Bifidobacterium longum HSBL001 metagen, compound metagen preparation process and application thereof

Abstract

The invention discloses a preparation process and application of a bifidobacterium longum HSBL001 metagen and a compounded metagen, and relates to the technical field of microorganisms. The invention provides a bifidobacterium longum HSBL001, and the preservation number of the bifidobacterium longum HSBL001 is CGMCC (China General Microbiological Culture Collection Center) No.31244. The invention also provides a preparation method of the bifidobacterium longum HSBL001. The invention also provides a fermentation medium of the bifidobacterium longum, and the preparation method comprises the following steps: dissolving the skimmed milk powder and the soybean powder or the skimmed milk powder and the soybean protein isolate, adding the papain for enzymolysis, and adding the pH regulator after enzyme deactivation and sterilization to obtain the fermentation medium. The culture medium can increase the fermentation viable count of the bifidobacterium longum, and the bifidobacterium longum metagen and the bifidobacterium longum compound metagen prepared by using the culture medium have the effect of relieving colitis.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and in particular to a Bifidobacterium longum HSBL001 postbiotic, a preparation process of a composite postbiotic and an application thereof. Background Art

[0002] Bifidobacterium is a non-motile, non-spore-producing, non-gas-producing, Gram-negative, anaerobic bacteria with high GC content. Its morphology is often divergent or irregular "V" or "Y" shaped rod-shaped branches. Bifidobacterium longum is the most abundant species in the genus Bifidobacterium. The cells of Bifidobacterium longum are long and curved, and the expanded or dumbbell-shaped rods can bifurcate; the end products produced from glucose are acetic acid and L-lactic acid. It can ferment pentoses but not gluconic acid, and is anaerobic. Bifidobacterium longum can be divided into 4 subspecies, Bifidobacterium longum subspecies, Bifidobacterium longum subspecies infantis subspecies, Bifidobacterium longum subspecies suis subspecies, and Bifidobacterium longum subspecies suilum subspecies. As a probiotic, Bifidobacterium longum not only has the characteristics of tolerance to gastric acid and bile, adhesion to intestinal epithelial cells, and inhibition of the growth of pathogenic bacteria, but also has drug resistance and does not inhibit the growth of other probiotics.

[0003] At present, the application range of probiotics is wide, distributed in many fields such as food, health products, cosmetics, and medicines. A large number of studies have shown that the health effects of probiotics are closely related to their activity. Postbiotics are a general term for the components of probiotic metabolites after probiotics are processed. Postbiotics include bacteria and metabolites. Compared with probiotics, postbiotics are safer, can withstand higher temperatures, and are easier to store and transport. The preparation process of postbiotics may lead to unstable product performance. The content of live bacteria in postbiotics is low, and the ideal probiotic performance cannot be achieved.

[0004] Chinese patent CN118059188A discloses a Bifidobacterium longum SF-B-60 postbiotic agent and its preparation method and application. The Bifidobacterium longum SF-B-60 postbiotic agent is prepared by fermenting the fermented substrate with Bifidobacterium longum SF-B-60; the fermented substrate includes the following components by weight: 1 part of licorice, 1 part of dendrobium, 1 part of wolfberry, 1 part of kudzu root, 1 part of orange peel, and 25 parts of purified water. The Bifidobacterium longum SF-B-60 postbiotic agent provided by the invention does not require the compounding of multiple strains, the preparation method is simple, regulates the function of immune cells, improves diarrhea, and effectively enhances immunity. Compared with the postbiotic agent prepared by commercially available Bifidobacterium longum and the Bifidobacterium longum SF-B-60 postbiotic agent obtained under different fermentation conditions, the Bifidobacterium longum SF-B-60 postbiotic agent provided by the invention can regulate the function of immune cells, and can also alleviate diarrhea caused by reduced immunity to a certain extent and enhance physical fitness. However, the number of viable bacteria in postbiotic preparations is relatively small.

[0005] Currently, there are few studies on postbiotic preparations of Bifidobacterium longum in the prior art. Therefore, it is crucial to screen out a strain of Bifidobacterium longum and prepare it into a corresponding postbiotic to achieve the dual effects of high content of live bacteria in the postbiotic and the ability to treat colitis. Summary of the invention

[0006] The purpose of the present invention is to provide a Bifidobacterium longum HSBL001 postbiotic, a preparation process of a composite postbiotic and an application thereof.

