Lactobacillus plantarum P16 for regulating intestinal flora and its application, product and method

Abstract

The present invention discloses a plant lactobacillus P16 for regulating intestinal flora and its application, product and method, which belong to the field of microbial technology. The present invention provides a plant lactobacillus P16 for regulating intestinal flora, and its deposit number is CCTCC NO: M 2023767. The present invention also provides the use of the plant lactobacillus P16 for preparing probiotics and / or preparing antibacterial agents and / or in vitro antibacterial agents, as well as probiotics, antibacterial agents and in vitro antibacterial methods. The plant lactobacillus P16 of the present invention can significantly inhibit the proliferation of pathogenic bacteria, can smoothly pass through the gastrointestinal tract of the body to exert probiotic effects, improve the health of the body, and the strain is sensitive to common antibiotics, has high safety, and has great potential in the application of products for regulating the balance of intestinal flora.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and specifically relates to a Lactobacillus plantarum P16 for regulating intestinal flora, and an application, a product and a method thereof. Background Art

[0002] The human intestine contains a large number of bacteria, including beneficial bacteria, neutral bacteria and harmful bacteria, among which beneficial bacteria are dominant. Their mutual relationship constitutes a dynamic balance, and maintaining the balance of the flora is the premise for maintaining the health of the body. Due to the influence of various adverse external factors, improper diet, old age and physical weakness, or acute and chronic diseases of the intestine and other systems, as well as long-term use of antibiotics and hormones, the balance of intestinal flora is destroyed. Changes in the flora force the reduction of beneficial bacteria and the increase of harmful bacteria, which trigger some physical diseases, impair the body's immune function and increase the probability of tumors; at the same time, the number of pathogenic bacteria in the intestine increases, causing infectious inflammation; the destruction of the balance of the flora will also reduce the synthesis of human nutrients such as vitamins and amino acids. For this kind of intestinal flora balance disorder, antibiotics are often used clinically to treat over-reproduction of pathogens, but at the same time, antibiotics will also inhibit the growth of beneficial bacteria, causing a new disorder system of intestinal flora; there are also ways to regulate the balance of intestinal flora by using traditional Chinese medicine. The research on the mechanism of Chinese medicine in treating intestinal flora balance diseases is still in its initial stage, and further research and exploration in terms of mechanism is needed.

[0003] A large number of studies have shown that oral probiotics can quickly change the structure of the host's intestinal flora. This is because a large number of live probiotics are directly delivered into the intestines, causing the structure of the intestinal flora to change in a short period of time. In addition, the short-chain fatty acids, hydrogen peroxide, and lactobacilli produced by the growth and metabolism of probiotics after entering the intestines can inhibit or kill pathogens, and can prevent the growth of harmful bacteria through competitive inhibition. Lactic acid bacteria are recognized as safe microorganisms for consumption. Digging out probiotics that are beneficial to human health, can inhibit the growth of harmful bacteria, have high stress resistance, can resist various digestive enzymes, can smoothly pass through the gastrointestinal tract, and have strong colonization ability can not only enrich the probiotic strain resources, but also have important significance for the development of human health products.

[0004] Lactobacillus plantarum is a kind of beneficial bacteria. Currently, the types of Lactobacillus plantarum that have been reported to regulate the balance of intestinal flora are quite limited and have single functions. There is an urgent need to develop more new strains of Lactobacillus plantarum that have both the function of regulating the balance of intestinal flora and a broad-spectrum antibacterial effect. Summary of the invention

[0005] In view of the insufficient number of available Lactobacillus plantarum species in the prior art and the need to develop more new strains of Lactobacillus plantarum, the present invention provides a Lactobacillus plantarum P16 for regulating intestinal flora and its application, product and method.

[0006] The technical solution of the present invention is as follows:

[0007] A Lactobacillus plantarum strain P16 for regulating the balance of intestinal flora, characterized in that its preservation number is CCTCC NO: M 2023767.

[0008] Use of Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M 2023767 in preparing probiotics and / or preparing antibacterial agents and / or in vitro bacteriostasis.

[0009] The probiotic agent is selected from: drugs for regulating intestinal flora or health foods for regulating intestinal flora;

[0010] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis;

[0011] Preferably, the in vitro antibacterial method comprises: co-culturing Lactobacillus plantarum P16 and intestinal pathogenic bacteria in vitro;

[0012] Preferably, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis;

[0013] Preferably, the intestinal flora regulation is selected from: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria;

[0014] Preferably, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index;

[0015] Preferably, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group;

[0016] Preferably, the beneficial intestinal bacteria include: Lachnospira, Verrucomicrobiales, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group, Candidatus_Saccharimonas;

[0017] Preferably, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

[0018] A probiotic comprises: an active ingredient; characterized in that the active ingredient comprises: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M 2023767.

[0019] The probiotic agent is selected from: drugs for regulating intestinal flora or health foods for regulating intestinal flora;

[0020] Preferably, the drug for regulating intestinal flora further comprises: pharmaceutically acceptable excipients;

[0021] Preferably, the health food for regulating intestinal flora further comprises: edible auxiliary materials.

[0022] The intestinal flora regulation is selected from: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria;

[0023] Preferably, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index;

[0024] Preferably, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group;

[0025] Preferably, the beneficial intestinal bacteria include: Lachnospira, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group, Candidatus_Saccharimonas;

[0026] Preferably, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

[0027] An antibacterial agent comprises an antibacterial active ingredient; the antibacterial active ingredient comprises: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M2023767.

