Preparation process of metabiotics and functional application of metabiotics for relieving constipation

Abstract

The invention discloses a preparation process of metabiotics and functional application of the metabiotics for relieving constipation, and relates to the technical field of microorganisms. The invention provides a preparation method of a bifidobacterium animalis subsp. Lactis LIHUO 01-containing metagen, which comprises the following steps: separately inoculating bifidobacterium animalis subsp. Lactis LIHUO 01 or compounding with probiotics to inoculate in a fermentation culture medium for culturing to obtain a fermentation bacteria solution, and carrying out thermal inactivation to obtain the bifidobacterium animalis subsp. Lactis LIHUO 01-containing metagen. The fermentation medium comprises skimmed milk, soybean meal and water, the mass ratio of the skimmed milk to the soybean meal to the water is (5-30): (1-25): (78-90), and the prepared bifidobacterium animalis subsp. Lactis LISUO 01 metabolite can significantly improve the content of tryptophan metabolites, can effectively relieve constipation and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of microorganisms, in particular to a postbiotic preparation process and its application in relieving constipation. BACKGROUND

[0002] Probiotics are a class of active microorganisms that are beneficial to the host, are planted in the human intestinal tract and reproductive system, can produce definite health effects to improve the microecological balance of the host and play a beneficial role. Lactic acid bacteria, as the main strain of probiotics, participate in the action of intestinal microorganisms, and their metabolites such as acetic acid have an important role in the health of the host. Bifidobacterium is a non-spore, non-motile, trans-anaerobic, gram-positive bacterium. The cell morphology of bifidobacterium is diverse, including short rod, near spherical, fiber rod, bifurcated rod, stick or spoon. The most common form is rod. In morphology, bifidobacterium is mainly divided into two types, type I and type II. Type I refers to the cell morphology of bifurcation, and type II refers to the cell morphology of rod.

[0003] Postbiotics refers to inactivated microorganisms and / or bacterial components with a clear genetic background that are beneficial to the health of the host, including or not including their metabolites. The main components include inactivated bacterial cells, bacterial components and bacterial metabolites, wherein the bacterial components include lipoteichoic acid, peptidoglycan, cell surface protein, etc., and the bacterial metabolites include extracellular polysaccharide, short-chain fatty acid, protein / polypeptide, etc.

[0004] Constipation is a common clinical gastrointestinal disease characterized by difficulty in defecation, low frequency or incomplete defecation. The main causes of constipation are as follows: (1) lifestyle or dietary habits: long sitting, lack of exercise, insufficient water intake, and lack of dietary fiber in diet; (2) intestinal microbial disorder: changes in the composition and number of intestinal microorganisms affect the colonization and metabolism of microbial populations. There are many causes of constipation, and the etiology is complex. Long-term constipation symptoms can significantly reduce the quality of life of patients. The formation mechanism of constipation includes the following: (1) the intestinal peristalsis ability is weakened; (2) the intestinal microecological homeostasis is disturbed, and the disturbance of intestinal flora can cause the disorder of bile acid metabolism, and the disorder of bile acid metabolism can cause the relaxation and contraction disorder of the proximal colon, thereby affecting the colon peristalsis and causing constipation or diarrhea in the host; (3) abnormal water transport in the intestine; (4) immune balance of the intestine.

[0005] Tryptophan metabolites, such as indole-3-lactic acid and indole-3-acetic acid, have a positive effect on promoting intestinal health, but the strains producing them are mainly intestinal bacteria, and there are few reports on food-grade probiotics producing tryptophan metabolites. Therefore, it is urgent to provide a preparation method of probiotic postbiotics with high content of tryptophan metabolites and capable of relieving constipation. SUMMARY

[0006] The application aims to provide a postbiotic preparation process and its application in relieving constipation.

