiotic solution with a concentration of 1.56ml / ml has a protective effect on the liver. Figure 7 As shown, except for the blank control group, the experimental group with a postbiotic concentration of 1.56 ml / ml showed a significant difference compared with the model control group, indicating that the liver protection effect of postbiotics is better when added at 1.56 ml / ml. Figure 8 and Fig. 9 The test results showed that after adding postbiotics, the levels of alcohol dehydrogenase and acetaldehyde dehydrogenase in zebrafish were significantly increased compared with the model group, and the changes in the levels of alcohol dehydrogenase and acetaldehyde dehydrogenase were most significant in the experimental group with a postbiotic concentration of 1.56ml / ml, proving that when the concentration of the postbiotic solution is 1.56ml / ml, the liver protection effect is higher, and the postbiotic solution has an optimal concentration range for liver protection.

[0109] 5. Effects of postbiotic composition on gene transcription expression in zebrafish

[0110] The postbiotic composition obtained in Example 1 was centrifuged at 4000 rpm / min for 10 min, the supernatant was collected, and the supernatant was filtered through a 0.22 μm filter membrane to obtain postbiotics. The postbiotics were prepared with water to form postbiotic solutions with contents of 0.78 μl / ml, 1.56 μL / mL, and 3.12 μl / ml, respectively.

[0111] Albino mutant zebrafish at 5 days post fertilization (dpf) were randomly selected and placed in a 6-well plate, with 30 zebrafish in each well.

[0112] Experimental group (postbiotic group): Add 200 μl of postbiotic solution with a concentration of 1.56 μL / mL, then add anhydrous ethanol, and treat at 28°C for 1 day (1d).

[0113] Model control group: anhydrous ethanol was added to the corresponding wells and treated at 28°C for 1 day (1d).

[0114] Positive control group: RU21 at a concentration of 100 μg / mL was added to the corresponding medium, and then anhydrous ethanol was added and treated at 28°C for 1 day (1d).

[0115] Blank control group: only aqueous solution was added to the corresponding wells.

[0116] Four zebrafish from each group were randomly selected and stored in RNA cryoprecipitate for subsequent RNA extraction and transcription sequencing. Sequencing was performed using the Lumina HiSeq platform, and Bowtie 2 software was used for reference sequence comparison analysis. HTSeq and DESeq 2 software were used to calculate gene expression and differential expression. Fig.10 As shown, the samples in the postbiotic group have little difference, the samples are clustered together, and the experimental data are reliable. Compared with the model control group, the data of the postbiotic group and the model control group are more separated, indicating that there are significant differences between the postbiotic group and the model control group after treatment.

[0117] The screening criteria of Fold Change ≥ 1.5 and P value < 0.05 were used to screen differentially expressed genes. Fig.11 As shown in Figure 2, compared with the model control group, the epibiotic group had a total of 479 differentially expressed genes (DEGs), of which 325 DEGs were upregulated and 154 DEGs were downregulated. Fig.12 As shown, compared with the model control group, there were 3365 DEGs in the positive control group, of which 156 DEGs were upregulated and 209 DEGs were downregulated.

[0118] The significantly up-regulated and down-regulated differentially expressed genes in the metabiotic group were subjected to KEGG enrichment analysis, among which the significantly up-regulated differentially expressed genes were enriched in 88 metabolic pathways, and the significantly down-regulated differentially expressed genes were enriched in 61 metabolic pathways. Fig.13 As shown in the figure, the significantly up-regulated genes were mainly concentrated in the pathways of Primary bile acid biosynthesis, Cytokine-cytokinereceptor interaction, Cell adhesion molecules, Glycosaminoglycan biosynthesis-chondroitin sulfate / dermatan sulfate, Glycosaminoglycanbiosynthesis-keratansulfate, One carbon pool by folate, etc. The up-regulation of these pathways plays an important role in fatty acid metabolism, synthesis, repair and methylation of intracellular DNA, signal transduction, stability of extracellular matrix structure and function, antioxidant capacity of cells and tissue repair. Fig.14 As shown in the figure, significantly down-regulated genes are significantly enriched in pathways such as Endocytosis, Tight junction, MAPK signaling pathway, and Calcium signaling pathway. These pathways have important effects on intracellular signal transduction, calcium ion balance, and responses to various external stimuli.