[0028] The antibacterial agent further comprises: auxiliary materials;

[0029] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, and Streptococcus mitis.

[0030] The invention discloses an in vitro bacteriostasis method, which adopts Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M 2023767 to inhibit bacteria.

[0031] Co-culture Lactobacillus plantarum P16 with enteric pathogenic bacteria in vitro;

[0032] Preferably, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, and Streptococcus mitis.

[0033] The present invention provides a Lactobacillus plantarum strain P16, whose preservation number is: CCTCC NO: M 2023767; preservation time: May 17, 2023; preservation unit: China Center for Type Culture Collection; preservation address: China, Wuhan, Wuhan University.

[0034] The specific morphological characteristics of the plant lactobacillus P16 are as follows:

[0035] The bacterial characteristics are: Gram staining positive, rod-shaped cells with blunt ends, and orderly arrangement under an optical microscope.

[0036] The characteristics of the colonies are as follows: on solid MRS medium, the colonies are milky white or light yellow, convex, with a smooth and moist surface and neat edges.

[0037] The plant lactobacillus P16 provided by the invention has strong gastric juice and bile salt tolerance. After being treated with artificial gastric juice at pH 3.0 for 3 hours, its survival rate reaches 86.1%, and after being treated with 0.3% bile salt for 3 hours, its survival rate can reach 93.9%.

[0038] The invention screens out Lactobacillus plantarum P16 with the ability to inhibit Escherichia coli through an inhibition zone method, wherein the diameter of the inhibition zone for Escherichia coli reaches 25.4 mm, and further, the diameter of the inhibition zone for Salmonella enteritidis reaches 21.3 mm, the diameter of the inhibition zone for Staphylococcus aureus reaches 20.8 mm, the diameter of the inhibition zone for Clostridium perfringens reaches 23.5 mm, the diameter of the inhibition zone for Listeria monocytogenes reaches 15 mm, the diameter of the inhibition zone for Klebsiella pneumoniae reaches 18 mm, and the diameter of the inhibition zone for Streptococcus mitis reaches 28.5 mm. The invention is a probiotic with strong pathogen inhibition ability, can significantly reduce the growth of pathogens, and effectively alleviate the intestinal flora disorder caused by pathogens.

[0039] Furthermore, after co-culturing with Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes, Lactobacillus plantarum P16 had an inhibition rate of more than 99.9% on the three intestinal pathogens, indicating that Lactobacillus plantarum P16 can reduce the invasion of pathogens on the intestines and has the function of regulating the balance of intestinal microecology.

[0040] In an in vitro cell adhesion experiment, the Lactobacillus plantarum P16 can significantly inhibit the adhesion of Escherichia coli to human intestinal epithelial cells Caco-2, reduce the pathogenicity of pathogenic bacteria, and effectively improve intestinal health and enhance intestinal barrier function.

[0041] The Lactobacillus plantarum P16 is sensitive to 15 common antibiotics, indicating that Lactobacillus plantarum P16 is a safe probiotic that is not resistant to drugs.

[0042] At the same time, the plant lactobacillus P16 provided by the present invention showed a regulatory effect on the intestinal flora of mice in a mouse gavage experiment, improving the richness of the intestinal flora and the diversity of the community. The results of the analysis of the intestinal flora of mice showed that after the intervention of plant lactobacillus P16, the abundance of beneficial bacteria of Akkermansia (Akermansia) in the intestinal tract of mice was significantly higher than that of the DSS group, while the harmful bacteria Bacteroides (Bacteroides) and Helicobacter (Helicobacter) were significantly reduced.

[0043] The products include but are not limited to food or medicine. The food use form may include but is not limited to fermented dairy products, fermented bean products, fermented fruit and vegetable products, fermented meat products, fermented beverages, probiotic starter cultures, and probiotic solid beverages. The fermented dairy products include room temperature yogurt, low temperature yogurt, stirred yogurt, set yogurt, drinkable yogurt, cheese, lactic acid bacteria beverages, etc.

[0044] In some embodiments, the present invention provides a method for producing a composition for promoting the growth of bifidobacteria and / or lactic acid bacteria and / or regulating the balance of a subject's flora.

[0045] According to the embodiment, the product includes protein powder and capsules.

[0046] Beneficial effects of the present invention:

[0047] Compared with the prior art, the present invention provides a plant lactobacillus P16 with the ability to regulate the balance of intestinal flora and broad-spectrum antibacterial. The plant lactobacillus P16 of the present invention can smoothly pass through the gastrointestinal tract, form a biological barrier with a protective effect in the intestinal tract, reduce and repair the damage of invasive bacteria to the intestinal mucosa, and regulate the balance of intestinal flora. The strain has a strong pathogen inhibition ability, not only has a strong inhibitory effect on Escherichia coli, Salmonella enteritidis, and Staphylococcus aureus, but also has a strong inhibitory ability on the growth of Clostridium perfringens and Listeria monocytogenes, and can also effectively inhibit the growth of Streptococcus mitis and Klebsiella pneumoniae, and is a probiotic with an inhibitory effect on multiple pathogens; at the same time, when the strain is co-cultured with Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes, it can significantly inhibit the proliferation of pathogens, indicating that it helps to control the invasion of intestinal pathogens on the body, inhibit its growth, and regulate the balance of intestinal flora. This strain has a strong tolerance to gastric juice and bile salts, which means that the strain can smoothly pass through the gastrointestinal tract of the body to exert its probiotic effects and improve the health of the body. In addition, this strain is sensitive to common antibiotics and has high safety, and has great potential for application in products that regulate the balance of intestinal flora.