[0007] To achieve the above-mentioned application purposes, the technical solutions of the application are as follows. In one aspect, the application provides a preparation method of a postbiotic containing Bifidobacterium animalis lactis LIHUO 01, the preservation number of the Bifidobacterium animalis lactis LIHUO 01 being GDMCC No:65512, the preparation method being inoculating the Bifidobacterium animalis lactis LIHUO 01 alone or inoculating the Bifidobacterium animalis lactis LIHUO 01 and probiotics in a fermentation medium to obtain fermentation broth, and then performing heat inactivation to obtain the postbiotic of the Bifidobacterium animalis lactis LIHUO 01. The fermentation medium comprises skimmed milk, soybean powder and water, and the mass ratio of the skimmed milk, the soybean powder and the water is (5-30):(1-25):(78-90). The probiotics are selected from one or more of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri and Lactobacillus gasseri.

[0008] Further, the probiotics are selected from Lactobacillus plantarum or Lactobacillus gasseri.

[0009] Specifically, the ratio of the Bifidobacterium animalis lactis LIHUO 01 to the Lactobacillus plantarum or the Lactobacillus gasseri is 1:1.

[0010] Further, the Lactobacillus plantarum is Lactobacillus plantarum P9. Further, the Lactobacillus gasseri is Lactobacillus gasseri JZ60.

[0011] Further, the mass ratio of the skimmed milk, the soybean powder and the water in the fermentation medium is 12:2:86.

[0012] Specifically, the fermentation medium further comprises a protease.

[0013] Further, the addition amount of the protease accounts for 10-1000 U / g of the dry matter content. Further, the addition amount of the protease accounts for 350 U / g of the dry matter content.

[0014] Specifically, the protease is selected from one or more of papain, flavour protease, acid protease, neutral protease, bromelain and alkaline protease.

[0015] Further, the protease is selected from one or more of papain, flavour protease, neutral protease and bromelain.

[0016] Specifically, the enzymolysis conditions after adding the protease are 40-60℃ for 2-4h; Further, the enzymolysis conditions after adding the protease are 50℃ for 3h.

[0017] Specifically, the postbiotic containing Bifidobacterium animalis lactis LIHUO 01 in the present application comprises dead cells and / or bacterial lysates and / or strain metabolites.

[0018] Further, the strain metabolites comprise indole-3-lactic acid, indole-3-acetic acid, indole-3-propionic acid, indole-3-formaldehyde, lactic acid, succinic acid and acetic acid.

[0019] Specifically, the inoculation amount of Bifidobacterium animalis lactis LIHUO 01 is 1-10% (v / v); Further, the inoculation amount of Bifidobacterium animalis lactis LIHUO 01 is 1-5% (v / v); Still further, the inoculation amount of Bifidobacterium animalis lactis LIHUO 01 is 3%.

[0020] Specifically, the culture conditions of Bifidobacterium animalis lactis LIHUO 01 are 35-38℃ anaerobic culture at 100-200rpm for 24-48h; Further, the culture conditions of Bifidobacterium animalis lactis LIHUO 01 are 37℃ anaerobic culture at 150rpm for 48h.

[0021] Specifically, the heat inactivation conditions are 70-120℃ heat inactivation for 0.5-30min; Further, the heat inactivation conditions are 105℃ heat inactivation for 5min.

[0022] In another aspect, the present application provides a postbiotic containing Bifidobacterium animalis lactis LIHUO 01 prepared by the above preparation method.

[0023] In another aspect, the present application provides a fermentation medium, wherein the fermentation medium comprises skimmed milk, soybean powder and water, and the mass ratio of skimmed milk: soybean powder: water is (5-30):(1-25):(78-90).

[0024] Further, the mass ratio of skimmed milk: soybean powder: water in the fermentation medium is 12:2:86.

[0025] Specifically, the fermentation medium further comprises a protease.

[0026] Further, the addition amount of the protease accounts for 10-1000U / g of the dry matter content. Still further, the added amount of the protease is 350 U / g of dry matter content.

[0027] Specifically, the protease is selected from one or more of papain, flavourzyme, acid protease, neutral protease, bromelain, and alkaline protease.

[0028] Further, the protease is selected from one or more of papain, flavourzyme, neutral protease, and bromelain.