[0119] Effects of postbiotic composition on acute alcoholic liver injury in mice

[0120] The experimental animals were SPF KM mice. After one week of adaptive feeding, they were randomly divided into groups, 8 mice in each group, and the drug was administered by gavage at 10 mL / kg / mouse.

[0121] Blank group: distilled water was administered by gavage. After 7 days, the mice were gavaged with distilled water at a rate of 14 mL / kg per mouse once. The mice were fasted for 12 hours and then killed.

[0122] Model group: distilled water was administered by intragastric gavage. After 7 days, 50% ethanol solution (analytical grade) was administered once by intragastric gavage at the rate of 14 mL / kg / mouse. The mice were fasted for 12 hours and then killed.

[0123] Positive group: bifendate dripping pills were administered by intragastric gavage. After 7 days, 14 mL / kg / mouse of 50% ethanol solution (analytical grade) was administered once by intragastric gavage. The mice were fasted for 12 hours and then killed.

[0124] Postbiotic low-dose group: 10.835 μl / 10g of postbiotic solution was administered by intragastric gavage. After 7 days, 50% ethanol solution (analytical grade) was administered once by intragastric gavage at a rate of 14 mL / kg / mouse. The mice were fasted for 12 hours and then killed.

[0125] Postbiotic medium-dose group: 21.67 μl / 10g of postbiotic solution was administered by intragastric gavage. After 7 days, 14 mL / kg / mouse of 50% ethanol solution (analytical grade) was administered once by intragastric gavage. The mice were fasted for 12 hours and then killed.

[0126] Postbiotic high-dose group: 43.34 μl / 10g of postbiotic solution was administered by intragastric gavage. After 7 days, 14 mL / kg / mouse of 50% ethanol solution (analytical grade) was administered once by intragastric gavage. The mice were fasted for 12 hours and then killed.

[0127] After the mice in the above groups were killed, the livers were taken out and fixed in paraformaldehyde for 4 days. The fixed mouse livers were taken out, and the parts with intact structures were selected to be removed with a thickness of about 0.5 cm, placed under running tap water for about 24 hours, and dehydrated with gradient alcohol in a 45°C constant temperature box. After completion, xylene was used for transparency, and paraffin sections were made. After the production was completed, they were stained and observed according to the HE staining method.

[0128] The results are as follows Fig.15 As shown in the figure, the liver tissue cells of the blank group mice were arranged regularly and neatly, the intercellular gaps were uniform, the cytoplasm was rich, and no symptoms appeared. The liver cells of the model group mice were irregular in morphology, and a large number of hepatocytes were observed to have vacuolar degeneration, cell swelling, loose and lightly stained cytoplasm around the central vein, portal area and parenchyma. The surrounding hepatic sinusoids were squeezed, and obvious vascular congestion was observed, with obvious pathological changes. The hepatocytes in the central vein and portal area of ​​the postbiotic medium and high dose groups were arranged regularly and orderly, with uniform intercellular gaps, and occasionally mild vacuoles appeared in the hepatocytes. There was no obvious congestion in the blood vessels of the low dose group, and the congestion in the medium and high dose groups was also significantly reduced. Compared with the model group, the symptoms were significantly alleviated. This shows that postbiotics have a significant protective effect on acute alcoholic liver injury.

[0129] VII. Repair effect of postbiotic combination on alcoholic liver damage in mice

[0130] The experimental animals were SPF KM mice, which were randomly divided into groups after one week of adaptive feeding, with 8 mice in each group.

[0131] Blank group: mice were gavaged with distilled water at 6 g / kg / mouse, once every 12 hours, for a total of 3 times. Then, mice were gavaged with distilled water at 10 mL / kg / mouse, for 7 days, fasted for 12 hours, and then killed.

