Probiotic composition and application

Abstract

The invention belongs to the technical field of food microorganisms, and particularly relates to a probiotic composition and application. The feed additive comprises the following raw materials in parts by weight: 1-5 parts of a probiotic mixture, 10-20 parts of a prebiotic mixture and 1-3 parts of an accelerant, the probiotic mixture comprises lactococcus lactis and phytobacterium plantarum, and the mass ratio of the lactococcus lactis to the phytobacterium plantarum is 1: (0.5-1); the prebiotic mixture is prepared from oligosaccharide, polyphenol and protein hydrolysate, and the mass ratio of the oligosaccharide to the polyphenol to the protein hydrolysate is 1: (0.3-0.5): (0.5-1); the accelerant is prepared from plant polysaccharide and chlorogenic acid, and the mass ratio of the plant polysaccharide to the chlorogenic acid is 1: (0.01-0.05). According to the composition disclosed by the invention, the probiotics and the prebiotics in a specific proportion are taken as effective active components, and the probiotic composition formed by combining the probiotics and the prebiotics can be used for synergistically regulating the intestinal flora structure of a human body, and has a remarkable effect in the aspect of preventing or treating diarrhea or intestinal flora disorder and the like; health food, health care products or medicines prepared from the compound have potential application value.

Description

Technical Field

[0001] This invention belongs to the field of food microbiology technology, and more specifically, relates to a probiotic composition and its uses. Background Technology

[0002] The human gut microbiota is easily disrupted by a multitude of factors, including environmental factors, diet and lifestyle habits, psychological factors, disease states, cancer treatment, antibiotic use, and age. This is known as gut microbiota dysbiosis, which can manifest as a deficiency of beneficial bacteria, an overgrowth of pathogens, impaired intestinal barrier function, and intestinal inflammation. This can further lead to gastrointestinal diseases in the host, including constipation, diarrhea, abdominal pain, and bloating. In severe cases, it can develop into inflammatory bowel disease, ulcerative colitis, irritable bowel syndrome, and other diseases, greatly affecting human health and quality of life.

[0003] Given the close relationship between the gut microbiota and human health, consuming fermented milk has a positive impact on the composition of the gut microbiota and the health of the host. Different strains of bacteria, raw materials, raw material ratios, and fermentation processes affect fermentation metabolites, thereby producing different health benefits.

[0004] Lactococcus lactis subsp. lactis WG-lla 26 is a proprietary product of the applicant. Preliminary studies have shown that this strain possesses superior precursor utilization and the ability to produce specific flavor compounds, exhibiting a characteristic malty flavor in fermented milk products. Currently, no functional products containing this strain have been disclosed.

[0005] Therefore, there is a desire for a probiotic composition containing Lactococcus lactis subsp. lactis that will have a significant effect on improving gut health. Summary of the Invention

[0006] The purpose of this invention is to provide a probiotic composition and its use, which can effectively improve the intestinal flora and thus promote human health.

[0007] Therefore, the present invention provides the following technical solution.

[0008] The first aspect of the present invention provides a probiotic composition comprising, by weight, 1 to 5 parts of a probiotic mixture, 10 to 20 parts of a prebiotic mixture, and 1 to 3 parts of a promoter.

[0009] The probiotic mixture comprises Lactococcus lactis and Lactobacillus plantarum, wherein the mass ratio of Lactococcus lactis subsp. lactis to Lactobacillus plantarum is 1:(0.5-1).

[0010] The prebiotic mixture comprises oligosaccharides, polyphenols, and protein hydrolysates, wherein the mass ratio of oligosaccharides, polyphenols, and protein hydrolysates is 1:(0.3-0.5):(0.5-1).

[0011] The promoter comprises plant polysaccharides and chlorogenic acid, wherein the mass ratio of plant polysaccharides to chlorogenic acid is 1:(0.01-0.05).