[0048] The deposit information of the Lactobacillus plantarum strain P16 of the present invention is as follows:

[0049] Deposit number: CCTCC NO:M 2023767;

[0050] Classification name: Lactobacillus plantarum P16

[0051] Latin name: Lactiplantibacillusplantarum P16;

[0052] Deposit date: May 17, 2023;

[0053] Depository: China Center for Type Culture Collection;

[0054] Deposit address: Wuhan University, Wuhan, China. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 This is the colony morphology of Lactobacillus plantarum P16 in Experimental Example 1 of the present invention.

[0056] Figure 2 The bacterial morphology of Lactobacillus plantarum P16 of Experimental Example 1 of the present invention observed under a microscope.

[0057] Figure 3 This is a bar graph showing the results of the gastric juice and bile salt tolerance experiment of Lactobacillus plantarum P16 in Experimental Example 2 of the present invention.

[0058] Figure 4 This is a photograph of the inhibition zone of Lactobacillus plantarum P16 against pathogenic bacteria in Experimental Example 3 of the present invention.

[0059] Figure 5 This is a line graph of the DAI score index of the mouse experiment in Experimental Example 7 of the present invention.

[0060] Figure 6 This is a bar graph of the live bacterial count detection experiment in mouse colon contents in Experimental Example 7 of the present invention.

[0061] Figure 7 This is a bar graph of the α-diversity analysis of the mouse intestinal flora in Experimental Example 7 of the present invention. The left graph is the ACE index, the middle graph is the Chao1 index, and the right graph is the Shannon index.

[0062] Figure 8 This is a bar graph of the relative abundance of the mouse intestinal flora level in Experimental Example 7 of the present invention.

[0063] Fig. 9 This is a bar graph of species difference analysis of different groups of mouse intestinal flora in Experimental Example 7 of the present invention.

[0064] Figure 5-Figure 9 The tags in are listed below:

[0065] NC represents the normal group mice gavaged with PBS;

[0066] DSS represents the model group mice that were given PBS intragastrically and then drank DSS freely;

[0067] P16 represents a group of mice treated with Lactobacillus plantarum, which were intragastrically administered with PBS containing the Lactobacillus plantarum P16 of the present invention and then freely drank DSS. DETAILED DESCRIPTION

[0068] The present invention is further described in detail below in conjunction with embodiments and experimental examples, but the embodiments of the present invention are not limited thereto, nor are they intended to limit the scope of protection of the present invention.

[0069] Sources of biological materials

[0070] 1. The deposit number of Lactiplantibacillus plantarum P16 used in Experimental Examples 1-7 of the present invention is CCTCC NO: M 2023767.

[0071] 2. The Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, and Listeria monocytogenes used in Experimental Example 3 or 4 of the present invention were all purchased from the Guangdong Provincial Microbiological Culture Collection Center. Streptococcus mitis is Streptococcus mitis strain 5013, which has been registered in NCBI with the accession number MT512114; Klebsiella pneumoniae is Klebsiella pneumoniae strain 587, which has been registered in NCBI with the accession number MT573143. Strain 5013 and strain 587 are currently preserved in the applicant's laboratory, and the applicant promises to release them to the public within 20 years from the date of application of the present invention for verification of the technical effects of the present invention.

[0072] 3. The human colorectal adenocarcinoma Caco-2 cells used in Experimental Example 5 of the present invention were purchased from China Center for Type Culture Collection.

[0073] 4. The SPF BALB / c male mice (19-20 g) used in Experimental Example 7 of the present invention were purchased from Wuhan Biotech Model Biotechnology Co., Ltd.

[0074] The first group of embodiments, Lactobacillus plantarum P16 of the present invention

[0075] This group of embodiments provides a plant lactobacillus (Lactiplantibacillus plantarum) strain P16 for regulating the balance of intestinal flora. All embodiments in this group have the following common features: the deposit number of the plant lactobacillus (Lactiplantibacillus plantarum) strain P16 is CCTCC NO: M 2023767.

[0076] Any act of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, or promising to sell a strain of Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M 2023767, and / or using a strain of Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M2023767 in combination with other probiotics, and / or using a strain of Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M 2023767 to antagonize Helicobacter pylori, preparing an antagonistic Helicobacter pylori product, and / or preparing an immunomodulatory product all fall within the scope of protection of the present invention.

[0077] The other probiotics include but are not limited to: Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii lactic acid subsp., Lactobacillus helveticus, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus salivarius, Saccharomyces cerevisiae, Delb's spore-forming torula, Candida, Wickham's yeast, Pichia, Saccharomyces boulardii, Pseudomonas, Saccharomyces cerevisiae ... The present invention also includes the following: Saccharomyces cerevisiae, Bacillus thuringiensis, Brevibacillus laterosporus, Bacillus megaterium, Bacillus colloidal, Bacillus azotobacterium, Bacillus sphaericus, Clostridium butyricum, Bifidobacterium adolescentis, Bifidobacterium angularis, Bifidobacterium animalis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium bovis, Bifidobacterium breve, Bifidobacterium dentate, Bifidobacterium infantis (i.e., Bifidobacterium longum infantis subsp. infantis), Bifidobacterium lactis (i.e., Bifidobacterium animalis subsp. lactis), Bifidobacterium longum, Bifidobacterium pseudocatenulatum, as well as Bifidobacterium thermophilum and Bifidobacterium acidothermophilum.