[0132] Model group: 40% ethanol (analytical grade) was gavaged at 6 g / kg / mouse, once every 12 hours, for a total of 3 times. Then, distilled water was gavaged at 10 mL / kg / mouse. After 7 days of gavage, the mice were fasted for 12 hours and then killed.

[0133] Positive group: 6 g / kg / mouse 40% ethanol (analytical grade) was gavaged once every 12 hours for a total of 3 times. Then 10 mL / kg / mouse bifendate dripping pills were gavaged for 7 days, and the mice were fasted for 12 hours and then killed.

[0134] Postbiotic low-dose group: 40% ethanol (analytical grade) was gavaged at 6 g / kg / mouse, once every 12 hours, for a total of 3 times. Then 10.835 μL / 10g postbiotic solution was gavaged at 10 mL / kg / mouse. After 7 days of gavage, the mice were fasted for 12 hours and then killed.

[0135] Postbiotic medium dose group: 40% ethanol (analytical grade) was gavaged at 6 g / kg / mouse, once every 12 hours, for a total of 3 times. Then 21.67 μL / 10g postbiotic solution was gavaged at 10 mL / kg / mouse. After 7 days of gavage, the mice were fasted for 12 hours and then killed.

[0136] Postbiotic high-dose group: 40% ethanol (analytical grade) was gavaged at 6 g / kg / mouse, once every 12 hours, for a total of 3 times. Then 43.34 μL / 10g postbiotic solution was gavaged at 10 mL / kg / mouse. After 7 days of gavage, the mice were fasted for 12 hours and then killed.

[0137] After the mice in the above groups were killed, the livers were taken out and fixed in paraformaldehyde for 4 days. The fixed mouse livers were taken out, and the parts with intact structures were selected to be removed with a thickness of about 0.5 cm, placed under running tap water for about 24 hours, and dehydrated with gradient alcohol in a 45°C constant temperature box. After completion, xylene was used for transparency, and paraffin sections were made. After the production was completed, they were stained and observed according to the HE staining method.

[0138] The results are as follows Fig.16 As shown in the figure, the liver tissue cells of the blank group mice were arranged neatly, the intercellular spaces were uniform, the cytoplasm was abundant, the nuclei were obvious, and no obvious symptoms appeared. The liver tissue cells of the model group mice were arranged chaotically, the intercellular spaces were uneven, the cell edges were irregular, a large number of vacuoles were visible in the cells, and occasionally the cells were slightly swollen and the cytoplasm was overflowed. The liver cells in the high-dose group of the postbiotic group were arranged relatively neatly and orderly, the intercellular spaces were uniform, the cell edges were regular and obvious, and mild vacuoles were rarely seen in the liver cells. Compared with the model group, the symptoms were significantly alleviated. This shows that postbiotics have a repairing effect on acute alcoholic liver damage.

[0139] In order to further improve the stability of postbiotics, the postbiotic composition prepared in Example 1 can be encapsulated by microencapsulation technology to obtain postbiotic composition microcapsules with high stability and good antibacterial performance.

[0140] Example 4

[0141] S10. Add 2 g of sodium alginate, 0.5 g of shellac, 0.5 g of guarana extract and 0.2 g of protamine to 96.8 g of sterile deionized water, and stir continuously at 60° C. for 40 min to obtain a dissolved solution.

[0142] S20, stirring and homogenizing the dissolved solution at 12000 rpm for 15 minutes to obtain a uniform solution, and filtering with a 0.2 μm filter to obtain a microcapsule shell solution.

[0143] S30, mixing the microcapsule shell solution with the postbiotic composition 1, homogenizing, and drying; the homogenizing pressure is 30 MPa, and the spray drying temperature is 180° C. to obtain a postbiotic composition microcapsule marked as postbiotic composition microcapsule 1.