[0029] In another aspect, the present application provides use of the above-mentioned animal bifidobacterium lactis LIHUO 01 probiotic in the preparation of a product for relieving constipation.

[0030] Specifically, the product includes but is not limited to pharmaceuticals, food, and animal nutrition products.

[0031] Specifically, the food includes general food, special medical purpose food, and health food.

[0032] Further, the product is a pharmaceutical.

[0033] Specifically, when the product is a pharmaceutical, the pharmaceutical can further include pharmaceutically acceptable raw materials and excipients.

[0034] Further, the pharmaceutical excipient is selected from any one or more of adjuvants, stabilizers or protective agents, bacteriostatic agents, excipients, solubilizers, flavoring agents, diluents, buffers.

[0035] Specifically, the dosage form of the pharmaceutical is any one or more of tablets, capsules, granules, eye preparations, nasal preparations, suppositories, ointments, creams, sprays, gels, powders, coatings, lyophilized agents.

[0036] Further, the food includes but is not limited to pressed candies, yogurts, cans, biscuits, chocolates, pastries, creams, cheeses, cream cheeses, milk powders, ice creams, ice pops, fruit jams, fruit purees, candied fruits, preserved fruits, dried fruits, breads, egg rolls, protein beverages, lactic acid bacteria beverages, plant protein beverages, carbonated beverages, coffee, puffed foods.

[0037] Further, when the product is general food or health food or animal nutrition products, the general food or health food further includes additives, and the additives are selected from any one or more of acidity regulators, anti-caking agents, antifoaming agents, antioxidants, bleaching agents, bulking agents, coloring agents, color protection agents, emulsifiers, enzyme preparations, flavor enhancers, film forming agents, moisture retention agents, nutritional fortifiers, preservatives, stabilizers and coagulants, sweeteners, thickening agents, food flavorings, and processing aids for the food industry.

[0038] Further, the health food includes emulsion products, solution products, powder products and solid products.

[0039] Specifically, the product has the following effects: (1) increasing the water content of feces; (2) improving the intestinal peristalsis and accelerating the small intestine propulsion rate; (3) shortening the defecation time.

[0040] In another aspect, the present application provides a product for relieving constipation, wherein the product comprises the above-mentioned animal bifidobacterium lactis LIHUO 01 probiotic.

[0041] Specifically, the product includes but is not limited to pharmaceuticals, food and animal nutrition products.

[0042] Specifically, the food includes general food, special medical purpose food and health food.

[0043] Specifically, the product helps to moisten the intestines and defecate.

[0044] Further, the product is a pharmaceutical.

[0045] Specifically, when the product is a pharmaceutical, the pharmaceutical can further comprise pharmaceutically acceptable raw materials and excipients.

[0046] Further, the pharmaceutically acceptable carrier is selected from any one or more of adjuvants, stabilizers or protective agents, bacteriostatic agents, excipients, solubilizers, flavoring agents, diluents, buffers.

[0047] Specifically, the dosage form of the pharmaceutical is any one or more of tablets, capsules, granules, eye preparations, nasal preparations, suppositories, ointments, creams, sprays, gels, powders, paints, lyophilized agents.

[0048] Further, the food includes but is not limited to pressed tablet candies, yogurts, cans, biscuits, chocolates, pastries, butters, cheeses, cream cheeses, milk powders, ice creams, ice creams, fruit jams, fruit purees, candied fruits, preserved fruits, breads, egg rolls, egg white beverages, lactic acid bacteria beverages, plant protein beverages, carbonated beverages, coffee, puffed foods.

[0049] Further, the product is a common food or a health food or an animal nutrition product, and the common food or the health food further comprises an additive selected from any one or more of an acidity regulator, an anti-caking agent, an antifoaming agent, an antioxidant, a bleaching agent, a bulking agent, a coloring agent, a color protection agent, an emulsifying agent, an enzyme preparation, a flavor enhancer, a film forming agent, a moisture retaining agent, a nutrient fortifier, a preservative, a stabilizer and a coagulating agent, a sweetener, a thickening agent, a food flavor, and a processing aid for the food industry.