[0078] Those skilled in the art can make routine selections or adjustments to pharmaceutical excipients according to actual production needs, in combination with conventional technical means or basic common sense of pharmaceutical production processes (for example, "Encyclopedia of Preparation Technology", "Pharmaceutical Preparation Technology", etc.), and then prepare a Lactobacillus plantarum strain P16 with a preservation number of CCTCC NO: M 2023767 into products with different dosage forms, different storage conditions, and different shelf lives. This has no technical barriers for those skilled in the art and is possible and easy to do.

[0079] The second group of examples, use of Lactobacillus plantarum P16

[0080] The present group of embodiments discloses the use of Lactobacillus plantarum P16 with the deposit number of CCTCC NO: M 2023767 in the preparation of probiotics and / or antibacterial agents and / or in vitro bacteriostasis.

[0081] In some embodiments, the probiotic is selected from: drugs that regulate intestinal flora or health foods that regulate intestinal flora;

[0082] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis;

[0083] Preferably, the in vitro antibacterial method comprises: co-culturing Lactobacillus plantarum P16 and intestinal pathogenic bacteria in vitro;

[0084] Preferably, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis;

[0085] Preferably, the intestinal flora regulation is selected from: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria;

[0086] Preferably, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index;

[0087] Preferably, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group;

[0088] Preferably, the beneficial intestinal bacteria include: Lachnospira, Verrucomicrobiales, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group, Candidatus_Saccharimonas;

[0089] Preferably, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

[0090] The third group of embodiments, the probiotic agent of the present invention

[0091] This group of embodiments provides a probiotic agent. All embodiments of this group have the following common features: the probiotic agent comprises: an active ingredient; the active ingredient comprises: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M2023767.

[0092] In some embodiments, the probiotic is selected from: drugs that regulate intestinal flora or health foods that regulate intestinal flora;

[0093] Preferably, the drug for regulating intestinal flora further comprises: pharmaceutically acceptable excipients;

[0094] In a more specific embodiment, the pharmaceutically acceptable excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0095] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the Lactobacillus plantarum strain P16 with a deposit number of CCTCC NO: M 2023767 into different dosage forms, such as powders, tablets, injections, oral liquids, suppositories, gels, patches, sprays, lotions, granules, etc.

[0096] In a specific embodiment, the dosage form of the drug is selected from: one or more of powder, tablet, liquid, and capsule.

[0097] Preferably, the health food for regulating intestinal flora further comprises: edible auxiliary materials.

[0098] In a more specific embodiment, the edible excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0099] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Food Technology", "Food Processing Technology", "Health Food Research and Development and Production", "Health Food GMP Implementation and Management", "Functional Food Tutorial", "Functional Food Science", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the Lactobacillus plantarum strain P16 with a preservation number of CCTCC NO: M 2023767 into different dosage forms, such as capsules, tablets, lozenges, oral liquids, powders, granules, etc.

[0100] In a specific embodiment, the dosage form of the product is selected from: one or more of capsules, tablets, lozenges, oral liquids, powders, and granules.

[0101] In other embodiments, the intestinal flora is regulated by: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria;

[0102] Preferably, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index;

[0103] Preferably, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group;

[0104] Preferably, the beneficial intestinal bacteria include: Lachnospira, Verrucomicrobiales, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group, Candidatus_Saccharimonas;

[0105] Preferably, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

[0106] The fourth group of examples, the antibacterial agent of the present invention

[0107] This group of embodiments provides an antibacterial agent. All embodiments in this group have the following common features: the antibacterial agent includes an antibacterial active ingredient; the antibacterial active ingredient includes: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M2023767.

[0108] In a further embodiment, the antimicrobial agent further comprises: an auxiliary material;

[0109] In a more specific embodiment, the excipient is selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0110] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of drug preparation (for example, "Encyclopedia of Preparation Technology", "Drug Preparation Technology", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare the Lactobacillus plantarum strain P16 with a deposit number of CCTCC NO: M 2023767 into different dosage forms, such as powders, tablets, injections, oral liquids, suppositories, gels, patches, sprays, lotions, granules, etc.

[0111] In a specific embodiment, the dosage form of the antibacterial agent is selected from: one or more of powder, tablet, liquid, and capsule.

[0112] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, and Streptococcus mitis.

[0113] The fifth group of embodiments, the in vitro antibacterial method of the present invention

[0114] This group of embodiments provides an in vitro antibacterial method. All embodiments in this group have the following common features: Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M 2023767 is used for antibacterial.

[0115] In a specific embodiment, Lactobacillus plantarum P16 and intestinal pathogenic bacteria are co-cultured in vitro;

[0116] Preferably, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, and Streptococcus mitis.

[0117] The following experimental examples 1-7 used the Lactobacillus plantarum culture medium:

[0118] LBS medium formula: 5.0g yeast extract powder, 10.0g tryptone, 6.0g potassium dihydrogen phosphate, 0.034g ferrous sulfate, 0.575g magnesium sulfate, 20.0g glucose, 25.0g sodium acetate, 2.0g ammonium citrate, 0.12g manganese sulfate, after adjusting the pH value to 5.5, add 1ml Tween-80 and 1.3ml glacial acetic acid, heat and stir to dissolve in 1000ml distilled water, and sterilize at 118℃ for 15min. Add 15.0g agar to the solid medium.

[0119] The preparation method of MRS culture medium refers to GB4789.35-2016, and 1.5% agar is added to the solid culture medium.

