Composition for inhibiting harmful bacteria on the skin and in the oral cavity

Abstract

To provide lactic acid bacteria that are highly effective in inhibiting harmful bacteria in the skin and oral cavity.SOLUTION: A composition for inhibiting harmful bacteria in the skin and oral cavity is used for application to the skin and oral cavity, the composition containing heat-treated bacterial cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacteria HJ1 strain) as an active ingredient. Preferably, the composition has the ability to reduce odors in the skin or oral cavity to which it is applied.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This invention relates to a composition for inhibiting harmful bacteria on the skin and in the oral cavity. [Background technology]

[0002] In recent years, it has been recognized that maintaining a balance of gut microbiota is important for good health, and this is also true for the skin; it is believed that maintaining a balance of skin microbiota can help keep the skin healthy.

[0003] Regarding such technology, for example, Patent Document 1 discloses a method of using 1-ketose to promote the growth of Staphylococcus epidermidis, a known beneficial bacterium, without promoting the growth of Staphylococcus aureus and other bacteria known as harmful bacteria.

[0004] On the other hand, Patent Document 2 discloses a substance that exhibits a growth-inhibiting effect against Streptococcus mutans, which is known to cause dental caries. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Japanese Patent Publication No. 2022-184273 [Patent Document 2] Japanese Patent Publication No. 2022-27681 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Traditionally, there has been little research into using lactic acid bacteria cells to suppress harmful bacteria on the skin and in the oral cavity.

[0007] The objective of the present invention is to provide lactic acid bacteria that have excellent effects in suppressing harmful bacteria on the skin and in the oral cavity. [Means for solving the problem]

[0008] As a result of various studies conducted to achieve the above objective, the inventors of this invention discovered that Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) has an effect of suppressing harmful bacteria in the skin and oral cavity, and thus completed the present invention.

[0009] In other words, the present invention provides a composition for inhibiting harmful bacteria on the skin and in the oral cavity, which contains heat-treated cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacterium HJ1 strain (acid bacteria)) as an active ingredient and is used by applying it to the skin or oral cavity.

[0010] In the composition according to the present invention, the harmful bacteria are preferably one or more species selected from the group consisting of Staphylococcus aureus, Staphylococcus pseudintermedius, Streptococcus mutans, Porphyromonas macacae, Candida albicans, Malassezia furfur, and Helicobacter pylori.

[0011] Furthermore, it is preferable that the composition according to the present invention has an effect of reducing odor in the skin or mouth to which it is applied. [Effects of the Invention]

[0012] According to the present invention, it is possible to provide a lactic acid bacterium with excellent efficacy in suppressing harmful bacteria on the skin and in the oral cavity by utilizing the Lactobacillus HJ1 strain (Lactobacillus erythropoiesis). [Brief explanation of the drawing]

[0013] [Figure 1]This is a chart showing the results of examining the effect of Lactobacillus HJ1 strain on the growth of Staphylococcus aureus in Test Example 1. [Figure 2] This is a chart showing the results of examining the effect of Lactobacillus HJ1 strain on the growth of Staphylococcus pseudintermedius, which is the causative bacterium of impetigo, in Test Example 2. [Figure 3] This is a chart showing the results of examining the effect of Lactobacillus HJ1 strain on the growth of Streptococcus mutans, which is the causative bacterium of dental caries, in Test Example 3. [Figure 4] This is a chart showing the results of examining the effect of Lactobacillus HJ1 strain on the growth of Staphylococcus epidermis, which is a skin commensal bacterium, in Test Example 4. [Figure 5] This is a chart showing the results of examining the effect of reducing odor when Lactobacillus HJ1 strain is applied to the human oral cavity in Test Example 5. [Figure 6] This is a chart showing the results of examining the effect of reducing odor when Lactobacillus HJ1 strain is applied to the human skin in Test Example 6. [Figure 7] This is a chart showing the results of examining the effect of reducing odor when Lactobacillus HJ1 strain is applied to the oral cavity of dogs in Test Example 7. [Figure 8] This is a chart showing the results of examining the effect of reducing odor when Lactobacillus HJ1 strain is applied to the oral cavity of cats in Test Example 8. [Figure 9] This is a chart showing the results of examining the immunostimulatory ability of Lactobacillus HJ1 strain by measuring the production amount of IL-12 using mouse spleen cells in Test Example 9. [Figure 10]This figure shows the results of an investigation into the effect of Lactobacillus strain HJ1 on the growth of Porphyromonas macacae, a bacterium that causes periodontal disease, in Test Example 10. [Figure 11] This figure shows the results of an investigation into the effect of Lactobacillus strain HJ1 on the growth of Candida albicans, the causative agent of candidiasis, in Test Example 11. [Figure 12] This figure shows the results of an investigation into the effect of Lactobacillus HJ1 strain on the growth of Malassezia furfur, the causative agent of tinea versicolor and seborrheic dermatitis, in Test Example 12. [Figure 13] This figure shows the results of an investigation into the effect of Lactobacillus HJ1 strain on the growth of Helicobacter pylori, a causative agent of gastritis and peptic ulcers, in Test Example 13. [Modes for carrying out the invention]

