Antibacterial composition against pathogenic bacteria comprising culture broth of streptococcus salivarius g7 and lactobacillus plantarum gspe05

Abstract

The present invention relates to an antibacterial composition against pathogenic bacteria, the composition comprising streptococcus salivarius g7 and lactobacillus plantarum gspe05, and specifically, exhibits high antibacterial activity against one or more bacteria selected from the group consisting of Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, Streptococcus mutans, Porphyromonas gingivalis, and Moraxella catarrhalis, and thus can contribute to the improvement of intestinal diseases, oral diseases, and respiratory diseases caused by these pathogenic strains.

Description

Antimicrobial composition against pathogenic bacteria comprising cultures of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05

[0001] The present invention relates to an antimicrobial composition against pathogenic bacteria comprising a culture solution of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05.

[0002] Many microorganisms grow on the mucous membranes of our body that come into contact with the outside world, and some exist in symbiotic relationships that exert mutually beneficial effects. The surface of the intestinal mucosa is the widest connection site between our body and the intestinal microorganisms. The intestinal microbiome consists of over 1,000 species of microorganisms and forms a unique microbial ecosystem; it is known to be metabolically interconnected and subject to natural selection at the host level. This intestinal microbiome performs functions such as stimulating the intestinal immune system, regulating intestinal epithelial cell proliferation, synthesizing essential amino acids, and increasing resistance to pathogens.

[0003] In addition, symbiotic microorganisms present in the nasal mucosa are known to inhibit pathogenic bacteria and viruses that cause respiratory infections. When Staphylococcus epidermidis cultured from the nasal mucosa of healthy individuals was transplanted into the nasal mucosa of mice and subsequently infected with the influenza virus, the viral load decreased by more than 90%, resulting in increased resistance to influenza infection, whereas mice without the transplanted Staphylococcus epidermidis developed fatal lung infections. This is because when symbiotic microorganisms inhabit the nasal mucosa stably, they exhibit a protective effect against external pathogens.

[0004] In addition to this, microbial communities formed in body parts that come into contact with the outside, such as the oral microbiome and the vaginal microbiome, increase resistance to pathogens and have a direct impact on the host's health. If the microbial composition of these communities changes or beneficial bacteria decrease, homeostasis is disrupted and the host becomes vulnerable to pathogen infection.

[0005] Accordingly, probiotics utilizing functional lactic acid bacteria have been developed to form a beneficial microbial community by transplanting beneficial bacteria into body parts that come into contact with the outside environment. These probiotics are known to have effects such as inhibiting the proliferation of harmful bacteria in the intestines to regulate immunity to normal levels, improving autoimmune skin diseases like atopic dermatitis and psoriasis, suppressing hypertension and metabolic syndrome, and lowering insulin resistance. Additionally, probiotics are known to exhibit antimicrobial effects against pathogens by secreting antimicrobial substances such as bacteriocins (antimicrobial peptides), short-chain fatty acids, and hydrogen peroxide. However, the strain specificity of the antimicrobial action of these beneficial bacteria against pathogens and the mechanism of their antimicrobial activity are not yet precisely known, and in particular, the antimicrobial action against specific pathogens through a specific combination of beneficial bacterial species has not yet been identified.

[0006] The object of the present invention is to provide an antimicrobial composition against pathogenic bacteria comprising cultures of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05.

[0007] To solve the above problem, the present invention provides an antimicrobial composition comprising a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution as an active ingredient.

[0008] The above mixture of cultures of Streptococcus salivarius G7 (accession number KCCM13161P) and Lactobacillus plantarum GSPE05 (KCTC 15128BP) may be a mixture obtained by culturing Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, under aerobic conditions at 37°C for 18 hours using Tryptic soy broth (TSB) + 0.5% glucose or MRS broth, and mixing the cultures of each in a volume ratio of 1:0.1 to 10. Preferably, the mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution may be a mixture obtained by mixing culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, cultured under aerobic conditions at 37°C for 18 hours using TSB+0.5% glucose or MRS broth, in a volume ratio of 1:1 to 7. If the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 deviates from the 1:1 to 7 ratio, the antimicrobial activity is significantly reduced. Most preferably, it is a mixture mixed in a volume ratio of 1:5.

