Intestine-derived probiotic composition as well as preparation method and application thereof

Abstract

The invention relates to the technical field of probiotics, in particular to an intestine-derived probiotic composition as well as a preparation method and application thereof. The lactobacillus reuteri and lactobacillus rhamnosus compound preparation specifically comprises the following components in parts by weight: 12-16 parts of lactobacillus reuteri, 11-15 parts of lactobacillus rhamnosus, 10-14 parts of bifidobacterium longum, 10-14 parts of lactobacillus acidophilus, 8-12 parts of lactobacillus, 8-12 parts of lactobacillus brevis, 5-9 parts of bacillus coagulans, 3-7 parts of lactobacillus gasseri and 14-18 parts of auxiliary materials. The probiotic composition provided by the invention can effectively improve intestinal injury caused by infection of harmful bacteria, and can enable an organism to gradually return to normal. Compared with the prior art, the probiotic composition provided by the invention has the advantages that the content of TNF-a (Tumor Necrosis Factor-a), IL-6 (Interleukin-6) and IL-10 (Interleukin-10) of proinflammatory cytokines is reduced, and the expression of ZO-1 and Occludin proteins is improved, so that the probiotic composition provided by the invention can be used for improving intestinal inflammation and improving intestinal barrier function damage caused by bacterial infection.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotics, and in particular to an intestinal probiotic composition and a preparation method and application thereof. Background Art

[0002] Probiotics are living microorganisms that, when ingested in sufficient quantities, produce health benefits to the host. According to the "List of Bacteria That Can Be Used in Food" and the "List of Bacteria That Can Be Used in Infant Food" issued by the National Health Administration, strains that may meet the definition of "probiotics" mainly come from the genera Lactobacillus and Bifidobacterium. As a resident intestinal flora, lactic acid bacteria can directly exert an antioxidant effect in the intestine, which is the key site of oxidative damage in the body. Moreover, after colonization in the intestine, lactic acid bacteria can continue to proliferate, produce more lactic acid bacteria, and continuously remove free radicals in the intestine. Therefore, lactic acid bacteria are becoming more and more popular as a probiotic.

[0003] Probiotics are a type of active microorganisms that are beneficial to the host by colonizing in the human body and changing the composition of the flora in a certain part of the host. By regulating the host mucosal and systemic immune function or by regulating the balance of intestinal flora, probiotics can promote nutrient absorption and maintain intestinal health, which is beneficial to human health. Therefore, the development of more comprehensive probiotic preparations can rebuild the human intestinal microecology and restore the homeostasis of intestinal flora.

[0004] In the probiotics preparation process, freeze drying (or lyophilization) is considered to be a suitable method to ensure the extended shelf life of most probiotic products. However, it is well known that this process also puts stress on the live bacteria. The freeze drying process is indeed a "challenge" to the viability of the probiotic strains. Therefore, in order to maintain an effective dose, most probiotic products are put into the production process with three to ten times more bacteria than the amount stated on the product label.

[0005] During the freeze-drying process, probiotics will be fatally damaged to a great extent, and when exposed to gastric acid and bile in the human body in this damaged state, the viability of probiotics will be further reduced due to fatal pressure. All of the above reasons have led to a significant reduction in the efficacy of probiotics. Therefore, how to improve the vitality of probiotic strains is also a technical problem that those skilled in the art continue to solve. Summary of the invention

[0006] The purpose of the present invention is to provide an intestinal probiotic composition and a preparation method and application thereof.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The invention provides an enteric probiotic composition, which is prepared from the following components in parts by weight: 12 to 16 parts of Lactobacillus reuteri, 11 to 15 parts of Lactobacillus rhamnosus, 10 to 14 parts of Bifidobacterium longum, 10 to 14 parts of Lactobacillus acidophilus, 8 to 12 parts of Lactobacillus, 8 to 12 parts of Lactobacillus brevis, 5 to 9 parts of Bacillus coagulans, 3 to 7 parts of Lactobacillus gasseri and 14 to 18 parts of auxiliary materials.