[0014] The lactic acid bacteria used in this invention is Lactiplantibacillus plantarum, known as a plant-derived lactic acid bacterium from pickles and the like. Its scientific name is Lactiplantibacillus plantarum, and more specifically, it is Lactiplantibacillus plantarum HJ1 (accession number: NITE BP-03883), which is deposited with the Patent Microorganism Depository Center (NPMD) of the National Institute of Technology and Evaluation. Hereinafter, this lactic acid bacteria strain may be referred to as "Lactic acid bacteria strain HJ1 (Jinsankin)."

[0015] The cultivation and maintenance of Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) can be carried out by well-known methods. For example, suitable culture media include skim milk powder medium, yeast extract, peptone, meat extract, salts, and liquid media containing minerals. Commercially available media such as "MRS Broth MERCK" (trade name, Chemicals) and "Difco Lactobacilli MRS Broth" (trade name, Nippon Becton Dickinson Co., Ltd.) may also be used. Culturing can be carried out, for example, by inoculating the bacterial cells into the above-mentioned medium and then culturing them statically or by aerated stirring at conditions such as 25-35°C. For preserving live bacterial cells, they can be refrigerated while suspended in the culture medium for short periods, or frozen and stored in an antifreeze solution such as glycerol solution for long periods.

[0016] Generally, when culturing lactic acid bacteria, a decrease in pH occurs due to the metabolic products of the lactic acid bacteria (lactic acid, etc.). To avoid this, the culture may be performed while adjusting the pH by adding an alkaline agent to the culture medium (neutralization culture). As the alkaline agent to be added, for example, aqueous solutions of sodium hydroxide, potassium hydroxide, calcium hydroxide, etc., or ammonia can be used. The pH during culture may be in the range of pH 5.0 to 7.5, or in the range of pH 6.0 to 7.0, and it is preferable to adjust and maintain the pH to this range. pH adjustment may be done manually, but it is simpler and more accurate to use an automatic pH control device (pH stat). However, as for the lactic acid bacteria used in this invention, it is sufficient to obtain bacterial cells of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis), and there are no particular restrictions on how they are cultured and prepared.

[0017] When preparing Lactobacillus strain HJ1, a concentrated bacterial solution can be prepared by either concentrating the culture medium as is after cultivation, or by collecting the bacteria by means of centrifugation or filtration, further washing the bacteria with purified water, and then suspending them in purified water to a predetermined bacterial concentration. The bacterial content of Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) in 100% by mass of the concentrated bacterial solution may be in the range of 0.1 to 30% by mass, 0.5 to 20% by mass, or 1 to 10% by mass on a dry cell basis. Excipients may be included in this concentrated bacterial solution. This makes it easier to maintain the properties of the bacteria after freezing or freeze-drying, and after reconstitution with water.

[0018] Excipients are not particularly limited and include, for example, dextrin; maltodextrin; xanthan gum; sugar alcohols such as lactitol, maltitol, mannitol, sorbitol, and xylitol; sugars such as dextrose, fructose, glucose, lactose, and sucrose; and organic acids such as adipic acid, citric acid, fumaric acid, glutaric acid, malic acid, succinic acid, and tartaric acid.