[0120] Modified MRS medium: peptone 10 g, beef extract 10 g, yeast extract 5 g, glucose 20 g, K2HPO4 2 g, sodium citrate 2 g, MgSO4·7H2O 0.2 g, MnSO4·4H2O 0.2 g, Tween 80 1 ml, and distilled water 1000 ml.

[0121] L-MRS medium: peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, K2HPO42g, sodium citrate 2g, sodium acetate 5g, MgSO4·7H2O 0.2g, MnSO4·4H2O 0.2g, Tween 80 1ml, L-cysteine ​​hydrochloride 1g, 1000ml distilled water.

[0122] Clostridium perfringens culture medium: BHI culture medium was used for activation culture medium, purchased from Qingdao Haibo; solid culture medium TSA was used for inhibition zone experiment.

[0123] Listeria monocytogenes culture medium: solid culture medium using BHI plus 5% defibrinated sheep blood, purchased from Qingdao Haibo.

[0124] Escherichia coli, Salmonella enteritidis, Staphylococcus aureus culture medium: LB medium

[0125] Unless otherwise specified, all kinds of reagents and consumables used in the embodiments and experimental examples of the present invention are commercially available, and the relevant experimental steps are common operations in the field and have technical meanings that can be conventionally understood by those skilled in the art.

[0126] Experimental Example 1: Isolation, screening and identification of Lactobacillus plantarum P16

[0127] (1) Isolation and screening of strains

[0128] Collect kimchi samples, place them in sterile sampling tubes and transport them in ice boxes, dilute them with 0.85% saline gradients under sterile conditions, select appropriate dilution gradients and spread them on LBS agar plates, and culture them at 37°C for 48-72h. Observe the colony morphology with the naked eye to pick out suspected single colonies, observe them under a microscope, and perform preliminary screening and purification culture. After purification, use MRS liquid to culture anaerobically at 37°C for 8-12h, centrifuge to remove the supernatant, and resuspend in a sterilized 30% glycerol aqueous solution.

[0129] (2) Morphological characteristics and 16S rDNA identification

[0130] The colony and bacterial morphology of Lactobacillus plantarum P16 are shown in Figure 1 , Figure 2 The single colony of Lactobacillus plantarum P16 on the MRS solid plate is round, medium-sized, with neat edges, milky white or light yellow. The bacterial body morphology is Gram-positive rod, short rod, single rod or arranged, with blunt ends.

[0131] The screened target strains were cultured in liquid, the cells were collected, the genomic DNA was extracted, and the universal primers 27F and 1492R recorded in paragraph 58 of Chinese invention patent ZL202210478937.4 were used for PCR amplification reaction. The PCR amplification procedure is: 94°C pre-denaturation for 5 minutes, 94°C denaturation for 30 seconds, 55°C annealing for 30 seconds, 72°C extension for 90 seconds, a total of 35 cycles; 72°C extension for 10 minutes. The content and purity of the PCR amplification product were then tested, and after passing the test, it was sent to Wuhan Jinkairui Bioengineering Co., Ltd. for sequencing. According to the sequencing results, the BLAST tool in the NCBI database was used for homology comparison, and the obtained strain was identified as Lactobacillus plantarum. The strain was named P16 and sent for preservation. Its preservation information is as follows:

[0132] Deposit number: CCTCC NO:M 2023767;

[0133] Classification and nomenclature: Lactobacillus plantarum P16;

[0134] Latin name: Lactiplantibacillusplantarum P16;

[0135] Depository: China Center for Type Culture Collection;

[0136] Address of depository: Wuhan University, Wuhan, China

[0137] Date of preservation: May 17, 2023.

[0138] Experimental Example 2: Tolerance of Lactobacillus plantarum P16 to Simulated Digestive Fluid

[0139] Artificial gastric juice: prepare PBS solution, add 0.3% pepsin, adjust the pH to 3.0 with 1 mol / l HCl, and filter with 0.22 μm microporous filter membrane for sterilization after fully dissolving and set aside.

[0140] Artificial bile salt solution: add 0.2% (w / v) sodium thioglycolate to MRS liquid culture medium, then add 0.3% ox bile salt, fully dissolve and filter with a 0.22 μm microporous filter membrane for sterilization.

[0141] The preserved strain was activated twice, 1 ml of the bacterial solution cultured for 12 h was centrifuged (10,000 × g, 5 min, 4 °C), washed twice with PBS, and the bacteria were added to 10 ml of simulated artificial gastric juice and bile salt solution, respectively, and then incubated at 37 °C for 3 h. Samples were taken at 0 h and 3 h, diluted and spread in MRS solid culture medium, and the process was repeated three times. The cells were cultured in an incubator at 37 °C for 48 h and then counted.

[0142] The tolerance results of Lactobacillus plantarum P16 in artificial gastric juice and artificial bile salt solution are shown in Figure 3 .Depend on Figure 3 It can be seen that the strain maintained a very high survival rate after being treated in artificial gastric juice and bile salt for 3 hours, indicating that the strain has good gastrointestinal fluid tolerance.

[0143] Experimental Example 3: Antagonistic effect of Lactobacillus plantarum P16 on pathogens

[0144] Preparation of Lactobacillus plantarum P16 bacterial solution: inoculate Lactobacillus plantarum P16 into liquid MRS culture medium at a certain ratio, and culture at 37° C. overnight to obtain antagonistic bacterial solution.