[0007] To achieve the above-mentioned object of the invention, the technical solution of the present invention is as follows:

[0008] In one aspect, the present invention provides a Bifidobacterium longum HSBL001, wherein the deposit number of the Bifidobacterium longum HSBL001 is CGMCC No.31244.

[0009] Specifically, the 16s rRNA sequence of Bifidobacterium longum HSBL001 is shown in SEQ ID NO:1.

[0010] SEQ ID NO: 1:

[0011] GTCTTACCATGCAGTCGAACGGGATCCATCAGGCTTTGCTTGGGGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGGGGTAATGCCGGATGTTCCAGTTGATCGCATGGTCTTCTGGGAAAGCTTTCCCGGAATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGTAACGGCCCACCGGGGCTTCAACGGGTAGCCGGCCTGAAAGGGCGACCGGCCACTTTGGGACTGAAATACGGCCCAAACTCCTACGGGAGGCAGCAGGGGGAAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACCCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGAAGCAAGCGTGAGTGAGTTTACCCGTTGAATAAGCACCGGCTAACTACGTGCCACCAGCCGCGGTAATACGTAGGGGGCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGAATGCTGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAAGCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGAAATTGACGGGGGTCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACC。

[0012] On the other hand, the present invention provides a fermentation medium for Bifidobacterium longum. The preparation method of the fermentation medium is as follows: after dissolving skim milk powder and soy powder or skim milk powder and soy protein isolate in proportion, papain is added for enzymolysis, and after inactivating the enzyme and sterilizing, a pH regulator is added to obtain the fermentation medium.

[0013] Specifically, the mass ratio of the skim milk powder and soy flour or the skim milk powder and soy protein isolate is (2-10):1;

[0014] Furthermore, the mass ratio of the skimmed milk powder to soy flour or the skimmed milk powder to soy protein isolate is (3-10):1;

[0015] Furthermore, the mass ratio of the skimmed milk powder to soy flour or the skimmed milk powder to soy protein isolate is 6:1.

[0016] Specifically, the added amount of papain is 8000-50000U / kg material.

[0017] Specifically, the conditions for enzymatic hydrolysis after adding papain are 10-60° C., 60-180 rpm, and enzymatic hydrolysis for 0.5-6 h;

[0018] Furthermore, after adding papain, the enzymatic hydrolysis conditions were 50° C., 150 rpm, and enzymatic hydrolysis for 3 h.

[0019] Specifically, the temperature of the enzyme inactivation and sterilization is 75-121°C, and the time is 10s-20min;

[0020] Furthermore, the temperature for enzyme inactivation and sterilization is 95° C. and the time is 15 minutes.

[0021] Specifically, the pH regulator is sodium citrate.

[0022] Specifically, the sodium citrate is added at a concentration of 0.75-1.5 g / L;

[0023] According to some embodiments of the present invention, the added concentration of sodium citrate is 0.75 g / L, 1 g / L, or 1.5 g / L.

[0024] Furthermore, the added concentration of the sodium citrate is 1 g / L.

[0025] Specifically, the solid content of the fermentation medium is 12%-22%;

[0026] According to some embodiments of the present invention, the solid content of the fermentation medium is 12%, 14%, 16%, or 22%.

[0027] Furthermore, the solid content of the fermentation medium is 12%.

[0028] In another aspect, the present invention provides a method for preparing the above-mentioned Bifidobacterium longum HSBL001 postbiotic, comprising inoculating the activated Bifidobacterium longum HSBL001 into the above-mentioned fermentation medium, anaerobic fermenting at 35-38°C, 120-180rpm for 18-40h, and then heat-inactivating at 75-121°C for 10s-20min to obtain the Bifidobacterium longum HSBL001 postbiotic.

[0029] Furthermore, the conditions of the anaerobic fermentation are 37° C. and 150 rpm.

[0030] Furthermore, the anaerobic fermentation time is 24-48h;

[0031] Furthermore, the anaerobic fermentation time is 24 hours.

[0032] Specifically, the culture medium used for activating Bifidobacterium longum is MRS culture medium;

[0033] According to some embodiments of the present invention, the MRS medium includes: 10.0 g / L peptone, 8.0 g / L beef extract powder, 4.0 g / L yeast extract powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 2.0 g / L diammonium hydrogen citrate, 5.0 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate and 1.0 g / L Tween 80.