[0019] In the present invention, the form of the Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) is preferably that of heat-treated cells. Heat treatment usually destroys the activity of live lactic acid bacteria, thus suppressing quality changes associated with live bacteria. Heat treatment can be carried out, for example, by autoclaving the culture solution after culturing the Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) for preparation, either directly or, if necessary, after preparing the bacterial cell concentrate as described above.

[0020] In another embodiment, the Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) may be subjected to pulverization and dispersion. The pulverization and dispersion can be carried out, for example, by pulverizing and dispersing the above-mentioned bacterial cell concentrate using means such as stirring, a mixer, a homogenizer, a ball mill, a bead mill, a jet mill, or a generator. In this case, depending on the circumstances or as needed, the re-aggregation of the resulting bacterial powder can be prevented by adding the above-mentioned excipient before the pulverization and dispersion treatment. When an excipient is included, its content may be in the range of 10 to 99% by mass, 20 to 95% by mass, or 50 to 90% by mass on a dry weight basis.

[0021] Furthermore, in another embodiment, the Lactobacillus HJ1 strain (Lactobacillus lactic acid bacteria) may be subjected to a drying and powdering treatment. Methods for drying and powdering include freeze-drying, vacuum spray drying, and spray drying using hot air. By performing spray drying using hot air (spray drying), the activity of live lactic acid bacteria is usually destroyed by the heat treatment, and dead bacterial cells can be obtained.

[0022] As can be seen in the results of the examples described later, the heat-treated bacterial cells of Lactobacillus HJ1 strain (Lactobacillus pallidum) prepared as described above do not show much inhibitory effect on the growth of Staphylococcus epidermidis, which is known as a beneficial bacterium of the skin. However, they do have an inhibitory effect on the growth of harmful bacteria such as Staphylococcus aureus, Pseudointermedius, which is the causative agent of pyoderma, Streptococcus mutans, which is the causative agent of dental caries, Porphyromonas macacae, which is the causative agent of periodontal disease, Candida fungi, which are the causative agents of candidiasis, Malassezia versicolor, which is the causative agent of tinea versicolor and seborrheic dermatitis, and Helicobacter pylori, which is the causative agent of gastritis and peptic ulcers.

[0023] Therefore, the present invention provides a composition for inhibiting harmful bacteria on the skin and in the oral cavity, which contains heat-treated cells of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) as an active ingredient and is applied to the skin or oral cavity. Furthermore, it can be used, for example, to reduce odor in the applied skin or oral cavity.

[0024] The amount of the composition according to the present invention applied to the skin or oral cavity should be appropriately determined according to the health condition and age of the subject, or the desired level of inhibitory effect on harmful bacteria. Typically, the amount applied to the application site, based on the dry weight of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis), is 0.001 mg to 100 mg / day / cm². 2 It may be within the range of 0.01 mg to 20 mg / day / cm³. 2 It may be within the range of 0.1 mg to 10 mg / day / cm³. 2 It may be within that range.

[0025] The compositions according to the present invention may be used in the form of compositions applied to the skin. That is, heat-treated bacterial cells of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) may be used as is, or combined with other raw materials or base materials for formulations to form compositions for skin use. Examples of such skin compositions include lotions, cosmetic creams, emulsions, skin milks, undermakeup, foundations, gels, jellies, powders, aerosols, bath additives, body shampoos, body rinses, soaps, cleansing foams, packs, lipsticks, lip balms, sun care products, ointments, patches, bandages, wrappers, wound dressings, etc. It should be noted that the use of such skin compositions includes not only those for humans but also those for animals.

[0026] On the other hand, the composition according to the present invention may be used in the form of an additive material to be added to such a skin composition.

[0027] The composition according to the present invention may be used in the form of a composition applied to the oral cavity. That is, heat-treated bacterial cells of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) may be used as is, or combined with other raw materials or base materials for pharmaceutical formulations to form an oral composition. Such an oral composition may be in the form of tablets, granules, lozenges, capsules, powders, drinks, jellies, gummies, candies, etc., and may be swallowed after being held in the mouth for a predetermined time. Alternatively, it may be in the form of mouthwash, chewing gum, toothpaste, etc., which are held in the mouth and not swallowed. It may also be in the form of ointments, creams, sprays, gels, lotions, etc., and these may be used for application inside the oral cavity. Note that the use of such oral compositions includes not only those for humans but also those for animals.