[0009] The culture conditions of the above-mentioned Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 may be cultured at the same temperature and for the same period, or cultured under aerobic conditions at 37°C for 18 hours using TSB+0.5% glucose or MRS broth. The above-mentioned same culture conditions are intended to ensure a constant production of antimicrobial substances so that the antimicrobial substances produced by Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 during culture are combined in a constant amount. The type of culture medium is not limited to any specific type as long as it provides sufficient nutrients for the growth of the strains, and the culture temperature and period are also not limited.

[0010] In addition, the culture solution of the strain can be dried to produce a powder, and the produced powder can be mixed to prepare the culture solution mixture of the present invention. The mixing ratio may be a mixture in which the dried powders of the culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 are mixed in a weight ratio of 1:1 to 7. Most preferably, it is a mixture in which the dried powders of the culture solutions are mixed in a weight ratio of 1:5.

[0011] The present invention provides an antimicrobial composition against pathogenic bacteria comprising 11.5% by weight of Streptococcus salivarius G7 and 57.5% by weight of dried culture powder of Lactobacillus plantarum GSPE05.

[0012] The above antimicrobial activity refers to antimicrobial activity against one or more bacteria selected from the group consisting of Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, Streptococcus mutans, Porphyromonas gingivalis, and Moraxella catarrhalis.

[0013] In this invention, 'Bacillus cereus' is a Gram-positive, aerobic, and large bacillus. Although it usually does not cause symptoms of food poisoning or disease when present in food at a concentration of 10 CFU / g or less, an environment is created where this bacterium can grow well, resulting in 10 6 It is a pathogenic bacterium that causes diarrhea and vomiting when the CFU / g level exceeds 100%.

[0014] In this invention, 'Listeria monocytogenes' is a pathogenic bacterial species that causes listeriosis infection, and it infects pregnant women (fetuses), newborns, infants, the elderly, and people with underlying diseases who are vulnerable to infection, causing meningitis, sepsis, or miscarriage.

[0015] In this invention, 'Salmonella typhimurium' is a pathogenic Gram-negative bacterium found in the intestinal lumen that causes gastroenteritis and diarrhea in humans and other mammals, and exhibits symptoms similar to typhoid fever.

[0016] In the present invention, 'Streptococcus mutans' is also known as a caries bacterium and is one of the main causative bacteria of tooth decay. S. mutans causes tooth decay, discoloring the teeth black and causing them to rot.

[0017] In the present invention, 'Porphyromonas gingivalis' is a non-motile, Gram-negative, rod-shaped, anaerobic, pathogenic bacterium and is a representative bacterium that causes periodontitis. P. gingivalis is found in the oral cavity, upper gastrointestinal tract, respiratory tract, and colon in cases of periodontitis, and is known to be associated with Alzheimer's disease and rheumatoid arthritis.

[0018] In the present invention, 'Moraxella catarrhalis' is a non-motile, Gram-negative aerobic bacterium that can cause infections in the human respiratory system, ears, eyes, central nervous system, joints, etc., and is a major cause of bronchopulmonary infections. In particular, it causes bacterial pneumonia in adults with weakened immune systems and causes acute sinusitis, maxillary sinusitis, bacteremia, meningitis, conjunctivitis, chronic bronchitis, acute laryngitis in adults, and acute otitis media in children.

[0019] The present invention provides a pharmaceutical composition for treating or preventing intestinal diseases comprising, as an active ingredient, a mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

[0020] The above intestinal disease may be any one of the following: irritable bowel syndrome, ulcerative colitis, intestinal lesions associated with Crohn's disease or Behcet's disease, hemorrhagic rectal ulcer, and ileal pouchitis.

[0021] The present invention provides a composition for improving intestinal health comprising a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution as an active ingredient.

[0022] The above composition for improving intestinal health may be one or more formulations selected from the group consisting of tablets, capsules, pills, granules, liquid, powder, flakes, paste, syrup, gel, jelly, gums, and candies.