[0012] In a preferred embodiment of the present invention, the lactococcus is Lactococcus lactis subsp. lactis WG-lla1a 26, with a viable count of 1×10⁻⁶. 8 ~1×10 10 CFU / g.

[0013] In a preferred embodiment of the present invention, the *Lactobacillus plantarum* is *Lactobacillus plantarum* WG-lp40 with a viable count of 1×10⁻⁶. 8 ~1×10 10 CFU / g.

[0014] In a preferred embodiment of the present invention, the oligosaccharide is selected from one or more of fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, xylooligosaccharides, lactulose, and human milk oligosaccharides (HMOs).

[0015] In a preferred embodiment of the present invention, the polyphenol is selected from one or more of curcumin, tea polyphenols, catechins, anthocyanins, and quercetin, or a mixture thereof.

[0016] In a preferred embodiment of the present invention, the protein hydrolysate is quinoa protein peptide and / or walnut polypeptide.

[0017] In a preferred embodiment of the present invention, the plant polysaccharide is selected from one or more of the following: red bean grass polysaccharide, black tea polysaccharide, ginseng polysaccharide, wolfberry polysaccharide, astragalus polysaccharide, ginseng polysaccharide, yam polysaccharide, rose polysaccharide, and inulin.

[0018] A second aspect of the invention provides the use of the probiotic composition as described above in the preparation of a medicine, food or health product for improving gut health.

[0019] A third aspect of the present invention provides a medicine, food, or health product comprising the probiotic composition as described above.

[0020] In a preferred embodiment of the present invention, the food is selected from at least one of dairy products, cereal products, soy products, condiments, ice cream, and beverages.

[0021] In a preferred embodiment of the present invention, the dairy product is selected from at least one of sterilized milk, reconstituted milk, fermented milk, yogurt, milk powder, formula milk powder, condensed milk, cheese, casein, and whey powder.

[0022] By employing the above technical solution, the present invention has at least the following advantages:

[0023] The composition of the present invention uses probiotics and prebiotics in a specific ratio as effective active ingredients. The probiotic composition formed by the combination of the two can synergistically regulate the structure of human intestinal flora and has significant effects in preventing or treating diarrhea or intestinal flora disorders. It has potential application value in the preparation of health foods, health care products or medicines.

[0024] The probiotic mixture of this invention comprises *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 and *Lactobacillus plantarum* WG-lp40, both unique to the applicant. *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 exhibits superior precursor utilization and the ability to produce specific flavor compounds, resulting in a characteristic malty flavor in fermented milk products. *Lactobacillus plantarum* WG-lp40 possesses a strong ability to metabolize phenyllactic acid and related metabolites, thus regulating the microecological balance and inhibiting the growth of pathogenic microorganisms. The combined use of these two active bacteria can effectively regulate human intestinal health, helping to prevent or treat problems such as diarrhea or intestinal flora imbalance.

[0025] The prebiotic of this invention is composed of oligosaccharides, polyphenols, and protein hydrolysates. The addition of prebiotics can not only promote and enhance the proliferation and activity of Lactococcus lactis and Lactobacillus plantarum, but also promote the proliferation of beneficial intestinal bacteria such as Lactobacillus and Bifidobacterium, while inhibiting the growth and activity of opportunistic pathogens such as fungi, Escherichia coli, and Enterococcus.

[0026] The promoter of this invention primarily comprises plant polysaccharides and chlorogenic acid. Among them, plant polysaccharides, such as red bean grass polysaccharide, possess various activities including immunomodulatory, antioxidant, and gut health-promoting effects. Studies have shown that plant polysaccharides can regulate intestinal immunity, not only increasing the diversity of gut microbiota, regulating its structure and composition, and restoring gut microbial balance, but also improving intestinal morphology, upregulating small intestinal tight junction proteins, and reducing intestinal permeability to maintain intestinal barrier integrity and ensure normal barrier function. Chlorogenic acid has antioxidant and anti-inflammatory effects. The promoter composed of these two substances not only synergistically promotes gut health but also provides a nutrient source for probiotics, ensuring their biological activity.