[0028] On the other hand, the composition according to the present invention may be used in the form of an additive material to be added to such an oral composition.

[0029] The composition according to the present invention may be used for oral treatment in the form of an animal feed product composition. That is, heat-treated bacterial cells of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) may be used as is or combined with other raw materials for animal feed products to form an animal feed product composition. Examples of animal feed products include, but are not limited to, feed for livestock, racehorses, ornamental animals, pet food, etc.

[0030] On the other hand, the composition according to the present invention may be used in the form of an additive material for animal food products, to be added to such oral animal food products.

[0031] Furthermore, if necessary, the composition according to the present invention can generally contain additive materials acceptable in cosmetics, pharmaceuticals, and foods. Examples of additive materials include surfactants, coloring agents, preservatives, thickeners, pH adjusters, emulsifiers, etc. Also, various saccharides, sweeteners, acidulants, emulsifiers, fruit juices, flavors (perfumes: peppermint oil, spearmint oil, etc.), etc. can be mentioned. More specifically, saccharides such as sucrose, glucose, fructose, honey, etc., preservatives such as glyceryl caprylate, glyceryl undecylenate, glyceryl ricinoleate, etc., thickeners such as hydrous silica, cellulose gum, xanthan gum, gellan gum, etc., pH adjusters such as sodium citrate, calcium lactate, arginine, citric acid, etc., sugar alcohols such as sorbitol, xylitol, lactitol, erythritol, palatinite, etc., emulsifiers such as lecithin, sucrose fatty acid ester, glycerol sugar fatty acid ester, etc. can be mentioned. In addition to these, various vitamins such as vitamin A, vitamin Bs, vitamin C, vitamin E, etc., herb extracts, vegetable components, milk components, cereal components, etc., cellulose, calcium stearate, modified starch, etc. can be mentioned.

[0032] In the composition according to the present invention, the content of the heat-treated cells of Lactobacillus HJ1 strain (lactic acid bacterium) may be appropriately determined in consideration of the relationship between the amount used and the effective dosage in various forms. Typically, in terms of the content in terms of the dried product of the heat-treated cells of Lactobacillus HJ1 strain (lactic acid bacterium), it may be in the range of 0.001 to 100% by mass, may be in the range of 0.005 to 50% by mass, and may be in the range of 0.001 to 10% by mass. Also, in terms of the content converted to the number of cells, it may be in the range of 2.5×10 7 ~2.5×10 12 cells / g, may be in the range of 1.25×10 8 ~1.25×10 12 cells / g, and may be in the range of 2.5×10 8 ~2.5×10 11 cells / g.

Examples

[0033] The present invention will be described in more detail below with reference to examples, but the scope of the present invention is not limited to the scope of these examples.

[0034] (Preparation Example 1) Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) was seeded in MRS liquid medium and cultured at 37°C under aeration and stirring conditions. Cultivation was stopped in the later stages of logarithmic growth, and the culture vessel was autoclaved at 121°C for 15 minutes for heat sterilization. The resulting culture solution was used in the following tests. In the figure, it is referred to as "HJ1 culture solution." The amount of bacterial cells in the culture solution was 5 × 10⁶ (dry solid content). 12 The result was cells / g.

[0035] (Preparation Example 2) The bacterial cells were recovered from the heat-sterilized culture medium obtained in Preparation Example 1 by centrifugation, suspended in purified water, and used in the following tests. In the figure, this is referred to as "HJ1 bacterial cells only." In the following tests, the amount of bacterial cells used was the same as that from the other preparation examples.

[0036] (Preparation Example 3) From the heat-sterilized culture medium obtained in Preparation Example 1, bacterial cells were recovered by centrifugation, and the same amount of dextrin as the bacterial cells was added. The mixture was then spray-dried to prepare a powder product consisting of 50% lactic acid bacteria and 50% dextrin. In the figure, this is referred to as "HJ1 product." In the following tests, the amount of bacterial cells used was the same as that used in the other preparation examples.