[0009] Preferably, the auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate.

[0010] Preferably, the mass ratio of the coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate is 13-17: 0.4-0.8: 0.9-1.3: 5-9: 5-9: 20-26.

[0011] Preferably, the bacterial content of the Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri is 500-700 million / g.

[0012] The present invention also provides a method for preparing the intestinal probiotic composition, comprising the following steps:

[0013] (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium for activation and culturing to obtain a seed solution;

[0014] (2) inoculating the seed solution obtained in step (1) into a culture medium for expansion and culture to obtain a bacterial solution;

[0015] (3) centrifuging and washing the bacterial solution obtained in step (2) to obtain bacterial cells;

[0016] (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri;

[0017] (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

[0018] Preferably, the culture medium in steps (1) and (2) uses water as solvent and comprises the following components: 15-19 g / L glucose, 4-6 g / L yeast extract, 9-11 g / L peptone, 0.5-1.5 g / L sodium chloride, 3-5 g / L potassium dihydrogen phosphate, 3-5 g / L potassium hydrogen phosphate, 0.5-1.5 g / L calcium chloride, 0.5-1.5 g / L magnesium sulfate heptahydrate, 4-6 g / L sodium acetate, and 4-6 g / L triammonium citrate.

[0019] Preferably, the inoculation ratio of the bacterial strain in step (1) is 2-4% (V / V); and the inoculation ratio of the seed liquid in step (2) is 3-5% (V / V).

[0020] Preferably, the activation culture and expansion culture in steps (1) and (2) are carried out at a temperature of 34 to 38° C. and for a time of 14 to 18 hours.

[0021] Preferably, the centrifugal speed in step (3) is 3500-4500 rpm, and the time is 8-12 min.

[0022] Preferably, the freeze-drying method in step (4) is: pre-freeze at -82 to -88°C for 3 to 4 hours, and then freeze-dry in a vacuum freeze dryer for 20 to 26 hours. The freeze-drying parameters are: vacuum degree 0.1Mpa to 0.2Mpa; cold trap temperature is -60°C to -65°C.

[0023] The present invention also provides the use of the intestinal probiotic composition in preparing a health product for improving intestinal health.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The intestinal probiotic composition provided by the present invention, wherein the added Lactobacillus rhamnosus has the effect of regulating intestinal hormones, and Bifidobacterium longum and Lactobacillus reuteri can significantly improve intestinal inflammation caused by bacterial infection and significantly improve the impaired intestinal barrier function caused by bacterial infection. The addition of Lactobacillus acidophilus can improve the body's ability to resist infection. Lactobacillus belongs to beneficial bacteria, which can help inhibit the reproduction of spoilage bacteria and pathogenic bacteria after entering the intestine, help maintain the balance of intestinal flora, and reduce the risk of intestinal infection. The addition of Bacillus coagulans and Lactobacillus gasseri can regulate the balance of intestinal flora.

[0026] The auxiliary materials used in the present invention play the role of probiotic activator, and can also prevent the oxidation of probiotics and prevent the functional loss of probiotics. Coenzyme Q10 can protect the integrity of the probiotic biofilm, and the addition of ornithine and tyrosine can activate freeze-dried probiotics, and is an active activator of probiotics. Sorbitol can balance the osmotic pressure, dextran and sodium citrate are stabilizers, and the above three components are used together to effectively avoid the influence of gastric juice stimulation or other types of stimulation on the activity of probiotics and the loss of probiotics.