[0034] According to some embodiments of the present invention, the Bifidobacterium longum is activated for 2 generations using MRS medium.

[0035] In another aspect, the present invention provides a Bifidobacterium longum HSBL001 postbiotic prepared by the above preparation method.

[0036] On the other hand, the present invention provides a method for preparing a composite postbiotic comprising the above-mentioned Bifidobacterium longum HSBL001, comprising inoculating activated Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis into the above-mentioned fermentation medium, performing anaerobic fermentation at 35-38°C and 120-180rpm, and then heat-inactivating at 75-121°C for 10s-20min to obtain a composite postbiotic of Bifidobacterium longum HSBL001.

[0037] Specifically, the ratio of the number of live bacteria of Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis is (1-6):1:(1-6).

[0038] Furthermore, the ratio of the number of live bacteria among Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis is 4:1:4.

[0039] Specifically, the Lactobacillus gasseri is Lactobacillus gasseri JZ1048.

[0040] Specifically, the Bifidobacterium adolescentis is Bifidobacterium adolescentis JZ1069.

[0041] Specifically, the conditions of the anaerobic fermentation are 37° C. and 150 rpm.

[0042] Specifically, the anaerobic fermentation time is 30-50h;

[0043] Furthermore, the anaerobic fermentation time is 40 hours.

[0044] Specifically, the culture medium used for activating Bifidobacterium longum is MRS culture medium;

[0045] According to some embodiments of the present invention, the MRS medium includes: 10.0 g / L peptone, 8.0 g / L beef extract powder, 4.0 g / L yeast extract powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 2.0 g / L diammonium hydrogen citrate, 5.0 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.04 g / L manganese sulfate and 1.0 g / L Tween 80.

[0046] According to some embodiments of the present invention, the Bifidobacterium longum is activated for 2 generations using MRS medium.

[0047] In another aspect, the present invention provides a compound postbiotic of Bifidobacterium longum HSBL001 prepared by the above preparation method.

[0048] In another aspect, the present invention provides use of the above-mentioned Bifidobacterium longum HSBL001 or Bifidobacterium longum HSBL001 postbiotic or Bifidobacterium longum HSBL001 compound postbiotic in the preparation of a medicament for preventing and / or treating colitis.

[0049] Specifically, the colitis includes infectious colitis and non-infectious colitis;

[0050] Furthermore, the infectious colitis includes bacterial colitis and viral colitis;

[0051] Furthermore, the non-infectious colitis includes ulcerative colitis and Crohn's disease.

[0052] Furthermore, the colitis is ulcerative colitis.

[0053] Specifically, the drug may also include a pharmaceutically acceptable carrier, and the pharmaceutically acceptable carrier is selected from one or more of excipients, buffers, emulsifiers, stabilizers, diluents, adhesives, preservatives, lubricants, pH regulators, cryoprotectants, and flavoring agents.

[0054] Specifically, the dosage form of the drug is tablets, powders, oral liquids, capsules, granules, ointments, suspensions, pills, and lozenges.

[0055] The beneficial effects of the present invention are:

[0056] 1. The culture medium optimized by the present invention is obtained by enzymatic hydrolysis of skim milk and soybean powder with protease, which solves the problem of difficulty in fermentation of Bifidobacterium longum in skim milk. It not only takes into account the taste, but also greatly increases the number of live fermented bacteria of Bifidobacterium longum in the skim milk base, reaching 5.2 billion CFU / mL.

[0057] 2. The combined fermentation of Bifidobacterium longum with Lactobacillus gasseri and Bifidobacterium adolescentis can produce synergistic fermentation and effectively increase the total number of live bacteria.

[0058] 3. The postbiotics of Bifidobacterium longum HSBL001 and the postbiotics of the combined fermentation of Bifidobacterium longum HSBL001 with Lactobacillus gasseri JZ1048 and Bifidobacterium adolescentis JZ1069 have the effect of relieving colitis.

[0059] Deposit certificate

[0060] Deposit number: CGMCC No.31244;

[0061] Classification and nomenclature: Bifidobacterium longum;

[0062] Preservation time: July 9, 2024;

[0063] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0064] Abbreviation of depository unit: CGMCC;

[0065] Collection address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 The effects of HSBL001 postbiotics and HSBL001 compound postbiotics on animal food intake (A), body weight (B), disease activity index (C) and spleen index (D). In the figure, "*" indicates a significant difference compared with the model group, P < 0.05.