[0027] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below. Detailed Implementation

[0028] To make the technical means, creative features, achieved objectives, and effects of this invention readily understandable, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0029] Lactococcus lactis subsp. lactis WG-lla 26 was selected by the applicant through screening based on 3-methylbutyraldehyde, 3-methylbutanol, and 2,3-butanedione as characteristic flavor indicators, and named Lactococcus lactis subsp. lactis WG-lla 26. It was deposited on November 8, 2022, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 26090. Specific information can be found in application number CN202310569061.9, the full text of which is incorporated herein by reference. After initial screening based on fermentation time, fermentation temperature, passage stability, and post-acidification, and subsequent screening based on sensory analysis, differences in flavor substance metabolism, and precursor utilization ability, this strain showed high precursor utilization ability and characteristic flavor substance metabolism ability. Its fermentation in dairy products can produce a malty aroma characteristic flavor.

[0030] Lactiplantibacillus plantarum WG-lp40 was obtained by the applicant through screening in Tibetan milk curds and was deposited on August 28, 2023, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 28272. Specific information can be found in application number CN202411515333.8, the full text of which is incorporated herein by reference. Previous studies have shown that this strain has a strong ability to metabolize phenyllactic acid and related metabolites, can adjust the microecological balance, and inhibit the growth of pathogenic microorganisms. By applying this strain to the preparation process of mold-inhibiting flavored fermented milk and optimizing the original formula and preparation method, a flavored fermented milk with improved phenyllactic acid metabolism and its preparation method were obtained. The resulting mold-inhibiting flavored fermented milk can inhibit mold growth without the addition of antibacterial additives, reducing the risk of product spoilage caused by mold through the antibacterial properties of the probiotics themselves.

[0031] Example 1: Composition I and its preparation

[0032] Composition I, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The mass ratio of CFU / g is 1:0.75. The mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75. The mass ratio of red bean grass polysaccharide and chlorogenic acid in the promoter is 1:0.03.

[0033] Preparation of Composition I: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0034] Example 2: Composition II and its preparation

[0035] Composition II, by weight, comprises: 1 part probiotic mixture, 20 parts prebiotic mixture, and 1 part promoter; wherein the probiotic mixture contains Lactococcus lactis subsp. lactis WG-lla 26 (live count 1×10⁻⁶). 8 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 10 The mass ratio of CFU / g is 1:0.5, the mass ratio of galactooligosaccharides, anthocyanins and quinoa protein peptides in the prebiotic mixture is 1:0.3:1, and the mass ratio of wolfberry polysaccharide and chlorogenic acid in the promoter is 1:0.01.

[0036] Preparation of Composition II: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0037] Example 3: Composition III and its preparation

[0038] Composition III, by weight, comprises: 5 parts of a probiotic mixture, 10 parts of a prebiotic mixture, and 3 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (with a live count of 1 × 10⁻⁶). 10 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 8 The CFU / g mass ratio is 1:1, the mass ratio of xylooligosaccharide, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.5:0.5, and the mass ratio of rose polysaccharide and chlorogenic acid in the promoter is 1:0.05.

[0039] Preparation of Composition III: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0040] Example 4: Composition IV and its preparation

[0041] Composition IV, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The mass ratio of CFU / g is 1:0.75. The mass ratio of human milk oligosaccharide (HMO), tea polyphenols, and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75. The mass ratio of inulin and chlorogenic acid in the promoter is 1:0.03.

[0042] Preparation of Composition IV: The above raw materials are thoroughly mixed in proportion to obtain the final product.

[0043] Comparative Example 1: Composition V and its preparation

[0044] Composition V, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture is *Lactobacillus plantarum* WG-lp40 (live count 1×10⁻⁶). 9 The mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75, and the mass ratio of red bean grass polysaccharide and chlorogenic acid in the promoter is 1:0.03.