[0145] Preparation of indicator bacteria suspension: Escherichia coli, Salmonella enteritidis, and Staphylococcus aureus were activated on LB solid plate medium, cultured at 37°C for 24 h, and the bacterial moss was picked up and placed in physiological saline to prepare bacterial suspension for 10 8 CFU / ml; Clostridium perfringens was activated in BHI solid medium, cultured at 37°C for 24 h, and the concentration of bacterial suspension was adjusted to 10 8 CFU / ml; Listeria monocytogenes was activated in a solid plate medium containing BHI plus 5% sheep blood, cultured at 37°C for 24 h, and the cells were collected to prepare a suspension. The concentration of the bacterial suspension was adjusted to 10 8 CFU / ml; Klebsiella pneumoniae was cultured in BHI solid medium with 5% sheep blood at 37°C for 16 h, and the concentration of bacterial suspension was adjusted to 10 8 CFU / ml; Streptococcus mitis was cultured in BHI solid medium with 5% sheep blood at 37°C for 16 h, and the concentration of bacterial suspension was adjusted to 10 8 CFU / ml.

[0146] Bacteriostatic experiment: Cool the culture medium corresponding to each pathogen to about 55°C, mix it with the indicator strain suspension in a certain proportion, and adjust the indicator bacteria concentration to 10 6 CFU / ml, and then quickly pour it into the plate with the Oxford cup placed in advance. After the culture medium cools and solidifies, take out the Oxford cup and add 200 μl of antagonistic bacteria solution (10 8 CFU / ml), 200 μl sterile MRS liquid culture medium was used as blank control, after standing for a few minutes, it was placed in a 37°C constant temperature incubator, and observed after 24 hours of incubation. The diameter of the inhibition zone was measured with a vernier caliper. Each bacterium was repeated three times. The results are shown in Table 1 and Figure 4 .

[0147] Table 1. Antagonistic ability of P16 against pathogens

[0148]

[0149]

[0150] The results are shown in Table 1 and Figure 4As shown, Lactobacillus plantarum P16 has a strong inhibitory ability against Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, and Clostridium perfringens, and also shows a strong inhibitory ability against Listeria monocytogenes and Klebsiella pneumoniae. Among them, the inhibitory ability of Lactobacillus plantarum against Streptococcus mitis reached 28.5mm.

[0151] Experimental Example 4: Antibacterial experiment of co-culture of Lactobacillus plantarum P16 and intestinal pathogenic bacteria

[0152] Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes were activated in the corresponding culture medium, and the bacterial count was adjusted to 1×10 8 CFU / ml. Escherichia coli and Salmonella enteritidis were inoculated into modified MRS liquid culture medium at a 1% inoculation rate, and Listeria monocytogenes was inoculated into BHI liquid culture medium at a 1% inoculation rate. The same amount of Lactobacillus plantarum P16 was inoculated into the culture medium with pathogens and mixed and cultured. At the same time, Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes were inoculated into the corresponding liquid culture medium with the same inoculation rate as control cultures. After static culture at 37°C for 24 hours, the inhibition of Lactobacillus plantarum P16 on pathogens was observed after co-culture with pathogens. The results are shown in Table 2.

[0153] Table 2. Co-culture experiment of P16 and intestinal pathogenic bacteria

[0154]

[0155] The results are shown in Table 2. After co-culture with pathogenic bacteria, the inhibition rate of Lactobacillus plantarum P16 on three intestinal pathogenic bacteria reached more than 99.9%, indicating that Lactobacillus plantarum P16 can reduce the invasion of pathogenic bacteria on the intestine and has the function of regulating the balance of intestinal microecology.

[0156] Experimental Example 5: Lactobacillus plantarum cell adhesion experiment

[0157] Cell culture: The revived Caco-2 cells were inoculated into 12-well cell culture plates in DMEM medium supplemented with 10% (v / v) fetal bovine serum. The number of cells added was 2.4 × 10 cells per well. 5 The cells were cultured at a constant temperature of 5% CO2 and 37°C until 80% of the cells adhered to the wall, and then washed three times with PBS for later use.

[0158] Preparation of Lactobacillus plantarum P16 bacterial solution: inoculate the secondary activated Lactobacillus plantarum P16 into liquid MRS medium at a 2% inoculum amount, culture at 37°C overnight to obtain bacterial solution, take 1 ml of the cultured bacterial solution, centrifuge at 10000 r / min at room temperature for 1.5 min to collect the bacteria, and wash twice with an equal amount of sterile PBS, add 1 ml of DMED medium to each of the precipitates after centrifugation, take the mixed bacterial solution and adjust the bacterial concentration to 10 8 CFU / ml, obtain the spare bacterial suspension and detect the number of viable bacteria.

[0159] Adhesion ability determination: remove the culture medium from the prepared Caco-2 monolayer, add PBS buffer to rinse twice, remove the buffer, add 1 ml / well of the prepared bacterial suspension, and incubate for 2 hours at 5% CO2 and 37°C. After the incubation, carefully remove the culture supernatant, add sterile PBS to rinse three times to remove non-adherent bacteria; add 0.2 ml / well of trypsin cell digestion solution, digest in a 37°C incubator for 5 minutes, take out and neutralize with 0.8 ml PBS, blow and rinse several times to make the system uniform, take out 10 μl of the mixed solution and count the cells with a hemocytometer (count number × 10 4 / ml), and the remaining suspension was used for viable bacteria counting.