[0023] The present invention provides a pharmaceutical composition for the treatment or prevention of oral diseases comprising, as an active ingredient, a mixture of a culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and a culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

[0024] The above oral disease may be one or more selected from the group consisting of gingivitis, periodontitis, dental caries, and periapical disease caused by the above oral harmful bacteria, but is not limited thereto.

[0025] The present invention provides a composition for improving oral health comprising, as an active ingredient, a mixture of a culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and a culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

[0026] The above-mentioned oral health improvement composition may be one or more formulations selected from the group consisting of tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, gums, and candies, and may be manufactured into one selected from the group consisting of toothpaste, gargling products, gums, candies, caramels, jellies, oral sprays, oral ointments, mouthwashes, mouthwashes, and teeth whitening agents, but is not limited thereto.

[0027] A composition for improving oral health comprising a mixture of the culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and Lactobacillus plantarum GSPE05 (KCTC 15128BP) as an active ingredient may be one or more formulations selected from the group consisting of tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, gums, and candies, but is not limited thereto. Carriers, excipients, and diluents that may be included in the above probiotics include lactose, dextrose, maltodextrin, isomaltose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulating, commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, sweeteners, and acidifiers may be used.

[0028] The present invention provides a pharmaceutical composition for the treatment or prevention of respiratory diseases comprising, as an active ingredient, a mixture of culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

[0029] The above respiratory diseases may be one or more selected from the group consisting of bronchopulmonary infection, bacterial pneumonia, acute sinusitis, maxillary sinusitis, bacteremia, meningitis, chronic bronchitis, acute laryngitis, and acute otitis media.

[0030] The above pharmaceutical composition may be added in a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution in an amount preferably 0.001 to 99% by weight, more preferably 0.001 to 50% by weight, and most preferably 0.001 to 30% by weight, based on the total weight of the pharmaceutical composition.

[0031] The above pharmaceutical compositions may be formulated and used in the form of oral formulations or topical preparations, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, liquids, and aerosols, respectively, according to conventional methods. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules, and these solid formulations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition, lubricants such as magnesium stearate and talc are also used in addition to simple excipients.

[0032] The dosage of the pharmaceutical composition of the present invention may vary depending on the age, gender, and weight of the subject to treatment, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber; the determination of the dosage based on these factors is within the level of a person skilled in the art, and said dosage does not limit the scope of the present invention in any way. The pharmaceutical composition of the present invention may be administered to mammals such as rats, livestock, and humans via various routes.

[0033] The above composition of the present invention can be provided as a health functional food.

[0034] The above health functional food may be added in a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution in an amount preferably 0.001 to 99% by weight, more preferably 0.001 to 50% by weight, and most preferably 0.001 to 30% by weight, based on the total weight of the health functional food.

[0035] The above-mentioned health functional food includes forms such as tablets, capsules, powders, pills, or liquids, and foods to which the culture mixture of the present invention can be added include, for example, various types of food, beverages, chewing gum, tea, vitamin complexes, health functional food products, etc.

[0036] The present invention provides a method for preventing or treating intestinal disease in a subject in need, wherein the method comprises administering to the subject a therapeutically effective amount of a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution.

[0037] The above mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP) is a mixture obtained by mixing culture mediums of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, in a volume ratio of 1:1 to 5, and the above intestinal disease is any one of irritable bowel syndrome, ulcerative colitis, Crohn's disease, intestinal lesions associated with Behcet's disease, hemorrhagic rectal ulcer, and ileal pouchitis.

[0038] The above method can be carried out by inhibiting the activity of intestinal pathogenic microorganisms, namely Escherichiacoli, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogenes, Salmonella typhimurium, and Enterococcus faecalis, by a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution according to the present invention.

[0039] The present invention provides a method for preventing or treating oral diseases in a subject in need, wherein the method comprises administering to the subject a therapeutically effective amount of a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution.

[0040] The above mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP) is a mixture obtained by mixing culture mediums of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, in a volume ratio of 1:1 to 5, and the above oral disease may be any one of gingivitis, periodontitis, dental caries, and periapical disease.