[0045] Preparation of composition V: The above raw materials are thoroughly mixed in proportion to obtain the final product.

[0046] Comparative Example 2: Composition VI and its preparation

[0047] Composition VI, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture is *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (live count 1×10⁻⁶). 9 The mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75, and the mass ratio of red bean grass polysaccharide and chlorogenic acid in the promoter is 1:0.03.

[0048] Preparation of Composition VI: Mix the above raw materials thoroughly according to the specified proportions to obtain the final product.

[0049] Comparative Example 3: Composition VII and its preparation

[0050] Composition VII, by weight, comprises: 15 parts of a prebiotic mixture and 2 parts of a promoter; wherein the mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75, and the mass ratio of red bean grass polysaccharide and chlorogenic acid in the promoter is 1:0.03.

[0051] Preparation of Composition VII: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0052] Comparative Example 4: Composition VIII and its preparation

[0053] Composition VIII, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-11a 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The CFU / g mass ratio is 1:0.75, the mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75, and the promoter is red bean grass polysaccharide.

[0054] Preparation of Composition VIII: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0055] Comparative Example 5: Composition IX and its preparation

[0056] Composition IX, by weight, comprises: 3 parts of a probiotic mixture, 15 parts of a prebiotic mixture, and 2 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The CFU / g mass ratio is 1:0.75, the mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75, and the promoter is chlorogenic acid.

[0057] Preparation of Composition IX: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0058] Comparative Example 6: Composition X and its preparation

[0059] Composition X, by weight, comprises: 3 parts of a probiotic mixture and 15 parts of a prebiotic mixture; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-lla 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The CFU / g mass ratio is 1:0.75, and the mass ratio of fructooligosaccharides, tea polyphenols and quinoa protein peptides in the prebiotic mixture is 1:0.4:0.75.

[0060] Preparation of composition X: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0061] Comparative Example 7: Composition XI and its preparation

[0062] Composition XI, by weight, comprises: 3 parts of a probiotic mixture and 2 parts of a promoter; wherein the probiotic mixture contains *Lactococcus lactis* subsp. *lactococcus* WG-11a 26 (with a live count of 1 × 10⁻⁶). 9 CFU / g) and Lactobacillus plantarum WG-lp40 (live count 1×10⁻⁶) 9 The mass ratio of CFU / g is 1:0.75, and the mass ratio of red bean grass polysaccharide and chlorogenic acid in the accelerator is 1:0.03.

[0063] Preparation of composition XI: Mix the above raw materials thoroughly according to the proportions to obtain the final product.

[0064] Test 1: In vitro antibacterial test of the composition

[0065] Sample preparation: The compositions of Examples 1-4 and Comparative Examples 1-7 were dissolved in PBS buffer at a mass ratio of 1:100 to obtain sample solutions prepared from the compositions of Examples 1-4 and Comparative Examples 1-7, which were stored at 4°C for later use.

[0066] Four common pathogenic bacteria, namely Escherichia coli, Salmonella, Bacillus subtilis, and Staphylococcus aureus, were selected. The inhibitory effects of the compositions obtained in Examples 1-4 and Comparative Examples 1-7 on each pathogenic bacterium were determined using the agar diffusion method. Specifically, the following steps were taken: First, Escherichia coli, Salmonella, Bacillus subtilis, and Staphylococcus aureus were activated and inoculated into LB liquid medium at a ratio of 3%, and cultured at 37°C for 24 hours. After the culture was completed, 100 μL of the pathogenic bacterial suspension (concentration of 10%) was collected. 9 A plate containing cfu / mL was evenly spread onto LB solid medium. After spreading, the plate was checked until no visible water droplets remained. Three wells were evenly punched in each well. 200 μL of the sample solution from Examples 1-4 and Comparative Examples 1-7 was then pipetted into each well. The plate was placed in an incubator at 18°C ​​for 12 hours to diffuse, then transferred to an incubator at 37°C. The diameter of the inhibition zone was measured once a clear inhibition zone appeared. Each well was measured three times. The results are shown in Table 1.