[0160] The calculation formula of cell adhesion rate is:

[0161] Adhesion rate (%) = number of bacteria adhering to cells (CFU) / total number of bacteria added to the well x 100%

[0162] Adhesion capacity (CFU / cells) = number of bacteria adhering to cells (CFU) / number of cells in the well

[0163] Table 3. Adhesion test of Lactobacillus plantarum P16 to Caco-2 cells

[0164]

[0165] The adhesion of probiotics to intestinal epithelial cells is the basis for their probiotic function. The adhesion rate of Lactobacillus plantarum P16 to Caco-2 cells reached 13.1%, indicating that the strain can effectively colonize in the intestine after passing through the gastrointestinal tract and play a role in regulating the balance of intestinal flora.

[0166] Experimental Example 6: Antibiotic sensitivity test of Lactobacillus plantarum P16

[0167] The antibiotic sensitivity test was performed using the KB (susceptibility paper agar diffusion method). After the bacteria to be tested were activated, a bacterial suspension was prepared and the concentration of the bacterial suspension was adjusted to 10 8CFU / ml, take 200μl of bacterial suspension on the surface of MRS solid plate, evenly spread the bacterial liquid on the plate with a sterile cotton swab, stick the antibiotic sensitive paper on the MRS solid medium with sterile tweezers, mark it, and culture it at 37℃ for 24h, and measure the diameter of the inhibition zone with a vernier caliper. The edge of the inhibition zone is bounded by the fact that the plant lactobacillus can not be seen to grow obviously. The sensitivity results of the plant lactobacillus P16 strain to 12 commonly used antibiotics are shown in Table 4. The plant lactobacillus P16 is sensitive to these drugs, indicating that the strain is safe and can be potentially used in biological products or dietary supplements.

[0168] Table 4. Susceptibility of Lactobacillus plantarum P16 to antibiotics

[0169]

[0170] Note: S-susceptible; I-intermediately resistant; R-resistant

[0171] Experimental Example 7: The ability of Lactobacillus plantarum P16 to regulate the balance of intestinal flora in mice

[0172] The animal model was induced by DSS. Thirty SPF BALB / c male mice (19-20g) were selected for the experiment. The room temperature of the animals was 23±2℃, the humidity was 50%±10%, the artificial light was 12h / d, and the diet was free. All mice were divided into 3 groups, 10 in each group, namely the normal group (NC), the model group (DSS), and the Lactobacillus plantarum treatment group (P16). The specific experimental process is as follows: one week of adaptive feeding, in the second week, all mice except the normal group were free to drink 3.5% DSS solution for 7 days, and the DSS solution was replaced every 2 days to ensure the concentration and quality of the DSS solution. At the same time, the experimental mice were gavaged with 0.2ml of the corresponding solution at regular intervals every day. The normal group (NC) was gavaged with PBS, the model group (DSS) was gavaged with PBS, and the Lactobacillus plantarum treatment group (P16) was gavaged with 5×10 9 CFU / ml of Lactobacillus plantarum P16 in PBS. The model group (DSS) and the Lactobacillus plantarum treatment group (P16) freely drank 3.5% DSS solution for 7 days, and then replaced with normal drinking water. Gavage was performed once a day at a fixed time for 14 consecutive days, with a gavage volume of 0.2ml each time; the normal group and the model group were gavaged with an equal volume of PBS every day. After the experiment, the mice were killed by cervical dislocation, and the abdomen was disinfected and dissected to obtain the contents of the cecum and colon of each group of mice.

[0173] 1) Disease Activity Index Evaluation. During the experiment, the health status, hair, body weight, food intake and feces of the mice were recorded every day, and the disease activity index (DAI) was evaluated. DAI score = (weight loss rate score + feces score + occult blood score) / 3. The DAI score is calculated as follows:

[0174] Table 5. DAI scoring criteria

[0175]

[0176] Depend on Figure 5 It can be seen that the DAI score of the DSS model group continued to increase from day 2 to day 5, and on day 5, the DAI score almost reached 4. The DAI score of the P16 group was significantly lower than that of the DSS model group, and its DAI score was less than 2, indicating that Lactobacillus plantarum P16 can alleviate the increase in the DAI score of mice and is beneficial to maintaining the health of mice.

[0177] 2) Experiment on detection of viable bacteria count in mouse colon contents. Weigh 1g of colon contents and mix them evenly in 9ml of normal saline, dilute them tenfold, and perform viable bacteria count detection on MRS culture medium. The colony count method is based on GB4789.2-2010 National Food Safety Standard for Food Microbiology Test for Determination of Total Colony Count.

[0178] Depend on Figure 6 It can be seen that the number of live bacteria in the colon contents of the model group mice was significantly lower than that of the NC group, while the number of live bacteria in the colon contents of the P16 group gavaged with Lactobacillus plantarum increased significantly. This shows that DSS reduces the number of intestinal microorganisms, while Lactobacillus plantarum P16 has the effect of enriching the number of microorganisms in the intestine and has a beneficial effect on the balance of the flora.

[0179] 3) 16S rDNA sequencing and analysis of mouse cecal contents. The results showed that Figure 7 As shown in the figure, compared with the normal group, the ACE index, Chao1 index and Shannon index of intestinal microbial α diversity in the DSS group were all decreased, indicating that DSS would cause a decrease in the α diversity index of the intestinal flora in mice; compared with the DSS group, the ACE index, Chao1 index and Shannon index of intestinal microbial α diversity in the P16 group were all increased, indicating that the intervention of Lactobacillus plantarum P16 would promote the increase of the α diversity index of the intestinal flora in mice, proving that supplementation with Lactobacillus plantarum P16 can improve the richness and community diversity of intestinal microbial flora.