[0050] Further, the health food comprises an emulsion product, a solution product, a powder product, and a solid product.

[0051] In another aspect, the present application provides a use of the above-mentioned animal Bifidobacterium lactis LIHUO 01 postbiotic in the preparation of a microbial preparation for producing indole-3-lactic acid, indole-3-acetic acid, or short-chain fatty acid.

[0052] The present application has the following advantages: (1) The preparation process of the animal Bifidobacterium lactis LIHUO 01 postbiotic can significantly increase the content of tryptophan metabolites.

[0053] (2) Compared with the loperamide-induced constipated mice, the mice administered with the animal Bifidobacterium lactis LIHUO 01 postbiotic have a shorter defecation time. The postbiotic has the effect of shortening the defecation time and has the effect of relieving, improving, or treating functional constipation. BRIEF DESCRIPTION OF DRAWINGS

[0054] Figure 1 For the influence on the water content of animal feces, "*" indicates a significant difference from the model group, p <0.05.

[0055] Figure 2 For the influence on the length of the small intestine and the intestinal propulsion rate of the animals, "*" indicates a significant difference from the model group, p <0.05.

[0056] Figure 3 For the influence on the serum motilin (MLT) level of the animals, "*" indicates a significant difference from the model group, p <0.05. DETAILED DESCRIPTION

[0057] In order to make the technical means, creative features, purposes and effects of the present application easy to understand, the following specific embodiments are further described to clarify the present application. However, the following embodiments are only preferred embodiments of the present application, not all. Based on the embodiments in the embodiments, other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application. In the following examples, if not otherwise specified, the operation method used is a conventional operation method, the equipment used is a conventional equipment, and the equipment materials used in each example are the same.

[0058] The animal bifidobacterium lactis LIHUO 01 described in the present application has been disclosed in patent CN119331785A.

[0059] The plant lactobacillus P9 has a preservation number of CGMCC No.16662, which has been disclosed in patent CN113558244A.

[0060] Example 1: Effect of different culture substrates on the production of tryptophan metabolites by animal bifidobacterium lactis LIHUO 01 (1) Strain activation: After 2 generations of activation of the glycerol tube of animal bifidobacterium lactis LIHUO 01, the bacterial slurry was obtained by centrifugation of the bacterial liquid, and the bacterial slurry was resuspended with an equal amount of normal saline for standby.

[0061] (2) Preparation of culture medium: BBL liquid medium was prepared according to the instruction manual and sterilized. The skim milk medium was prepared by weighing 14% (w / w) of skim milk powder, dissolving it in water, and then adding water to constant volume. After sterilization, it was transferred to the fermentation tank for cooling standby. The skim milk soybean powder medium, skim milk soybean protein isolate medium and skim milk whey protein medium were prepared according to the ratio of skim milk: soybean powder (soybean protein isolate or whey protein): water = 12:2:86 (w / w), dissolved in water, and then added with water to constant volume. After sterilization, it was transferred to the fermentation tank for cooling standby.

[0062] (3) Inoculation and fermentation: The resuspended bacterial liquid was inoculated into different culture media at a 3% (v / v) inoculation amount, and cultured at 37°C under anaerobic conditions at 150 rpm for 48 h.

[0063] (4) Tryptophan metabolites detection: Indole-3-lactic acid, indole-3-acetic acid and indole-3-propionic acid standard solutions were prepared by dissolving the standard samples in methanol at concentrations of 87.0 mg / L, 76.7 mg / L and 79.4 mg / L, respectively, and then gradient dilution and filtration. The liquid chromatography conditions were as follows: Agilent ZORBAX SB-C18 column, flow rate 1.0 m L / min, sample volume 10 L, detection wavelength 250 nm and 280 nm, mobile phase acetonitrile (A): 0.1% formic acid water (B) gradient elution, 0-5 min: 5% A, 95% B; 5-20 min: 5%-20% A, 95%-80% B; 20-40 min: 20% A, 80% B; 40-60 min: 20%-40% A, 80%-60% B.