[0041] The above method can be performed by using a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution according to the present invention to inhibit the activity of Streptococcus mutans, a dental caries bacterium, or inhibit the activity of Porphyromonas gingivalis, a causative agent of periodontal disease.

[0042] The present invention provides a method for preventing or treating respiratory diseases in a subject in need, wherein the method comprises administering to the subject a therapeutically effective amount of a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution.

[0043] The above mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP) is a mixture of culture mediums of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 cultured separately in a volume ratio of 1:1 to 5, and the above respiratory disease may be one or more selected from the group consisting of bronchopulmonary infection, bacterial pneumonia, acute sinusitis, maxillary sinusitis, bacteremia, meningitis, chronic bronchitis, acute laryngitis, and acute otitis media.

[0044] The above method can be carried out by inhibiting the activity of Moraxella catarrhalis, a respiratory bacterium, with a mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution according to the present invention.

[0045] The present invention relates to an antimicrobial composition against pathogenic bacteria comprising Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, specifically exhibiting high antimicrobial activity against one or more bacteria selected from the group consisting of Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, Streptococcus mutans, Porphyromonas gingivalis, and Moraxella catarrhalis, thereby contributing to the improvement of intestinal diseases, oral diseases, and respiratory diseases caused by these pathogenic strains.

[0046] Figure 1 is a graph showing the antibacterial effect of Bacillus cereus according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures.

[0047] Figure 2 shows the results of analyzing the inhibitory activity of Listeria monocytogenes according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures.

[0048] Figure 3 shows the results of analyzing the inhibitory activity of Salmonella typhimurium according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures.

[0049] Figure 4 shows the results of analyzing the inhibitory activity of Streptococcus mutans according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures.

[0050] Figure 5 shows the results of analyzing the inhibitory activity of Porphyromonas gingivalis according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures.

[0051] Figure 6 shows the results of analyzing the inhibitory activity of Moraxella catarrhalis according to the mixing ratio of Streptococcus salivarius G7 culture medium and Lactobacillus plantarum GSPE05 strain culture medium.

[0052] The present invention was devised after confirming that, while experimenting on the antibacterial activity of strains beneficial to the human body, the antibacterial effect against harmful bacteria is synergistically enhanced when Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 are combined.

[0053] Preferred embodiments of the present invention are described in detail below. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms, and the contents introduced herein are provided to sufficiently convey the concept of the present invention.

[0054] <Example 1. Preparation of Strain>

[0055] Streptococcus salivarius G7

[0056] The Streptococcus salivarius G7 of the present invention (accession number KCCM13161P) was isolated from the gingival tissue of a healthy Korean adult male, identified and characterized, and registered with the Korean Collection for Oral Microbiology (KCOM) (Streptococcus salivariusKCOM 2137). It was confirmed that the said strain exhibited particularly superior antibacterial, anti-inflammatory, caries-inhibiting, and bad breath-inhibiting activities compared to conventional Streptococcus salivarius, and the strain was deposited as a patent with the Korean Culture Collection Center (KCCM13161P, deposit date: April 21, 2022).

[0057] Lactobacillus plantarum GSPE05

[0058] The Lactobacillus plantarum GSPE05 strain was directly isolated from pears harvested at a farm in Naju. After confirming its superior antibacterial activity and safety compared to other lactic acid bacteria isolated together, it was identified as a new strain of Lactobacillus plantarum and was deposited as a patent (KCTC 15128BP) at the Korea Research Institute of Biotechnology on October 11, 2022.

[0059] Other strains

[0060] The strains used in the experiment were 8 types of intestinal pathogenic microorganisms, 3 types of oral harmful bacteria, and 1 type of respiratory harmful bacteria, as shown in Table 1 below, and strains obtained from each respective institution were used.

[0061] Candida albicans, Listeria monocytogenes, and Enterococcus faecalis were cultured aerobically at 30°C for 24 hours, and Porphyromonas gingivalis was cultured anaerobically at 37°C for 24 hours. In addition, the other two test bacteria and eight harmful bacteria were all cultured aerobically at 37°C for 24 hours.