[0067] Figure 2The effect of HSBL001 postbiotics and HSBL001 compound postbiotics on the colon length of mice. In the figure, "*" indicates a significant difference compared with the model group, P < 0.05. DETAILED DESCRIPTION

[0068] In order to make the technical means, creative features, purpose and efficacy of the present invention easy to understand, the present invention is further explained in conjunction with specific embodiments below, but the following embodiments are only preferred embodiments of the present invention, not all. Based on the embodiments in the implementation mode, other embodiments obtained by those skilled in the art without creative work all belong to the protection scope of the present invention. In the following embodiments, unless otherwise specified, the operating methods used are conventional operating methods, the equipment used are conventional equipment, and the equipment materials used in each embodiment are the same.

[0069] Source of experimental strains:

[0070] Lactobacillus gasseri JZ1048, Lactobacillus reuteri JZ1039, Lactobacillus bulgaricus JZ1045, Streptococcus thermophilus JZ1041, Bifidobacterium bifidum JZ1025, Bifidobacterium longum infantis subsp. JZ1030, Bifidobacterium adolescentis JZ1069, Bifidobacterium infantis JZ1076, Bifidobacterium adolescentis JZ1068, Bifidobacterium infantis JZ1070, and Lactobacillus bulgaricus JZ1045 were provided by Jiangzhong Pharmaceutical Co., Ltd. All strains were isolated and identified from samples.

[0071] Example 1 Optimizing the stirring process to increase the number of viable bacteria of Bifidobacterium longum HSBL001 in skim milk base

[0072] The glycerol tube of Bifidobacterium longum HSBL001 was activated for 2 generations with MRS broth medium (10.0 g of peptone, 8.0 g of beef extract powder, 4.0 g of yeast extract powder, 20.0 g of glucose, 2.0 g of dipotassium hydrogen phosphate, 2.0 g of diammonium hydrogen citrate, 5.0 g of sodium acetate, 0.2 g of magnesium sulfate, 0.04 g of manganese sulfate and 1.0 g of Tween 80 were dissolved and fixed to 1 L of water), and the bacterial liquid was centrifuged to obtain bacterial mud, which was resuspended with an equal amount of physiological saline for use. Skim milk medium was weighed according to 14% (w / w) skim milk powder, dissolved in water, and fixed to volume with water, and then transferred to a fermenter for cooling after sterilization.

[0073] 3% (v / v) of the resuspended bacterial solution was inoculated into a 14% skim milk medium, and anaerobically fermented at 150rpm and 37°C for 30 hours, with static fermentation as a control. The results are shown in Table 1 below. It can be seen from the results that the number of viable bacteria in the stirred fermentation was significantly increased, and the fermentation acidity was significantly higher than that in the static fermentation.

[0074] Table 1 Effect of stirring on viable bacterial count and acidity

[0075] <![CDATA[Viable count × 10 8 CFU / g]]> Acidity °T Static fermentation 1.80 103.00 150 rpm stirring 3.43 130.50

[0076] Example 2 Compounding of fermentation substrates to increase the number of viable bacteria of Bifidobacterium longum HSBL001 in skim milk base

[0077] Using 14% skim milk as the basic culture medium, different proportions of soy flour, soy protein isolate, or whey protein were mixed with skim milk as substrates to ferment Bifidobacterium longum. The inoculation amount of Bifidobacterium longum was 3%, and the fermentation was carried out at 37℃ for 30h. From the results, it can be seen that the number of live bacteria increased to a certain extent after compounding, among which the number of live bacteria in the formula of 8% soy protein isolate + 6% skim milk was the highest, which was 13.17×10 8 CFU / g, but the taste was poor after adding soy protein isolate. The second was whey protein 14%, with a viable count of 13.20×10 8 CFU / g, but the taste is poor.