[0064] The detection results are shown in Table 1. It can be seen from the results that different culture medium substrates have a significant effect on the production of tryptophan metabolites by Bifidobacterium animalis lactis LIHUO 01. The production of indole-3-lactic acid in the skim milk-soybean flour medium is significantly higher than that in other media.

[0065] Table 1 Effect of different culture substrates on the production of tryptophan metabolites by Bifidobacterium animalis lactis LIHUO 01

[0066] Note: letters represent significance p <0.05.

[0067] Example 2 Effect of different protease enzymolysis on the production of tryptophan metabolites by Bifidobacterium animalis lactis LIHUO 01 The control group medium was prepared according to the ratio of skim milk: soybean flour: water = 12:2:86 (w / w). Different enzyme groups were added to the control group medium at a dosage of 350 U / g dry matter, and enzyme hydrolysis was carried out at 50°C for 3h. After enzyme hydrolysis, the medium was sterilized by enzyme inactivation at 105°C. After activation, the Bifidobacterium animalis lactis LIHUO 01 was resuspended with physiological saline after centrifugation, and inoculated into different media at an inoculation amount of 3% (v / v), and cultured at 37°C under anaerobic conditions at 150 rpm for 48h.

[0068] As can be seen from the results (Table 2), the substrate after different protease enzymolysis has a great effect on the production of tryptophan metabolites by Bifidobacterium animalis lactis LIHUO 01. The six tested proteases can significantly increase the content of indole-3-lactic acid. After neutral protease enzymolysis, the indole-3-lactic acid increased to 26.743 mg / L. Alkaline protease increased indole-3-propionic acid most significantly, which was 7.6 times higher than the control group.

[0069] Table 2 Effect of enzymolysis on tryptophan metabolites produced by Bifidobacterium animalis subsp. lactis LIHUO 01

[0070] Note: "*" indicates a significant difference compared with the control group, p <0.05.

[0071] Example 3 Preparation of animal Bifidobacterium animalis subsp. lactis LIHUO 01 postbiotic sample Preparation of postbiotic sample 1: After 2 generations of activation of Bifidobacterium animalis subsp. lactis LIHUO 01 according to the above method, the bacterial slurry was collected by centrifugation at 6000 rpm for 5 min, washed twice with normal saline, resuspended with an equal amount of normal saline, and stored for use. The skim milk-soybean powder medium was prepared according to the ratio of skim milk: soybean powder: water = 12:2:86 (w / w), dissolved thoroughly with warm water, and then 350 U / g of dry matter of neutral protease was added for enzymolysis at 50°C for 3 h. Immediately after the end of enzymolysis, the enzyme was inactivated and sterilized at 105°C. After cooling, the resuspended bacterial solution was inoculated at an inoculation amount of 3% (v / v) and anaerobically fermented at 37°C for 40 h. The fermented sample was detected for relevant physical and chemical indicators, and the results are shown in Table 3. The viable bacterial count in the sample was 4.63 billion CFU / g, and the fermentation acidity was 173.5ºT.

[0072] The fermentation broth was heat-inactivated at 105°C for 5 min, homogenized with an emulsifier, and then spray-dried at an inlet temperature of 170°C and an outlet temperature of 80°C, with a feeding speed of 12 rpm and an atomizer speed of 18000 rpm. Thus, the animal Bifidobacterium animalis subsp. lactis LIHUO 01 postbiotic sample 1 was obtained.

[0073] Preparation of postbiotic sample 2: The difference from postbiotic sample 1 is only in the preparation method of the fermentation medium. The skim milk-soybean powder medium was prepared according to the ratio of skim milk: soybean powder: water = 5:1:78 (w / w), dissolved thoroughly with warm water, and then 90 U / g of dry matter of papain was added for enzymolysis at 50°C for 3 h. Immediately after the end of enzymolysis, the enzyme was inactivated and sterilized at 105°C.