[0027] The probiotic composition provided by the present invention can effectively improve intestinal damage caused by harmful bacteria infection, and can gradually restore the body to normal. It can also reduce the content of pro-inflammatory cytokines TNF-a, IL-6 and IL-10, and increase the expression of ZO-1 and Occludin proteins. Therefore, the probiotic composition provided by the present invention can improve intestinal inflammation and intestinal barrier function damage caused by bacterial infection. DETAILED DESCRIPTION

[0028] The present invention provides an intestinal probiotic composition, which is prepared from the following components in parts by weight: 12-16 parts of Lactobacillus reuteri, 11-15 parts of Lactobacillus rhamnosus, 10-14 parts of Bifidobacterium longum, 10-14 parts of Lactobacillus acidophilus, 8-12 parts of Lactobacillus acidophilus, 8-12 parts of Lactobacillus brevis, 5-9 parts of Bacillus coagulans, 3-7 parts of Lactobacillus gasseri and 14-18 parts of auxiliary materials; preferably 13-15 parts of Lactobacillus reuteri, 12-1 4 parts, Bifidobacterium longum 11-13 parts, Lactobacillus acidophilus 11-13 parts, Lactobacillus acidophilus 9-11 parts, Lactobacillus brevis 9-11 parts, Bacillus coagulans 6-8 parts, Lactobacillus gasseri 4-6 parts and 15-17 parts of auxiliary materials; further preferably 14 parts of Lactobacillus reuteri, 13 parts of Lactobacillus rhamnosus, 12 parts of Bifidobacterium longum, 12 parts of Lactobacillus acidophilus, 10 parts of Lactobacillus acidophilus, 10 parts of Lactobacillus brevis, 7 parts of Bacillus coagulans, 5 parts of Lactobacillus gasseri and 16 parts of auxiliary materials.

[0029] In the present invention, the auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate.

[0030] In the present invention, the mass ratio of coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate is 13-17: 0.4-0.8: 0.9-1.3: 5-9: 5-9: 20-26; preferably 14-16: 0.5-0.7: 1.0-1.2: 6-8: 6-8: 21-25; further preferably 15: 0.6: 1.1: 7: 7: 23.

[0031] In the present invention, the bacterial content of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri is 500 million to 700 million / g, preferably 600 million / g.

[0032] The present invention also provides a method for preparing the intestinal probiotic composition, comprising the following steps:

[0033] (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium for activation and culturing to obtain a seed solution;

[0034] (2) inoculating the seed solution obtained in step (1) into a culture medium for expansion and culturing to obtain a bacterial solution;

[0035] (3) centrifuging and washing the bacterial solution obtained in step (2) to obtain bacterial cells;

[0036] (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri;

[0037] (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

[0038] In the present invention, the culture medium in steps (1) and (2) uses water as a solvent and comprises the following components: 15-19 g / L glucose, 4-6 g / L yeast extract, 9-11 g / L peptone, 0.5-1.5 g / L sodium chloride, 3-5 g / L potassium dihydrogen phosphate, 3-5 g / L potassium hydrogen phosphate, 0.5-1.5 g / L calcium chloride, 0.5-1.5 g / L magnesium sulfate heptahydrate, 4-6 g / L sodium acetate, and 4-6 g / L triammonium citrate; preferably 16-18 g / L glucose, 5 g / L yeast extract, 10 g / L / L peptone, 0.7-1.3g / L sodium chloride, 4g / L potassium dihydrogen phosphate, 4g / L potassium hydrogen phosphate, 0.7-1.3g / L calcium chloride, 0.7-1.3g / L magnesium sulfate heptahydrate, 5g / L sodium acetate, 5g / L triammonium citrate; further preferably 17g / L glucose, 5g / L yeast extract, 10g / L peptone, 1g / L sodium chloride, 4g / L potassium dihydrogen phosphate, 4g / L potassium hydrogen phosphate, 1g / L calcium chloride, 1g / L magnesium sulfate heptahydrate, 5g / L sodium acetate, 5g / L triammonium citrate.

[0039] In the present invention, the inoculation ratio of the bacterial strain in step (1) is 2-4% (V / V), preferably 3% (V / V).