[0062] No. Strain Name Strain Information Medium Type 1 Streptococcus salivarius G7 Inventive strain MRS (Difco Laboratories, Detroit, MI, USA) and TSB + 0.5% glucose (Difco Laboratories, Detroit, MI, USA) 2 Lactobacillus plantarum GSPEO 5 Inventive strain 3 Escherichiacoli KCTC 2441 Intestinal pathogenic microorganism NB (Difco Laboratories, Detroit, MI, USA) 4 Bacillus cereus KCTC 367 5 Staphylococcus aureus KCOM 133 56 Pseudomonas aeruginosa KCOM 118 27 Escherichiacoli O157:H7 ATCC 351 508 Listeria monocytogenes ATCC 191 119 Salmonella Typhimurium ATCC 140 2810 Enterococcus faecalis KCTC 5191BHI(Difco Laboratories, Detroit, MI, USA)11Streptococcus mutansKCTC 3065Dental cariogenic bacteria12Porphyromonas gingivalisKCTC 5352Causative bacteria of periodontal disease13CandidaalbicansKCTC 7122Causative bacteria of candidiasisYPD(Difco Laboratories, Detroit, MI, USA)14Moraxella catarrhalisKCCM 42706Harmful respiratory bacteriaMH(Difco Laboratories, Detroit, MI, USA)

[0063]

[0064] <Example 2. Separation of culture medium and determination of mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strains>

[0065] 5×10⁻⁶ Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strains each 7Cells were cultured under aerobic conditions at 37°C for 18 hours using TSB+0.5% glucose or MRS broth, respectively. After culture, each strain was centrifuged at 15,000 rpm for 5 minutes, and the strain and supernatant were separated. The pH of the separated supernatant was adjusted to 6.5–7.0, and the supernatant was filtered through a 0.22 μm sieve to obtain the respective culture solutions.

[0066] The culture solutions of the Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strains obtained above were prepared in a total of seven mixing ratios as shown in Table 2 and used in antimicrobial activity tests against pathogenic microorganisms and harmful bacteria related to the oral cavity and respiratory tract.

[0067] no.RatioStreptococcus salivarius G7(ml)Lactobacillus plantarum GSPE05(ml)G71:0100GSPE050:1010A1:155B1:32.57.5C1:51.78.3D5:18.31.7E3:17.52.5

[0068]

[0069] <Example 3. Evaluation of Antibacterial and Antifungal Activity>

[0070] Growth curves over 24 hours were analyzed to confirm the antimicrobial activity of 8 pathogenic microorganisms, 3 oral harmful bacteria, and 1 respiratory harmful bacterium against a culture mixture of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 mixed in the above ratio.

[0071] 12 types of harmful bacteria, each with a turbidity of 0.5 McFarland, were inoculated at 1% each into 180 μL of nutrient medium (NB medium), and then 20 μL of a mixed culture prepared with the above mixing ratio was inoculated. 200 μL of this was dispensed into 96-well plates and cultured at 37°C for 24 hours, after which the absorbance was measured at 600 nm, and the results are shown in Figures 1 to 6.

[0072] Antimicrobial activity of mixed strains against 8 types of intestinal pathogenic microorganisms

[0073] The antimicrobial activity of the culture medium of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strains mixed in the above ratio against 8 types of pathogenic microorganisms was evaluated, and it showed excellent antimicrobial effects against 3 types of pathogenic microorganisms.

[0074] Figure 1 is a graph showing the antibacterial effect of Bacillus cereus according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures. As shown in Figure 1, it was confirmed that as single-strain cultures, the Streptococcus salivarius G7 culture inhibited the activity of Bacillus cereus by 25.26%, and the Lactobacillus plantarum GSPE05 culture inhibited the activity of Bacillus cereus by 40.90%. When these two cultures were mixed, the antibacterial effect against Bacillus cereus was 66.41% for A (1:1), 77.94% for B (1:3), 91.31% for C (1:5), 71.65% for D (5:1), and 54.89% for E (3:1), with C (1:5) showing the best ability to inhibit the activity of Bacillus cereus.