[0144] Example 5

[0145] The difference between this embodiment and embodiment 4 is that in step S10, 2 g of sodium alginate, 0.5 g of shellac, and 0.5 g of guarana extract are added to 97 g of sterile deionized water, and stirred for 40 min at 60° C. to obtain a dissolved solution. The other steps are the same as those in embodiment 4, and a postbiotic composition microcapsule is obtained, which is marked as postbiotic composition microcapsule 2.

[0146] Example 6

[0147] The difference between this embodiment and embodiment 4 is that in step S10, 2 g of sodium alginate, 0.5 g of shellac and 0.2 g of protamine are added to 97.3 g of sterile deionized water, and stirred for 40 min at 60° C. to obtain a dissolved solution. The other steps are the same as those in embodiment 4, and a postbiotic composition microcapsule is obtained, which is marked as postbiotic composition microcapsule 3.

[0148] Example 7

[0149] The difference between this embodiment and embodiment 4 is that in step S10, 2 g of sodium alginate and 0.5 g of shellac are added to 97.5 g of sterile deionized water, and stirred for 40 min at 60° C. to obtain a dissolved solution. The other steps are the same as those in embodiment 4, and a postbiotic composition microcapsule is obtained, which is marked as postbiotic composition microcapsule 4.

[0150] Example 8

[0151] The difference between this embodiment and embodiment 4 is that the postbiotic composition 1 is freeze-dried to obtain a postbiotic composition freeze-dried powder, and the postbiotic composition freeze-dried powder is used to compare the performance with the postbiotic composition microcapsules 1-4 prepared in the above embodiments.

[0152] The moisture content, solubility, oxidative stability, fluidity and moisture resistance of the postbiotic composition microcapsules 1-4 and the postbiotic composition freeze-dried powder were tested respectively. The results are shown in Table 2.

[0153] Table 2. Performance comparison of postbiotic composition microcapsules 1-4 and postbiotic composition freeze-dried powder

[0154]

[0155] As shown in Table 2, compared with the freeze-dried powder of the postbiotic composition, the microcapsules (postbiotic composition microcapsules 1) formed by encapsulating the postbiotic composition with microcapsule shells prepared with sodium alginate, shellac, guarana extract and protamine have greatly improved properties such as antioxidant properties, moisture resistance and solubility. This indicates that the microcapsule shell formed by this formula can protect the postbiotic composition, thereby improving the alcohol sobering and liver protecting effects of the postbiotics.

[0156] Compared with postbiotic composition microcapsule 2, postbiotic composition microcapsule 3 and postbiotic composition microcapsule 4, the performance of postbiotic composition microcapsule 1 is better. This is because no protamine is added to the microcapsule shell of postbiotic composition microcapsule 2, which reduces the stability and antibacterial properties of the microcapsule; the microcapsule shells of postbiotic composition microcapsule 3 and postbiotic composition microcapsule 4 also lack some components, which also affects the cross-linking strength and stability of the microcapsule shell, thereby reducing the protective effect of the postbiotic composition.

[0157] To sum up, the postbiotic composition microcapsules formed by encapsulating the postbiotic composition with microcapsule shells prepared by the synergistic combination of sodium alginate, shellac, guarana extract and protamine can better play an antioxidant and moisture-proof effect, thereby achieving the protection of the postbiotic composition, reducing the difficulty of storage and transportation of the postbiotic composition, and providing greater convenience for the application of the postbiotic composition.

[0158] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A postbiotic composition, characterized in that: The postbiotic composition is obtained by fermenting and inactivating Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8.

2. A postbiotic composition microcapsule, characterized in that: It comprises a microcapsule shell and a microcapsule core, and the microcapsule core is the postbiotic composition according to claim 1.