[0037] <Test Example 1> We investigated the effect of Lactobacillus strain HJ1 (a type of lactic acid bacterium) on the growth of Staphylococcus aureus.

[0038] Specifically, the samples prepared in Preparation Examples 1-3 were mixed in a Staphylococcus aureus growth medium (MRS medium) in a ratio of 1.25 × 10⁻⁶. 10 The culture medium was added to a concentration of cells / mL and incubated at 30°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0039] As a result, as shown in Figure 1, compared to the group cultured with Staphylococcus aureus (S. aureus) alone, the number of viable Staphylococcus aureus cells was significantly reduced in the group to which the culture solution of Lactobacillus HJ1 strain (Jinsanbacterium) prepared in Preparation Example 1 (heat-sterilized) was added to the culture medium, the group to which the bacterial cells prepared in Preparation Example 2 (including heat-sterilized Lactobacillus HJ1 strain (Jinsanbacterium)) was added to the culture medium, or the group to which the powder product prepared in Preparation Example 3 (including heat-sterilized Lactobacillus HJ1 strain (Jinsanbacterium)) was added to the culture medium.

[0040] <Test Example 2> We investigated the effect of Lactobacillus strain HJ1 (Nin-sanbacter) on the growth of Staphylococcus pseudintermedius, the causative agent of pyoderma.

[0041] Specifically, the samples prepared in Preparation Examples 1-3 were mixed in a Pseudointermedius growth medium (casein peptone + NaCl medium) at a rate of 1.25 × 10⁻⁶. 10 The culture medium was added to a concentration of cells / mL and incubated at 37°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0042] As a result, as shown in Figure 2, compared to the group cultured with Pseudointermedius alone, the number of viable Staphylococcus aureus cells was significantly reduced in the groups to which the culture solution of Lactobacillus strain HJ1 prepared in Preparation Example 1 (heat-sterilized), the bacterial cells prepared in Preparation Example 2 (containing heat-sterilized Lactobacillus strain HJ1), or the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus strain HJ1) was added to the culture medium.

[0043] <Test Example 3> We investigated the effect of Lactobacillus strain HJ1 (a type of lactic acid bacterium) on the growth of Streptococcus mutans, the causative agent of dental caries.

[0044] Specifically, the sample prepared in Preparation Example 3 was mixed with 1.25 × 10⁶ of Streptococcus mutans growth medium (Triptycase soy + yeast extract medium). 10 The culture medium was added to a concentration of cells / mL and incubated at 37°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0045] As a result, as shown in Figure 3, the number of viable *Streptococcus mutans* bacteria was significantly reduced in the group to which the powder product prepared in Preparation Example 3 (containing heat-sterilized *Lactobacillus* HJ1 strain (*Lactobacillus*)) was added to the culture medium, compared to the group to which *Streptococcus mutans* was cultured alone.

[0046] <Test Example 4> We investigated the effect of Lactobacillus strain HJ1 (a type of lactic acid bacterium) on the growth of Staphylococcus epidermis, a commensal bacterium of the skin.

[0047] Specifically, the samples prepared in Preparation Examples 2-3 were mixed in 1.25 × 10⁶ units of Staphylococcus epidermidis growth medium (MRS medium). 10 The culture medium was added to a concentration of cells / mL and incubated at 37°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0048] As a result, as shown in Figure 4, compared to the group cultured with Staphylococcus epidermis alone, no significant change in the number of viable Staphylococcus epidermis cells was observed in the groups to which the bacterial cells prepared in Preparation Example 2 (including heat-sterilized Lactobacillus HJ1 strain (Jin-san-bacter)) or the powder product prepared in Preparation Example 3 (including heat-sterilized Lactobacillus HJ1 strain) were added to the culture medium.

[0049] <Test Example 5> The odor-reducing effect of the sample prepared in Preparation Example 3 when applied to the human oral cavity was investigated.

[0050] Specifically, 8.58 × 10 10 A solution of Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) prepared with purified water to a concentration of cells / 10mL was held in the mouth, and after 5 minutes, measurement was started using a portable odor sensor ("XP-329m", Shin-Cosmos Electric Co., Ltd.). Odor measurement was performed by first drawing clean air into the sensor to return the displayed value to the base value, then inserting the sensor's intake port about 1 cm into the oral cavity while holding one's breath, and sampling the odor measurement value over time.