[0180] Figure 8 The structure and differences of the phylum level of the intestinal flora of mice are shown. Compared with the DSS group, after intervention with Lactobacillus plantarum P16, the Firmicutes in the P16 group increased, the Bacteroidetes decreased, and especially the Verrucomicrobia significantly increased. This indicates that after intervention with Lactobacillus plantarum P16, the ratio of Firmicutes to Bacteroidetes can be increased, and this ratio may be related to the relief of intestinal inflammation in mice.

[0181] Fig. 9The genera showing changes in the dominant flora in the intestines of mice after intervention with Lactobacillus plantarum P16 showed that the abundance of unclassified-Lachnospiraceae, Akkermansia, Ruminococcaceae, Lachnospiraceae_NK4A136_group, and Candidatus_Saccharimonas was significantly higher than that in the DSS group. Among them, unclassified-Lachnospiraceae exists in the intestines of most healthy people and may be a potential beneficial bacterium that participates in the metabolism of a variety of carbohydrates, and the acetic acid and butyric acid produced by fermentation provide energy for the host. Studies have shown that Akkermansia plays a potential protective role in the development of inflammatory bowel disease. The reduced abundance of Akkermansia in the intestine may be a potential marker of IBD. Supplementation of Akkermansia can regulate intestinal microecological imbalance, improve the barrier function of the intestinal mucosa, and reduce inflammatory responses in the blood and intestines. Ruminococcaceae produces short-chain fatty acids, formic acid and acetic acid. Lachnospiraceae_NK4A136_group is considered a probiotic that produces butyrate during proliferation, and its abundance level is reported to be negatively correlated with intestinal inflammation. Candidatus_Saccharimonas may be an important factor in immune recovery in immunodeficient patients. Compared with the DSS group, the abundance of the harmful microbial genus Colidextribacter (Escherichia coli) decreased in the P16 group, and Bacteroides (Bacteroides) and Helicobacter (Helicobacter) decreased significantly.

[0182] The above intestinal flora analysis results show that DSS will reduce the diversity of intestinal microorganisms, enrich harmful bacteria, and thus lead to intestinal flora disorders; while intervention with Lactobacillus plantarum P16 can restore the diversity of intestinal microorganisms, inhibit the growth of pathogens, and promote the enrichment of beneficial bacteria, thereby regulating the balance of intestinal flora and restoring the flora homeostasis system.

Claims

1. A plant lactobacillus that regulates the balance of intestinal flora ( Lactiplantibacillus plantarum ) strain P16, characterized in that Its deposit number is CCTCC NO: M 2023767.

2. Use of Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M 2023767 in the preparation of probiotics and / or antibacterial agents and / or in vitro bacteriostasis.

3. The use of Lactobacillus plantarum P16 with a deposit number of CCTCC NO: M 2023767 according to claim 2 in the preparation of probiotics and / or antibacterial agents and / or in vitro bacteriostasis, characterized in that: The probiotic agent is selected from: drugs for regulating intestinal flora or health foods for regulating intestinal flora; and / or, the antimicrobial spectrum of the antimicrobial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis; And / or, the in vitro antibacterial activity includes: co-culturing Lactobacillus plantarum P16 and intestinal pathogenic bacteria in vitro; And / or, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis; And / or, the intestinal flora regulation is selected from: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria; And / or, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index; And / or, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group; And / or, the beneficial intestinal bacteria include: Lachnospira, Verrucomicrobiales, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group , Candidatus_Saccharimonas ; And / or, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

4. A probiotic comprising: Active ingredient; characterized in that the active ingredient includes: Lactobacillus plantarum P16 with a deposit number of CCTCCNO: M 2023767.

5. A probiotic according to claim 4, characterized in that: The probiotic agent is selected from: drugs for regulating intestinal flora or health foods for regulating intestinal flora; And / or, the drug for regulating intestinal flora further comprises: pharmaceutically acceptable excipients; And / or, the health food for regulating intestinal flora further comprises: edible auxiliary materials.

6. A probiotic according to claim 5, characterized in that: The intestinal flora regulation is selected from: increasing the intestinal microbial diversity index, alleviating the increase of DAI score, increasing the abundance of beneficial intestinal bacteria, and reducing the abundance of harmful intestinal bacteria; And / or, the intestinal microbial diversity index includes: ACE index, Chao1 index, Shannon index; And / or, the alleviating the increase in DAI score refers to: alleviating the increase in DAI score of mice in the DSS model group; And / or, the beneficial intestinal bacteria include: Lachnospira, Verrucomicrobiales, Akkermansia, Ruminococcus, Lachnospiraceae_NK4A136_group , Candidatus_Saccharimonas ; And / or, the harmful intestinal bacteria include: Escherichia coli, Bacteroides, and Helicobacter.

7. An antibacterial agent comprising an antibacterial active ingredient; characterized in that: The antibacterial active ingredients include: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M 2023767.

8. The antibacterial agent according to claim 7, characterized in that Also includes: Excipients; And / or, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, and Streptococcus mitis.

9. An in vitro antibacterial method, characterized in that: Lactobacillus plantarum P16 with a preservation number of CCTCC NO: M 2023767 was used for bacteriostasis.

10. The in vitro antibacterial method according to claim 9, characterized in that: Lactobacillus plantarum P16 was co-cultured with intestinal pathogenic bacteria in vitro; And / or, the intestinal pathogenic bacteria are selected from: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Clostridium perfringens, Listeria monocytogenes, Klebsiella pneumoniae, Streptococcus mitis.

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