[0078] Table 2 Effects of soy flour, soy protein isolate and whey protein powder combined with skim milk on the number of viable Bifidobacterium longum and acidity

[0079]

[0080]

[0081] Example 3 Adding buffer salt to increase the number of viable bacteria of Bifidobacterium longum HSBL001 in skim milk base

[0082] After Bifidobacterium longum HSBL001 was activated for 2 generations with MRS medium, it was inoculated into soybean powder-skim milk medium (soy powder 2% + 12% skim milk) containing different concentrations of sodium citrate at a 3% inoculation amount, and the viable bacteria were counted after anaerobic culture at 37°C and 150rpm for 24 hours. The results show that the initial pH value of the culture medium can be increased after adding sodium citrate, and the viable bacteria count is the highest when the addition amount is 1.0g / L. The viable bacteria count tends to decrease as the concentration of sodium citrate is further increased.

[0083] Table 3 Effect of sodium citrate on the number of viable cells of Bifidobacterium longum HSBL001 fermentation

[0084] Addition concentration g / L Initial pH <![CDATA[Viable count × 10 8 CFU / g]]> 0 6.61 30.27 0.75 6.75 32.80 1.0 6.76 33.53 1.5 6.81 32.80 2.0 6.86 31.33 2.5 6.88 30.17 3.0 6.86 27.87

[0085] Example 4 Optimizing the enzymatic hydrolysis process to increase the number of viable bacteria of Bifidobacterium longum HSBL001 in skim milk base

[0086] In order to further increase the number of viable bacteria in fermentation, different proteases were used to enzymolyze the culture medium of 2% soybean powder + 12% skim milk. Papain, flavor protease, etc. were enzymolyzed according to the optimal addition amount and enzymolysis process selected above. The addition amount is shown in the table. The enzymolysis conditions are uniformly 50℃ for 3h. After the enzymolysis is completed, the enzyme is quickly sterilized in an autoclave at 95℃ for 10min. After anaerobic fermentation at 37℃ and 150rpm for 24h after inoculation, it can be seen from the results that papain has the highest number of viable bacteria, which is 52.0×10 8 CFU / g, and no odor.

[0087] Table 4 Effects of different enzymatic hydrolysis processes on the fermentation of Bifidobacterium longum in skim milk base

[0088]

[0089]

[0090] Example 5 Optimizing the solid content of the culture medium to increase the number of viable bacteria of Bifidobacterium longum HSBL001 fermented in skim milk base

[0091] The ratio of the fixed culture medium formula is soybean powder: skim milk = 1:6, and the solid content is set to 12%-22%. After inoculation, anaerobic fermentation is carried out at 37℃ and 150rpm for 24 hours, and the number of viable bacteria in the fermentation is detected. It can be seen that increasing the solid content can increase the number of viable bacteria in the fermentation. When the solid content is 22%, the number of viable bacteria is the highest, which is 66×10 8 CFU / g. Spray drying was performed after fermentation to calculate the theoretical bacterial count (theoretical bacterial count = number of viable bacteria / solid content). From the results in the table below, it can be seen that increasing the fermentation solid content will reduce the theoretical bacterial count in the postbiotics, and it is relatively high at 16%.

[0092] Table 5 Effect of different solid content on viable count

[0093] Solid content <![CDATA[Number of viable bacteria × 10 8 CFU / g]]> <![CDATA[Theoretical number of bacteria cells × 10 8 CFU / g]]> 12％ 40 333.3 14％ 41 292.9 16％ 51 318.8 18％ 51 283.3 22％ 66 300.0

[0094] Example 6: Compounding of two bacteria to increase total viable count

[0095] Bifidobacterium longum HSBL001 was used as the main fermentation strain, and other strains were compounded to increase the number of viable fermentation bacteria and the content of metabolites. The initial viable number of Bifidobacterium longum HSBL001 was controlled to be 0.4×10 8 CFU / g, the initial viable count of other strains was 0.1×10 8CFU / g, after inoculation, anaerobic culture was carried out at 37℃150rpm for 24h before viable count. From the results of total viable count (Table 6), it can be seen that the total viable count of Lactobacillus gasseri JZ1048 is higher, followed by Lactobacillus reuteri JZ1039. After compounding with the four strains of Bifidobacterium tested, the total viable count was significantly improved, among which the total viable count of compounding with Bifidobacterium adolescentis JZ1069 was the highest.