[0040] In the present invention, the inoculation ratio of the seed solution in step (2) is 3-5% (V / V); preferably 4% (V / V).

[0041] In the present invention, the temperature of the activation culture and the expansion culture in steps (1) and (2) is 34-38°C, and the time is 14-18h; preferably, the temperature is 35-37°C, and the time is 15-17h; further preferably, the temperature is 36°C, and the time is 16h.

[0042] In the present invention, the centrifugal speed in step (3) is 3500-4500 rpm, and the time is 8-12 min; preferably, the centrifugal speed is 3700-4300 rpm, and the time is 9-11 min; further preferably, the centrifugal speed is 3900-4100 rpm, and the time is 10 min; more preferably, the centrifugal speed is 4000 rpm, and the time is 10 min.

[0043] In the present invention, the freeze-drying method in step (4) is: pre-freezing at -82 to -88°C for 3 to 4 hours, and then freeze-drying in a vacuum freeze dryer for 20 to 26 hours, and the freeze-drying parameters are: vacuum degree 0.1Mpa to 0.2Mpa; cold trap temperature is -60°C to -65°C; preferably, the freeze-drying method is: pre-freezing at -83 to -87°C for 3.5 hours, and then freeze-drying in a vacuum freeze dryer for 21 to 25 hours, and the freeze-drying parameters are: vacuum degree 0.2Mpa; cold trap temperature is -61°C to -64°C; further preferably, the freeze-drying method is: pre-freezing at -85°C for 3.5 hours, and then freeze-drying in a vacuum freeze dryer for 23 hours, and the freeze-drying parameters are: vacuum degree 0.2Mpa; cold trap temperature is -63°C.

[0044] The present invention also provides the use of the intestinal probiotic composition in preparing a health product for improving intestinal health.

[0045] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0046] Example 1

[0047] A gut-derived probiotic composition is prepared from the following components in parts by mass: 12 parts of Lactobacillus reuteri, 11 parts of Lactobacillus rhamnosus, 10 parts of Bifidobacterium longum, 10 parts of Lactobacillus acidophilus, 8 parts of Lactobacillus, 8 parts of Lactobacillus brevis, 5 parts of Bacillus coagulans, 3 parts of Lactobacillus gasseri and 14 parts of auxiliary materials.

[0048] The auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate, and the mass ratio is 13:0.4:0.9:5:5:20.

[0049] The bacterial contents of the Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri are all 500 million / g.

[0050] The preparation method of the intestinal probiotic composition comprises the following steps:

[0051] (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium at an inoculation ratio of 2% (V / V) and culturing at 34° C. for 14 hours to obtain a seed solution;

[0052] (2) inoculating the seed solution obtained in step (1) into a culture medium at an inoculation ratio of 3% (V / V) and culture at 34° C. for 14 h to obtain a bacterial solution;

[0053] (3) centrifuging the bacterial solution obtained in step (2) at 3500 rpm for 8 min and washing to obtain bacterial cells;

[0054] (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidobacillus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri; the freeze-drying method is: pre-freezing at -82°C for 3h, and then freeze-drying in a vacuum freeze dryer for 20h, and the freeze-drying parameters are: vacuum degree 0.1Mpa; cold trap temperature is -60°C.

[0055] (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

[0056] The culture medium in steps (1) and (2) uses water as a solvent and comprises the following components: 15 g / L glucose, 4 g / L yeast extract powder, 9 g / L peptone, 0.5 g / L sodium chloride, 3 g / L potassium dihydrogen phosphate, 0.5 g / L potassium hydrogen phosphate, 0.5 g / L calcium chloride, 0.5 g / L magnesium sulfate heptahydrate, 4 g / L sodium acetate, and 4 g / L triammonium citrate.