[0067] Table 1 Summary of the antibacterial effects of each composition

[0068]

[0069]

[0070] As can be seen from the results in Table 1, the compositions of Examples 1-4 of the present invention have a more obvious antibacterial effect compared with the compositions of Comparative Examples 1-7.

[0071] Experiment 2: A granule containing a composition

[0072] 1. Granules containing the composition and their preparation

[0073] This experiment provides a tablet containing the composition, which is prepared by taking 25 parts of the composition, 10 parts of magnesium stearate, 10 parts of silicon dioxide and 5 parts of edible flavoring; mixing the above raw materials thoroughly, making granules, and packaging them into bags of 2g each.

[0074] Granules containing the compositions of Example 1 and Comparative Examples 1-7 were prepared according to the above method.

[0075] 2. Application of granules in relieving diarrhea in mice

[0076] Using BALB / c mice as the research subjects, different granular solutions were administered to the mice by gavage to detect the effects of these samples on the body weight, fecal water content, and serum immune inflammatory factors in the mice with diarrhea, in order to determine the efficacy of these samples in relieving bacterial diarrhea.

[0077] 2.1 Feed and Animal Sources

[0078] The basic feed (composed of the following percentages by weight: 20% wheat flour, 10% rice flour, 20% corn, 26% hulled wheat bran, 20% soybean, 2% fish meal, and 2% bone meal) was purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.

[0079] SPF-grade BALB / c mice (half male and half female, 6 weeks old, weighing 18-20g) were purchased from the Comparative Medicine Center of Yangzhou University. The experimental animals were housed in the experimental animal facility of the College of Food Science and Engineering, Yangzhou University. Laboratory temperature: 23.0±1.0℃, humidity: 50±5%, lighting conditions: cycled every 12 hours (8:00~20:00), with free access to water and food. All experimental conditions met the "Standards for Laboratory Animal Environment and Facilities" issued by the Ministry of Health of the People's Republic of China.

[0080] 2.2 Methods

[0081] The experimental mice were first acclimatized for one week, and then randomly divided into 11 groups using a random number table. Except for the control group (NC) of 5 mice, the other model groups each had 5 mice. The formal experimental period was 21 days. From days 1 to 7, the mice were acclimatized. From days 8 to 14, a diarrhea model was established using senna leaf-induced diarrhea: each mouse in the model group was administered 4 g / (kg·d) of senna leaf via gavage for 7 consecutive days to establish the diarrhea model. From days 9 to 18, the model group mice were randomly divided into 10 groups using a random number table: Example 1 group, Comparative Examples 1-7 groups, and the model group, with 5 mice in each group. There was no significant difference in body weight among the groups after grouping, indicating that the grouping was reasonable. The mice in each group were then treated as follows:

[0082] Example 1 group: Each mouse was administered 30 mg of the granules from Example 1 by gavage + 0.2 mL of sterile saline twice daily;

[0083] Comparative Examples 1-7: Same as Example 1, except that the granules were replaced with the corresponding granules of the corresponding examples;

[0084] Blank control group and model group: Each mouse was administered 0.2 ml of sterile saline by gavage twice a day.

[0085] During the experiment, the mice had free access to food and water. All groups of mice were fed standard feed. The enclosure was a constant temperature and humidity environment with a temperature of 25±2℃ and a humidity of 50%±5%. The lighting conditions were 12 hours of light and 12 hours of darkness.

[0086] 2.3 Detection Indicators

[0087] 2.3.1 Fecal Moisture Content: One day before the mice were sacrificed, as much feces as possible were collected, freeze-dried, and the moisture content before and after freezing was measured.