[0096] Table 6 Study on the combination of two of Bifidobacterium longum HSBL001

[0097]

[0098]

[0099] Example 7 Study on the multi-bacterial compound of Bifidobacterium longum HSBL001

[0100] Through the screening of two bacteria compounding, it was found that the compound fermentation of Bifidobacterium longum HSBL001 with Lactobacillus reuteri JZ1039 and Lactobacillus gasseri JZ1048 had a higher total viable count and retained more Bifidobacterium longum HSBL001. On this basis, Bifidobacterium bifidum was further compounded to increase the total viable count and the number of bifidobacteria. The initial viable count of Bifidobacterium longum HSBL001 was controlled to be 0.4×10 8 CFU / g, the ratio of Bifidobacterium longum HSBL001 to other strains was 4:1:4, and the viable bacteria were counted after anaerobic culture at 37℃150rpm for 24h after inoculation. It can be seen from the results (Table 7) that the total number of viable bacteria in the combination group of Bifidobacterium longum HSBL001 and Lactobacillus gasseri JZ1048 was higher than that in the combination group of Lactobacillus reuteri JZ1039 and Lactobacillus bulgaricus JZ1045. Among them, after the combination fermentation of Bifidobacterium longum HSBL001+Lactobacillus gasseri JZ1048+Bifidobacterium adolescentis JZ1069, the total number of viable bacteria and the total number of bifidobacteria were both higher, indicating that bifidobacteria and Lactobacillus gasseri JZ1048 can better cooperate in fermentation.

[0101] Table 7 Study on multi-bacteria combination of Bifidobacterium longum HSBL001

[0102]

[0103]

[0104]

[0105] Example 8 Effects of Bifidobacterium longum HSBL001 postbiotics and Bifidobacterium longum HSBL001 combined postbiotics on dextran sodium sulfate-induced colitis in mice

[0106] 1 Experimental methods

[0107] (1) Animal husbandry and grouping: After 7 days of adaptive feeding, C57bl / 6 male mice were randomly divided into groups according to their body weight (normal group, model group, HSBL001 postbiotic group, and HSBL001 combined postbiotic group), with 8 mice in each group.

[0108] The preparation method of HSBL001 postbiotics is as follows: 13.7% skim milk, 2.3% soy flour, and 84% water are mixed evenly, 20000U / kg of papain is added, and enzymolysis is carried out at 50℃ for 3h. After the enzymolysis is completed, 1.0g / L sodium citrate is added, and the enzyme is quickly sterilized in an autoclave at 95℃ for 10min. Bifidobacterium longum HSBL001 is inoculated to make the initial viable count of 0.4×10 8 CFU / g, anaerobic culture at 37℃150rpm for 24h, heat inactivation at 95℃ for 15min. HSBL001 postbiotics were obtained after spray drying.

[0109] The preparation method of HSBL001 compound postbiotics is as follows: skim milk 13.7%, soy flour 2.3%, water 84% are mixed evenly, papain is added in an amount of 50000U / kg, enzymolysis is carried out at 50℃ for 3h, 1.0g / L sodium citrate is added after the enzymolysis is completed, and the enzyme is quickly sterilized in a sterilizer at 95℃ for 10min. Bifidobacterium longum HSBL001, Lactobacillus gasseri JZ1048 and Bifidobacterium adolescentis JZ1069 are inoculated, and the initial viable counts are controlled to be 0.4×10 8 , 0.1×10 8 and 0.4×10 8 CFU / g, anaerobic culture at 37℃150rpm for 24h, heat inactivation at 95℃ for 15min. HSBL001 compound postbiotics were obtained after spray drying.

[0110] (2) Modeling and drug administration: After the start of the experiment, the samples were gavaged every day. The HSBL001 postbiotic group and the HSBL001 compound postbiotic group were gavaged with 1.0 g / kg (bw) of the sample every day (human equivalent dose 4.8 g / day), and the normal group and the model group were gavaged with an equal volume (0.2 mL) of PBS every day. On the 8th day of intervention, DSS solution (2.5%) was used as the drinking water source for modeling, and the samples were gavaged for intervention at the same time, and the modeling continued for 7 days.

[0111] (3) Impact on animal disease activity index

[0112] On the 14th day, the experimental mice were housed in single cages, and the disease activity index (DAI) was scored after the mice defecated. The DAI value was [DAI = (weight loss score + rectal bleeding score + fecal characteristics score) / 3].