[0057] Example 2

[0058] A gut-derived probiotic composition is prepared from the following components in parts by weight: 16 parts of Lactobacillus reuteri, 15 parts of Lactobacillus rhamnosus, 14 parts of Bifidobacterium longum, 14 parts of Lactobacillus acidophilus, 12 parts of Lactobacillus, 12 parts of Lactobacillus brevis, 9 parts of Bacillus coagulans, 7 parts of Lactobacillus gasseri and 18 parts of auxiliary materials.

[0059] The auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate, and the mass ratio is 17:0.8:1.3:9:9:26.

[0060] The bacterial contents of the Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri are all 700 million / g.

[0061] The preparation method of the intestinal probiotic composition comprises the following steps:

[0062] (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium at an inoculation ratio of 4% (V / V) for activation culture at 38° C. for 18 hours to obtain a seed solution;

[0063] (2) inoculating the seed solution obtained in step (1) into a culture medium at an inoculation ratio of 5% (V / V) and culture at 38° C. for 18 h to obtain a bacterial solution;

[0064] (3) centrifuging the bacterial solution obtained in step (2) at 4500 rpm for 12 min and washing to obtain bacterial cells;

[0065] (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidobacillus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri; the freeze-drying method is: pre-freezing at -88°C for 4 hours, and then freeze-drying in a vacuum freeze dryer for 26 hours, and the freeze-drying parameters are: vacuum degree 0.2 MPa; cold trap temperature is -65°C.

[0066] (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

[0067] The culture medium in steps (1) and (2) uses water as a solvent and comprises the following components: 19 g / L glucose, 6 g / L yeast extract powder, 11 g / L peptone, 1.5 g / L sodium chloride, 5 g / L potassium dihydrogen phosphate, 5 g / L potassium hydrogen phosphate, 1.5 g / L calcium chloride, 1.5 g / L magnesium sulfate heptahydrate, 6 g / L sodium acetate, and 6 g / L triammonium citrate.

[0068] Example 3

[0069] A gut-derived probiotic composition is prepared from the following components in parts by weight: 14 parts of Lactobacillus reuteri, 13 parts of Lactobacillus rhamnosus, 12 parts of Bifidobacterium longum, 12 parts of Lactobacillus acidophilus, 10 parts of Lactobacillus, 10 parts of Lactobacillus brevis, 7 parts of Bacillus coagulans, 5 parts of Lactobacillus gasseri and 16 parts of auxiliary materials.

[0070] The auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate, and the mass ratio is 15:0.6:1.1:7:7:23.

[0071] The bacterial contents of the Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri are all 600 million / g.

[0072] The preparation method of the intestinal probiotic composition comprises the following steps:

[0073] (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium at an inoculation ratio of 3% (V / V) and culturing at 36° C. for 16 hours to obtain a seed solution;

[0074] (2) inoculating the seed solution obtained in step (1) into a culture medium at an inoculation ratio of 4% (V / V) and culture at 36° C. for 16 h to obtain a bacterial solution;

[0075] (3) centrifuging the bacterial solution obtained in step (2) at 4000 rpm for 10 min and washing to obtain bacterial cells;

[0076] (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri; the freeze-drying method is: pre-freezing at -85°C for 3.5 hours, and then freeze-drying in a vacuum freeze dryer for 23 hours, and the freeze-drying parameters are: vacuum degree 0.2 MPa; cold trap temperature is -63°C.

[0077] (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

[0078] The culture medium in steps (1) and (2) uses water as a solvent and comprises the following components: 17 g / L glucose, 5 g / L yeast extract powder, 10 g / L peptone, 1 g / L sodium chloride, 4 g / L potassium dihydrogen phosphate, 4 g / L potassium hydrogen phosphate, 1 g / L calcium chloride, 1 g / L magnesium sulfate heptahydrate, 5 g / L sodium acetate, and 5 g / L triammonium citrate.

[0079] Experimental Example 1

[0080] 80 Balb / c mice (Alliance Biotechnology Co., Ltd.), 21 days old, were randomly divided into 8 groups after 3 days of pre-feeding.