3. A method for preparing the postbiotic composition microcapsules according to claim 2, characterized in that: The following steps are involved: S10, adding sodium alginate, shellac, guarana extract and protamine into sterile deionized water, stirring continuously at 50-60° C. for 30-60 min to obtain a dissolved solution; Wherein, based on the total mass of 100%, sodium alginate 1% to 4%, shellac 0.5% to 2%, guarana extract 0.1% to 1%, protamine 0.1% to 1%, and the balance is sterile deionized water; S20, stirring and homogenizing the dissolved solution at 10000-15000 rpm for 10-15 minutes to obtain a uniform solution, and filtering with a 0.1 μm-0.4 μm filter to obtain a microcapsule shell solution; S30, mixing the microcapsule shell solution with the postbiotic composition, homogenizing, and drying; the mass ratio of the microcapsule solution to the postbiotic composition is (1-4): (2-6), the homogenization pressure is 20-40 MPa, and the drying temperature is 160-190° C. to obtain postbiotic composition microcapsules.

4. The method for preparing the postbiotic composition microcapsules according to claim 3, characterized in that: In step S10, based on the total mass being 100%, the sodium alginate is 2%, the shellac is 0.5%, the guarana extract is 0.5%, the protamine is 0.2%, and the balance is sterile deionized water.

5. Use of the postbiotic composition microcapsules according to claim 2 in the preparation of alcohol sobering and liver protecting products.

6. A method for preparing the postbiotic composition according to claim 1, characterized in that: The following steps are involved: S1, activating fermented Lactobacillus mucilaginosus Postbio-Q7, Lactobacillus paracasei Postbio-P6 and Lactobacillus paracasei Postbio-P8 to obtain fermented Lactobacillus mucilaginosus Postbio-Q7 activated bacterial liquid, the number of viable bacteria is (5-10)×10 6 cfu / mL; Lactobacillus paracasei Postbio-P6 activated bacterial solution, the number of viable bacteria is (5~10)×10 6 cfu / mL; and Lactobacillus paracasei Postbio-P8 activated bacterial solution, the number of viable bacteria is (5-10)×10 6 cfu / mL; S2, inoculating the activated bacterial solution of Lactobacillus mucilaginosus Postbio-Q7 into the fermentation medium at an inoculation rate of 2% to 4% v / v, performing the first fermentation, and obtaining the first fermentation solution; S3, inoculating the activated bacterial solution of Lactobacillus paracasei Postbio-P6 and the activated bacterial solution of Lactobacillus paracasei Postbio-P8 into the first fermentation liquid at an inoculation amount of 2% to 4% v / v, performing a second fermentation to obtain a second fermentation liquid; S4. Performing an inactivation treatment on the second fermentation broth to obtain a postbiotic composition.

7. The preparation method according to claim 6, characterized in that: Step S3 is: adding a feed growth factor to the first fermentation liquid; inoculating the activated bacterial solution of Lactobacillus paracasei Postbio-P6 and the activated bacterial solution of Lactobacillus paracasei Postbio-P8 at an inoculation amount of 2% to 4% v / v into the first fermentation liquid added with the feed growth factor, performing a second fermentation, and obtaining a second fermentation liquid; Wherein, the volume of the feed growth factor is 0.04-0.06% of the volume of the first fermentation liquid.

8. The preparation method according to claim 6 or 7, characterized in that: In step S2, the fermentation conditions of the first fermentation are: fermentation at 37° C. for 20 to 30 hours, a rotation speed of 120 to 180 rpm / min, and a fermentation pH of 4.0 to 4.5; In step S3, the fermentation conditions of the second fermentation are: fermentation at 37° C. for 10 to 20 hours, a rotation speed of 120 to 180 rpm / min, and a fermentation pH of 3.2 to 3.

8.

9. The preparation method according to any one of claim 7, characterized in that: In step S3, the feed growth factor includes, by mass fraction, 0.5 parts of Tween 80, 1 part of disodium hydrogen phosphate, 0.01 parts of manganese sulfate, 0.1 parts of magnesium sulfate, 10 parts of yeast powder, 0.02 parts of folic acid, 0.01 parts of biotin, and 0.01 parts of vitamin K.

10. Use of the postbiotic composition according to claim 1 in preparing a hangover and liver protection product.

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