[0088] Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%;

[0089] 2.3.2 Determination of serum inflammatory factors TNF-α, IL-6, IL-10 and IgA: 24 hours after the last gavage, blood samples were collected from the orbital venous plexus of mice in each group. The samples were centrifuged at 4000g for 10 min, and the serum was collected and stored at -80℃. The expression levels of each indicator in the serum were determined by enzyme-linked immunosorbent assay.

[0090] 2.4 Experimental results: as shown in Table 2.

[0091] Table 2. Effects of different granule formulations on diarrheal mice.

[0092]

[0093]

[0094] As can be seen from the results in Table 2, compared with the granule products of Comparative Examples 1-7, the granule of Example 1 of the present invention has a more obvious effect in inhibiting diarrhea in mice, and can also significantly control the content of intestinal inflammatory factors in mice and increase antibody content.

[0095] Experiment 3: A dairy product containing the composition

[0096] 1. Dairy products and their preparation

[0097] This experiment provides a dairy product containing the composition, which, by weight percentage, consists of: 5% composition, 0.5% steviol glycosides, 5% light cream, 0.5% pectin, and the remainder being raw milk.

[0098] The preparation process of the above-mentioned dairy products is as follows:

[0099] S1: Pre-paste raw milk at 120℃ for 15 seconds;

[0100] S2: Heat the above raw milk to 50°C, then add the weighed steviol glycosides, light cream and pectin in sequence, mix and cut to obtain the mixture;

[0101] S3: The above mixture is first homogenized at a temperature of 70℃ and a pressure of 20MPa, and then sterilized at 95℃ for 300s. The outlet temperature during sterilization is 45℃ to obtain the fermentation base material.

[0102] S4: Pour the obtained fermentation base into a fermentation tank, then add the composition and mix evenly. Ferment at 45°C until the acidity reaches 65°T. After cooling to 15°C, aseptically bottle the fermented milk and store it in a 2°C cold storage.

[0103] Dairy products containing the compositions of Example 1 and Comparative Examples 1-7 were prepared according to the above method.

[0104] 2. Functional testing of dairy products

[0105] 2.1 Feed and Animal Sources

[0106] The basic feed (composed of the following percentages by weight: 20% wheat flour, 10% rice flour, 20% corn, 26% hulled wheat bran, 20% soybean, 2% fish meal, and 2% bone meal) was purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.

[0107] SPF-grade BALB / c mice (half male and half female, 6 weeks old, weighing 18-20g) were purchased from the Comparative Medicine Center of Yangzhou University. The experimental animals were housed in the experimental animal facility of the College of Food Science and Engineering, Yangzhou University. Laboratory temperature: 23.0±1.0℃, humidity: 50±5%, lighting conditions: cycled every 12 hours (8:00~20:00), with free access to water and food. All experimental conditions met the "Standards for Laboratory Animal Environment and Facilities" issued by the Ministry of Health of the People's Republic of China.

[0108] 2.2 Experimental Methods

[0109] SPF-grade BALB / c mice were acclimatized for one week and then divided into a control group, a model group, and an experimental group. Since there was no significant difference in body weight among the groups after grouping, the grouping was deemed appropriate. The mice in each group were treated as follows:

[0110] Control group and model group: 10 mice in each group were given 0.5 mL of sterile water by gavage twice a day.

[0111] Experimental group: Divided into groups 1-7, with 10 mice in each group, each mouse was given 0.5 mL of the dairy product prepared from the composition of Example 1 and Comparative Examples 1-7 by gavage twice a day.

[0112] After 14 days of continuous treatment, mice in each group were fasted but allowed free access to water for 16 hours. Then, mice in the model and experimental groups were administered compound diphenoxylate (5 mg / kg, bw) by gavage, while the control group was administered the same volume of distilled water by gavage. 30 minutes later, all mice in each group were administered gum arabic ink (containing 10% activated carbon powder and 10% activated gum arabic) by gavage. 30 minutes later, five mice from each group were randomly selected and euthanized by cervical dislocation. The portion of the intestine from the pylorus to the cecum was quickly removed, and the length of the intestinal segment was measured as the total small intestine length L0 (cm). The distance from the pylorus to the ink protrusion edge was measured as the ink propulsion length L1 (cm). The formula for calculating the small intestine ink propulsion rate is shown below:

[0113] Ink propulsion rate = L1 / L0 × 100%.