[0081] Blank control group: no special treatment;

[0082] Model group: Escherichia coli suspension (1*10 8 CFU / mL) 1mL gavage;

[0083] The product prepared by the method of Example 3 was used as the experimental group;

[0084] In control group 1, other methods were the same as those in Example 3, except that Lactobacillus brevis and Bacillus coagulans were replaced by an equal amount of Lactobacillus reuteri.

[0085] Control group 2: other methods are the same as those in Example 3, except that no auxiliary materials are used.

[0086] Control group 3: other methods were the same as those in Example 3, except that coenzyme Q10 and ornithine were replaced with an equal amount of sodium citrate in the auxiliary materials.

[0087] Control group 4, other methods are the same as those in Example 3, except that freeze-drying is performed with reference to the prior art method (CN201910516435.4), and the specific method is: refrigerate at 4°C for 30 min, -20°C for 1 h, and -80°C for 1 h. After cooling in steps, the solid mixture is spread in a freeze dryer for 14 h to obtain freeze-dried powder.

[0088] Prior art group: The probiotic composition provided by CN201910909883.0, the probiotic concentration is 1*10 10 CFU / mL.

[0089] Example 3, control groups 1 to 4, and the prior art groups were all gavaged with E. coli according to the method of the model group, and the corresponding product (gavage after dissolving in water) was gavaged at a dose of 10 g / kg, once a day for 3 days.

[0090] After the experiment was completed, the weight changes of mice were investigated, blood was collected from the eyeballs of mice, serum was prepared, and cytokines TNF-a, IL-6 and IL-10 in serum were detected using enzyme-linked immunosorbent assay kits. The specific experimental steps were carried out according to the instructions of the kits, and the protein expression levels of tight junction proteins ZO-1 and occludin in the mouse colon were detected using immunological assay methods Western blot. The survey results were averaged, and the results are shown in Tables 1 and 2.

[0091] Table 1 Body weight changes

[0092] Weight change rate % Blank control group +10.3％ Model Group -25.0％ Experimental Group +7.8％ Control group 1 +4.0％ Control group 2 +3.3％ Control group 3 +5.6％ Control group 4 +4.2％ Existing Technology Group +5.3％

[0093] "+" represents weight gain and "-" represents weight loss

[0094] Table 2 Changes of cytokines (unit: pg / mL)

[0095] TNF-a IL-6 IL-10 Blank control group 226.4 187.9 436.1 Model Group 1236.7 2155.6 1768.4 Experimental Group 362.5 484.7 703.5 Control group 1 556.0 684.1 812.6 Control group 2 722.8 1364.4 1225.3 Control group 3 468.4 788.2 1012.5 Control group 4 619.3 822.4 1186.7 Existing Technology Group 576.6 741.9 912.1

[0096] Table 3 Protein expression

[0097]

[0098]

[0099] It can be seen from the contents recorded in Tables 1 to 3 that the probiotic composition provided by the present invention has excellent technical effects of improving intestinal inflammation caused by Escherichia coli infection and improving intestinal barrier, and its use effect is also significantly better than the related products of the prior art.

[0100] From the data comparison between the experimental group, the blank control group and the model group, it can be seen that the weight of the mice in the model group decreased significantly, and the weight of the mice in the experimental group increased significantly, which shows that after the use of probiotics, the body of the mice gradually returned to normal. The levels of proinflammatory cytokines TNF-a, IL-6 and IL-10 in the serum of the mice in the model group were significantly increased, and after the probiotics intervention, the cytokine levels were greatly reduced, which shows that the probiotics provided by the present invention can improve intestinal inflammation. The expression of intestinal tight junction proteins ZO-1 and Occludin in the model group mice was reduced, and after the probiotics intervention, the expression of proteins ZO-1 and Occludin was greatly increased, which shows that the use of the probiotics of the present invention can improve the intestinal barrier function damage of mice infected with Escherichia coli.