[0074] Preparation of postbiotic sample 3: The difference from postbiotic sample 1 is only in the preparation method of the fermentation medium. The skim milk-soybean powder medium was prepared according to the ratio of skim milk: soybean powder: water = 30:25:90 (w / w), dissolved thoroughly with warm water, and then 700 U / g of dry matter of alkaline protease was added for enzymolysis at 50°C for 3 h. Immediately after the end of enzymolysis, the enzyme was inactivated and sterilized at 105°C.

[0075] Preparation of postbiotic sample 4: The difference between the postbiotic sample 1 is only in the preparation method of the fermentation medium, the skim milk medium is prepared according to skim milk: water = 20:80 (w / w), after fully dissolving with warm water, adding alkaline protease 700 U / g dry matter, enzyme hydrolysis at 50℃ for 3h, immediately after enzyme hydrolysis, enzyme inactivation and sterilization at 105℃.

[0076] Preparation of postbiotic sample 5: The difference between the postbiotic sample 1 is only in the inoculation strain, animal Bifidobacterium lactis LIHUO 01 and Lactobacillus plantarum P9 are inoculated into the fermentation medium at a ratio of 1:1, and the inoculation amount is 3% (v / v).

[0077] Preparation of postbiotic sample 6: The difference between the postbiotic sample 1 is only in the inoculation strain, animal Bifidobacterium lactis LIHUO 01 and Lactobacillus gasseri (Xi'an Yatu Biological Technology Co., Ltd.) are inoculated into the fermentation medium at a ratio of 1:1, and the inoculation amount is 3% (v / v).

[0078] Animal Bifidobacterium lactis LIHUO 01 postbiotic contains rich organic acids, including lactic acid, acetic acid and succinic acid, and contains a large amount of tryptophan metabolites, among which the content of indole-3-lactic acid in postbiotic sample 5 reaches 150.6 mg / kg. From the results of postbiotic sample 5, it can be seen that the total viable count and indole-3-lactic acid content of LIHUO 01 and Lactobacillus plantarum P9 after compounding are higher than those of postbiotic sample 1, which shows that strain compounding has a synergistic fermentation effect on the production of indole-3-lactic acid.

[0079] Table 3 Physicochemical indicators of animal Bifidobacterium lactis LIHUO 01 fermentation sample

[0080] Table 4 Physicochemical indicators of animal Bifidobacterium lactis LIHUO 01 postbiotic sample

[0081] Example 4 Relief effect of animal Bifidobacterium lactis LIHUO 01 postbiotic on loperamide-induced constipation in mice (1) Animal feeding and grouping: after adaptive feeding for 7 days, SD rats were randomly grouped according to body weight (normal group, model group, positive control group, postbiotic sample 1 group and postbiotic sample 5 group), 6 rats in each group.

[0082] (2) Modeling and administration: After the start of the experiment, the samples were intervened every day. The probiotic sample 1 and probiotic sample 5 groups were respectively given the prepared probiotic sample 1 and probiotic sample 5, and the daily gavage amount was 650 mg / kg body weight / day (equivalent to 7.2 g / day for humans). The positive drug group was given 1.80 g / kg polyethylene glycol 4000 dispersion by gavage every day. The normal group and the model group were given the same volume (1 mL) of PBS by gavage every day. On the 8th day of intervention, the model was injected with loperamide (10 mg / kg), 2 times a day, while the samples were intervened by gavage.

[0083] (3) Effect on the water content of animal feces: On the 14th day, the experimental rats were single-caged, and the defecation frequency within 1 hour was counted. The wet weight of feces was weighed, and the dry weight of feces was weighed after drying at 60°C for 24 hours. The water content of feces (%) = (m wet weight - m dry weight) / m wet weight x 100%. From Figure 1 It can be seen that after loperamide injection modeling, the water content of feces in the model group is significantly reduced, and the mice show high fecal hardness and difficulty in defecation. After intervention with positive drugs, probiotic sample 1 and probiotic sample 5, the water content of feces is significantly improved.