[0113] DAI evaluation criteria refer to the literature, as follows:

[0114] The proportion of body weight loss (0: no weight loss; 1: 1%-5%; 2: 5%-10%; 3: 10%-20%; 4: >20%).

[0115] Stool blood (0 = no blood; 1 = slightly bloody; 2 = some bloody; 3 = quite a bit of bloody; 4 = a lot of bloody).

[0116] Stool consistency (0 = normal stool; 1 = slightly loose stool; 2 = loose stool; 3 = watery stool; 4 = very loose stool).

[0117] From the results, it can be seen that ( Figure 1 ), the disease activity index of mice gavaged with HSBL001 postbiotics and HSBL001 compound postbiotics was significantly lower than that of the model group, and the diarrhea symptoms of the mice were alleviated, with basically no blood in the stool.

[0118] (4) Effect on colon length: The colon length of mice was measured after fasting but not water restriction for 12 hours starting from the 14th day. The results showed that after DSS modeling, the colon of mice was significantly shorter than that of the blank group, indicating that the modeling was successful. After intragastric administration of postbiotic samples, the colon length was restored to a certain extent, and there was a significant difference compared with the model group ( Figure 2 ).

[0119] (5) Effect on spleen index: After the mice were dissected, their spleens were weighed and spleen index was calculated, spleen index = spleen mass m1 / body weight m. The results show that after DSS modeling, the spleen index of the model group was significantly higher than that of the blank group, the spleen was enlarged, and the digestion and metabolism of the mice were abnormal. After intervention with postbiotic samples, the phenomenon of spleen enlargement was significantly restored ( Figure 1 ).

[0120] 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 principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A Bifidobacterium longum HSBL001, characterized in that The deposit number of the Bifidobacterium longum HSBL001 is CGMCC No.31244.

2. The fermentation medium of Bifidobacterium longum HSBL001 according to claim 1, characterized in that The preparation method of the fermentation medium is as follows: after dissolving skim milk powder and soybean powder or skim milk powder and soybean protein isolate, papain is added for enzymolysis, and after inactivating the enzyme and sterilizing, a pH regulator is added to obtain the fermentation medium.

3. The fermentation medium according to claim 2, characterized in that The mass ratio of the skimmed milk powder to soybean powder or the skimmed milk powder to soybean protein isolate is (2-10):

1.

4. The fermentation medium according to claim 2, characterized in that The added amount of the papain is 8000-50000U / kg.

5. The fermentation medium according to claim 2, characterized in that The pH regulator is sodium citrate, and the added concentration of the sodium citrate is 0.75-1.5 g / L.

6. The fermentation medium according to claim 2, characterized in that The solid content of the fermentation medium is 12%-22%.

7. A method for preparing a postbiotic comprising the Bifidobacterium longum HSBL001 according to claim 1, characterized in that: The activated Bifidobacterium longum HSBL001 is inoculated into the fermentation medium according to any one of claims 2 to 6, and after anaerobic fermentation at 35-38° C. and 120-180 rpm for 18-40 hours, heat inactivation is performed at 75-121° C. for 10 seconds to 20 minutes to obtain Bifidobacterium longum HSBL001 postbiotics.

8. The Bifidobacterium longum HSBL001 postbiotic prepared by the preparation method according to claim 7.

9. A method for preparing a composite postbiotic comprising the Bifidobacterium longum HSBL001 according to claim 1, characterized in that: The activated Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis are inoculated into the fermentation medium according to any one of claims 2 to 6, and anaerobically fermented at 35-38° C. and 120-180 rpm, and then heat-inactivated at 75-121° C. for 10 s to 20 min to obtain the product.

10. The preparation method according to claim 9, characterized in that: The ratio of the number of live bacteria among Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis is (1-6):1:(1-6).

11. The preparation method according to claim 10, characterized in that: The ratio of the number of live bacteria among Bifidobacterium longum HSBL001, Lactobacillus gasseri and Bifidobacterium adolescentis is 4:1:

4.

12. The composite postbiotic of Bifidobacterium longum HSBL001 prepared by the preparation method according to any one of claims 9 to 11.

13. Use of the Bifidobacterium longum HSBL001 according to claim 1 or the Bifidobacterium longum HSBL001 postbiotic according to claim 8 or the Bifidobacterium longum HSBL001 composite postbiotic according to claim 12 in the preparation of a medicament for preventing and / or treating colitis.

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