[0101] It can be seen from the data of control groups 1 to 3 that only by using the complete formulation of the present invention can the best use effect be achieved. If the complete strain of the present invention is not used, or if the auxiliary materials are not added, or the auxiliary materials are incomplete, the efficacy of the composition will be significantly affected.

[0102] It can also be seen from the data of control group 4 that only the freeze-drying method of the present invention can achieve the best use effect. If the method of the prior art is used, the activity of the probiotics will be affected, which will in turn affect the use effect of the probiotics.

[0103] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A gut-derived probiotic composition, characterized in that: The invention is prepared from the following components in parts by weight: 12 to 16 parts of Lactobacillus reuteri, 11 to 15 parts of Lactobacillus rhamnosus, 10 to 14 parts of Bifidobacterium longum, 10 to 14 parts of Lactobacillus acidophilus, 8 to 12 parts of Lactobacillus acidophilus, 8 to 12 parts of Lactobacillus brevis, 5 to 9 parts of Bacillus coagulans, 3 to 7 parts of Lactobacillus gasseri and 14 to 18 parts of auxiliary materials.

2. The intestinal-derived probiotic composition according to claim 1, characterized in that: The auxiliary materials are: coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate.

3. The intestinal-derived probiotic composition according to claim 2, characterized in that: The mass ratio of the coenzyme Q10, ornithine, tyrosine, sorbitol, dextran and sodium citrate is 13-17: 0.4-0.8: 0.9-1.3: 5-9: 5-9: 20-26.

4. The intestinal-derived probiotic composition according to claim 1, characterized in that: The bacterial contents of the Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri are all 500 million to 700 million per gram.

5. The method for preparing the intestinal probiotic composition according to any one of claims 1 to 4, characterized in that: The steps include: (1) inoculating Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri into a culture medium for activation and culturing to obtain a seed solution; (2) inoculating the seed solution obtained in step (1) into a culture medium for expansion and culturing to obtain a bacterial solution; (3) centrifuging and washing the bacterial solution obtained in step (2) to obtain bacterial cells; (4) freeze-drying the bacterial cells obtained in step (3) to obtain freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri; (5) The freeze-dried powders of Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus brevis, Bacillus coagulans and Lactobacillus gasseri obtained in step (4) are mixed with auxiliary materials to obtain an enterogenic probiotic composition.

6. The preparation method according to claim 5, characterized in that: The culture medium in steps (1) and (2) uses water as a solvent and comprises the following components: 15-19 g / L glucose, 4-6 g / L yeast extract powder, 9-11 g / L peptone, 0.5-1.5 g / L sodium chloride, 3-5 g / L potassium dihydrogen phosphate, 3-5 g / L potassium hydrogen phosphate, 0.5-1.5 g / L calcium chloride, 0.5-1.5 g / L magnesium sulfate heptahydrate, 4-6 g / L sodium acetate, and 4-6 g / L triammonium citrate.

7. The preparation method according to claim 5, characterized in that: The inoculation ratio of the bacterial strain in step (1) is 2-4% (V / V); the inoculation ratio of the seed liquid in step (2) is 3-5% (V / V).

8. The preparation method according to claim 5, characterized in that: The activation culture and expansion culture in steps (1) and (2) are both carried out at a temperature of 34 to 38° C. and for a time of 14 to 18 hours.

9. The preparation method according to claim 5, characterized in that: The centrifugal speed in step (3) is 3500-4500 rpm, and the time is 8-12 min. The freeze-drying method in step (4) is: pre-freeze at -82-88°C for 3-4 h, and then freeze-dry in a vacuum freeze dryer for 20-26 h. The freeze-drying parameters are: vacuum degree 0.1Mpa-0.2Mpa; cold trap temperature -60°C-65°C.

10. Use of the intestinal probiotic composition according to any one of claims 1 to 4 in the preparation of a health product for improving intestinal health.

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