[0084] (4) Effect on intestinal propulsion rate of animals: After 24 hours of fasting without water feeding on the 14th day, the intestinal propulsion rate was measured. Each rat was given 2 mL of Evans blue paste (8 g of starch, 16 g of milk powder, 8 g of sugar, 5 g of carboxymethyl cellulose, and 7 mL of 1% Evans blue solution were dissolved in about 250 mL of distilled water, and then the mixture was mixed and made up to 300 mL of blue semi-solid paste) by gavage. After 15 minutes, the whole intestinal segment of the rat was dissected and stripped, placed on a glass flat plate, and the distance from the pylorus to the ileocecal junction and the distance from the pylorus to the Evans blue front were measured. The propulsion percentage was calculated using the formula: Intestinal propulsion rate% = distance from the blue front to the pylorus / total length of the small intestine x 100%. The intestinal propulsion rate of the model group was significantly lower than that of the normal group (P<0.05), indicating that the loperamide modeling was successful. Compared with the model group, probiotic sample 1 and probiotic sample 5 significantly improved the intestinal propulsion rate (P<0.05) Figure 2 ).

[0085] (5) Effect on the serum motilin (MTL) level of animals: The blood was taken from the orbital venous capillary of the animals, and the serum was separated. The MTL level in the sample was detected using a rat motilin ELISA detection kit (Jianglai Biological JL12355-96T). From Figure 3 It can be concluded that probiotic sample 1 and probiotic sample 5 can increase the MTL level in the serum of rats (P<0.05).

[0086] The above merely provides the preferred embodiment of the present application, and is not used to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A preparation method of a probiotic containing Bifidobacterium animalis lactis LIHUO 01, the Bifidobacterium animalis lactis LIHUO 01 has a preservation number of GDMCC No: 65512, characterized in that, The preparation method is that animal Bifidobacterium lactis LIHUO 01 is inoculated in a fermentation medium alone or in combination with probiotics to obtain a fermentation broth, and then the fermentation broth is subjected to heat inactivation to obtain a postbiotic containing animal Bifidobacterium lactis LIHUO 01. The fermentation medium comprises skimmed milk, soybean powder and water, and the mass ratio of the skimmed milk, the soybean powder and the water is (5-30):(1-25):(78-90). The probiotics are selected from one or more of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri and Lactobacillus gasseri.

2. The production method according to claim 1, characterized by, The mass ratio of the skimmed milk, the soybean powder and the water in the fermentation medium is 12:2:

86.

3. The preparation method according to claim 1, characterized in that, The fermentation medium further comprises a protease, and the addition amount of the protease is 10-1000 U / g of dry matter content.

4. The production method according to claim 3, characterized by, The protease is selected from one or more of papain, flavourzyme, acid protease, neutral protease, bromelain and alkaline protease.

5. The method of any one of claims 1-4, wherein, The postbiotic containing animal Bifidobacterium lactis LIHUO 01 comprises dead cells and / or bacterial lysates and / or strain metabolites, and the strain metabolites comprise indole-3-lactic acid, indole-3-acetic acid, indole-3-propionic acid, indole-3-formaldehyde, lactic acid, succinic acid and acetic acid.

6. The postbiotic containing animal Bifidobacterium lactis LIHUO 01 prepared by the preparation method according to any one of claims 1-5.

7. A fermentation medium, characterized in that, The fermentation medium comprises skimmed milk, soybean powder and water, and the mass ratio of the skimmed milk, the soybean powder and the water is (5-30):(1-25):(78-90).

8. Use of the postbiotic containing animal Bifidobacterium lactis LIHUO 01 according to claim 6 in the preparation of a product for relieving constipation.

9. A product for relief of constipation, characterized by, The product comprises the postbiotic containing animal Bifidobacterium lactis LIHUO 01 according to claim 6.

10. Use of the postbiotic containing animal Bifidobacterium lactis LIHUO 01 according to claim 6 in the preparation of a microecological preparation for producing indole-3-lactic acid, indole-3-acetic acid or short-chain fatty acids.

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