[0075] Figure 2 shows the results of analyzing the inhibitory activity of Listeria monocytogenes according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strain cultures. As single-strain cultures, it was confirmed that Streptococcus salivarius G7 culture inhibited Listeria monocytogenes by 6.12%, and Lactobacillus plantarum GSPE05 culture inhibited Listeria monocytogenes by 49.80%. The antibacterial effects according to mixing ratio were A (1:1) 71.89%, B (1:3) 78.41%, C (1:5) 84.42%, D (5:1) 47.61%, and E (3:1) 57.96%, and it was confirmed that the C (1:5) group showed the best inhibition of Listeria monocytogenes activity.

[0076] Figure 3 shows the results of analyzing the inhibitory activity of Salmonella typhimurium according to the mixing ratio of culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 strains. As single strains, the Streptococcus salivarius G7 culture solution was found to be 23.90% and the Lactobacillus plantarum GSPE05 culture solution was 26.52%. The antibacterial effects according to the mixing ratio of the culture solutions were A (1:1) 54.25%, B (1:3) 54.43%, C (1:5) 69.84%, D (5:1) 47.29%, and E (3:1) 61.52%, with the C (1:5) group showing the best antibacterial effect in terms of inhibitory activity.

[0077]

[0078] Evaluation of the antibacterial and antifungal activity of a mixed strain against three types of harmful oral bacteria

[0079] As a result of evaluating the antibacterial activity against oral-related harmful bacteria, such as Streptococcus mutans and Porphyromonas gingivalis, and antifungal activity against Candida albicans of Streptococcus mutans and Porphyromonas gingivalis, of Streptococcus mutans G7 culture and Lactobacillus plantarum GSPE05 strain culture mixed in the above ratio, excellent antibacterial effects were shown against Streptococcus mutans and Porphyromonas gingivalis.

[0080] Figure 4 shows the results of analyzing the inhibitory activity of Streptococcus mutans according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 culture solutions. As single strains, it was confirmed that the Streptococcus salivarius G7 culture solution inhibited 45.16% and the Lactobacillus plantarum GSPE05 culture solution inhibited 25.73%. The antibacterial effects according to the mixing ratio of the culture solutions were A (1:1) 53.24%, B (1:3) 69.48%, C (1:5) 89.06%, D (5:1) 58.27%, and E (3:1) 54.69%, and it was confirmed that the C (1:5) group showed the best inhibition of Streptococcus mutans activity.

[0081] Figure 5 shows the results of analyzing the inhibitory activity of Porphyromonas gingivalis according to the mixing ratio of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 culture solutions. As single strains, it was confirmed that the Streptococcus salivarius G7 culture solution inhibited activity by 38.50%, and the Lactobacillus plantarum GSPE05 culture solution inhibited activity by 33.25%. The antibacterial effects according to the mixing ratio of the culture solutions were A (1:1) 74.95%, B (1:3) 55.96%, C (1:5) 84.62%, D (5:1) 66.18%, and E (3:1) 71.23%, and it was confirmed that the inhibitory activity of Porphyromonas gingivalis was most superior in the mixing ratio of the C (1:5) group.

[0082]

[0083] Antimicrobial activity of a mixed strain against one type of respiratory-related harmful bacterium

[0084] As a result of evaluating the antibacterial activity of Streptococcus salivarius G7 culture solution and Lactobacillus plantarum GSPE05 culture solution mixed in the above ratio against Moraxella catarrhalis, a respiratory-related harmful bacterium, it showed excellent antibacterial effects against Moraxella catarrhalis.

[0085]

[0086] Figure 6 shows the results of analyzing the inhibitory activity of Moraxella catarrhalis according to the mixing ratio of Streptococcus salivarius G7 culture and Lactobacillus plantarum GSPE05 strain culture. As single strains, Streptococcus salivarius G7 culture was found to inhibit 37.61% and Lactobacillus plantarum GSPE05 culture was found to inhibit 66.50%. The antibacterial effects according to mixing ratio were A (1:1) 79.03%, B (1:3) 84.66%, C (1:5) 92.54%, D (5:1) 73.42%, and E (3:1) 77.32%, and it was confirmed that the inhibitory activity of Moraxella catarrhalis was most superior at the mixing ratio of the C (1:5) group.