[0114] The remaining 5 mice in each group were administered ink via gavage, and the time of their first black feces, the number of fecal pellets excreted over 6 hours, and the total mass of feces were recorded. During the experiment, the mice had free access to food and water, and all mice in each group were fed standard diets. The enclosure was kept in a constant temperature and humidity environment, with a temperature of 25±2℃, a humidity of 50%±5%, and a light condition of 12 hours of light followed by 12 hours of darkness.

[0115] 2.4 Experimental results: as shown in Table 3.

[0116] Table 3. Effects of various dairy products on defecation in constipated mice.

[0117]

[0118]

[0119] As can be seen from the results in Table 3, compared with the dairy products of Comparative Examples 1-7, the dairy product of Example 1 of this invention has a more significant effect on improving the intestinal ink propulsion rate of mice, shortening the time to first black stool, and increasing the amount of stool in mice. These results indicate that the dairy product prepared from the composition of Example 1 has a good effect on promoting intestinal peristalsis and relieving constipation.

[0120] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the methods and techniques disclosed above without departing from the scope of the present invention to create equivalent embodiments. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A probiotic composition, characterized in that, Based on weight parts, its raw material composition includes: 1-5 parts of probiotic mixture, 10-20 parts of prebiotic mixture, and 1-3 parts of promoter; The probiotic mixture comprises Lactococcus lactis and Lactobacillus plantarum, wherein the mass ratio of Lactococcus lactis subsp. lactis to Lactobacillus plantarum is 1:(0.5~1); The prebiotic mixture comprises oligosaccharides, polyphenols, and protein hydrolysates, wherein the mass ratio of oligosaccharides, polyphenols, and protein hydrolysates is 1:(0.3~0.5):(0.5~1); The promoter comprises plant polysaccharides and chlorogenic acid, wherein the mass ratio of plant polysaccharides to chlorogenic acid is 1:(0.01~0.05).

2. The probiotic composition according to claim 1, characterized in that, The lactococcus lactis strain mentioned is Lactococcus lactis subsp. lactis WG-lla 26, with a viable count of 1×10⁻⁶. 8 ~1×10 10 CFU / g.

3. The probiotic composition according to claim 1, characterized in that, The *Lactobacillus plantarum* mentioned is *Lactobacillus plantarum* WG-lp40, with a viable count of 1×10⁻⁶. 8 ~1×10 10 CFU / g.

4. The probiotic composition according to claim 1, characterized in that, The oligosaccharide is selected from one or more of the following: fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, xylooligosaccharides, lactulose, and human milk oligosaccharides (HMOs).

5. The probiotic composition according to claim 1, characterized in that, The polyphenols are selected from one or more of curcumin, tea polyphenols, catechins, anthocyanins, and quercetin, or a mixture thereof.

6. The probiotic composition according to claim 1, characterized in that, The protein hydrolysates are quinoa protein peptides and / or walnut polypeptides.

7. The probiotic composition according to claim 1, characterized in that, The plant polysaccharides are selected from one or more of the following: red bean grass polysaccharide, black tea polysaccharide, ginseng polysaccharide, wolfberry polysaccharide, astragalus polysaccharide, ginseng polysaccharide, yam polysaccharide, rose polysaccharide, and inulin.

8. Use of a probiotic composition according to any one of claims 1-7 in the preparation of a medicine, food or health product for improving gut health.

9. A drug, food, or health product, characterized in that, The composition comprises the probiotic composition according to any one of claims 1-7.

10. The food product according to claim 9, characterized in that, The food is selected from at least one of dairy products, grain products, soy products, condiments, ice cream, and beverages.

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