[0087]

[0088] The above results showed that the culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 exhibited different antibacterial activities against intestinal, oral, and respiratory harmful bacteria, respectively, and when the culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05 were mixed and applied, they showed high antibacterial activity depending on the harmful species compared to a single strain culture solution. In particular, when Streptococcus salivarius G7 culture solution and Lactobacillus plantarum GSPE05 culture solution were mixed in a 1:5 ratio (C), high antimicrobial activity was observed against 3 out of 8 pathogenic microorganisms tested (Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium), 2 out of 3 oral harmful bacteria (Streptococcus mutans, Porphyromonas gingivalis), and 1 respiratory harmful bacterium (Moraxella catarrhalis). This suggests that conditions are formed in which the antimicrobial substances contained in the culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum can act synergistically with each other.

[0089] [Consignment Number]

[0090] Depository Name: Korean Culture Collection Center (KCCM)

[0091] Trustee Number: KCCM13161P

[0092] Date of Trust: 20220421

[0093]

[0094] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC)

[0095] Trustee Number: KCTC15128BP

[0096] Date of Trust: 20221011

[0097]

Claims

1. An antimicrobial composition comprising a mixture of culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP) as an active ingredient.

2. In Paragraph 1, The above-mentioned mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution is characterized as a mixture obtained by mixing culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, in a volume ratio of 1:1 to 5, in an antimicrobial composition.

3. In Paragraph 2, The above-mentioned mixture of Streptococcus salivarius G7 (accession number KCCM13161P) culture solution and Lactobacillus plantarum GSPE05 (KCTC 15128BP) culture solution is characterized as a mixture obtained by mixing culture solutions of Streptococcus salivarius G7 and Lactobacillus plantarum GSPE05, respectively, in a volume ratio of 1:

5.

4. In Paragraph 1, An antimicrobial composition characterized by the above antimicrobial properties being antimicrobial against one or more bacteria selected from the group consisting of Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, Streptococcus mutans, Porphyromonas gingivalis, and Moraxella catarrhalis.

5. A pharmaceutical composition for the treatment or prevention of intestinal diseases comprising, as an active ingredient, a mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

6. In Paragraph 5, A pharmaceutical composition for the treatment or prevention of an intestinal disease, wherein the above intestinal disease is any one of irritable bowel syndrome, ulcerative colitis, intestinal lesions associated with Crohn's disease or Behcet's disease, hemorrhagic rectal ulcer, and ileal pouchitis.

7. A composition for improving gut health comprising a mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP) as an active ingredient.

8. A pharmaceutical composition for the treatment or prevention of oral diseases comprising, as an active ingredient, a mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

9. In Paragraph 8, A pharmaceutical composition for the treatment or prevention of oral diseases, characterized in that the above oral diseases are one or more selected from the group consisting of gingivitis, periodontitis, dental caries, and periapical disease.

10. A composition for improving oral health comprising a mixture of culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP) as an active ingredient.

11. A pharmaceutical composition for the treatment or prevention of respiratory diseases comprising, as an active ingredient, a mixture of culture medium of Streptococcus salivarius G7 (accession number KCCM13161P) and culture medium of Lactobacillus plantarum GSPE05 (KCTC 15128BP).

12. In Paragraph 11, A pharmaceutical composition for treating or preventing respiratory diseases, characterized in that the above respiratory disease is one or more selected from the group consisting of bronchopulmonary infection, bacterial pneumonia, acute sinusitis, maxillary sinusitis, bacteremia, meningitis, chronic bronchitis, acute laryngitis, and acute otitis media.

13. A composition for improving respiratory health comprising a mixture of culture solution of Streptococcus salivarius G7 (accession number KCCM13161P) and culture solution of Lactobacillus plantarum GSPE05 (KCTC 15128BP) as an active ingredient.

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