[0051] As a result, as shown in Figure 5, it was revealed that applying Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) to the human oral cavity has the effect of reducing odor.

[0052] <Test Example 6> The odor-reducing effect of the sample prepared in Preparation Example 3 when applied to human skin was investigated.

[0053] Specifically, 8.58 × 10 10 Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) at a concentration of cells / mL was applied to the area behind the ear of a human. After application, odor measurements were performed using a portable odor sensor ("XP-329m", Shin-Cosmos Electric Co., Ltd.). The odor measurement was performed in the same manner as in Test Example 5, except that the suction port of the sensor was placed a few millimeters above the application site.

[0054] As a result, as shown in Figure 6, it was revealed that applying Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) to human skin has the effect of reducing odor.

[0055] <Test Example 7> We investigated the odor-reducing effect of the sample prepared in Preparation Example 3 when applied to the oral cavity of a dog.

[0056] Specifically, 5 x 10 10 Cells' Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) was applied directly to the gums, and after 5 minutes, the suction port of a portable odor sensor ("XP-329m", Shin-Cosmos Electric Co., Ltd.) was fixed 1 cm from the tip of the animal's mouth. Odor measurement was performed in the same manner as in Test Example 5.

[0057] As a result, as shown in Figure 7, it was revealed that applying Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) to the oral cavity of dogs has the effect of reducing odor.

[0058] <Test Example 8> We investigated the odor-reducing effect of the sample prepared in Preparation Example 3 when applied to the oral cavity of a cat.

[0059] Specifically, 5 x 10 10 Cells' Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) was applied directly to the gums, and after 5 minutes, the suction port of a portable odor sensor ("XP-329m", Shin-Cosmos Electric Co., Ltd.) was fixed 1 cm from the tip of the animal's mouth. Odor measurement was performed in the same manner as in Test Example 5.

[0060] As a result, as shown in Figure 8, it was revealed that applying Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) to the oral cavity of cats has the effect of reducing odor.

[0061] <Test Example 9> The immune-activating ability of Lactobacillus HJ1 strain (Nin-sanbacter) was compared with other Plantarum strains in our collection by measuring IL-12 production using mouse spleen cells. Specifically, the test was conducted as follows.

[0062] Spleens were collected from BALB / cA mice (female, 10 weeks old). Cells were collected from the spleen using a cell strainer (100 μm pores), and the cell concentration was 2.5 × 10⁶. 6 Liquid culture medium RPMI-1640 (Fujifilm Wako Pure Chemical Industries) containing appropriate amounts of 10% FBS (Thermo Fisher Scientific) and penicillin / streptomycin / neomycin (PSN) antibiotic mixture (Thermo Fisher Scientific) was prepared to a cell / mL concentration. Each strain containing Lactobacillus plantarum, including Lactobacillus HJ1, was added to the cell saturation to a final concentration of 1.0 μg / mL, and the cultures were incubated at 37°C in a 5% CO2 environment for 24 hours. Subsequently, the concentrations of each cytokine in the supernatant were measured by ELISA and compared to the specific activity relative to the IL-12 production amount by OK432 (Picibanil, Chugai Pharmaceutical), an inactivated bacterial preparation used as a positive cytokine.

[0063] As a result, as shown in Figure 9, Lactobacillus HJ1 strain (Nin-sanbacter) showed higher immune-activating ability compared to other Plantarum strains in the study, in an IL-12 production measurement test using mouse spleen cells.

[0064] <Test Example 10> We investigated the effect of Lactobacillus strain HJ1 (a type of lactic acid bacterium) on the growth of Porphyromonas macacae, a bacterium that causes periodontal disease.

[0065] Specifically, the sample prepared in Preparation Example 3 was mixed with 1.4 × 10⁶ of Porphyromonas macacae growth medium (modified GAM medium 4.17% by mass, hemin 0.05% by mass, menadione 0.1% by mass, sodium carbonate 0.3% by mass, L-cysteine ​​0.14% by mass) 7 The culture medium was added to a concentration of cells / mL and incubated at 37°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0066] As a result, as shown in Figure 10, the group in which the powder product prepared in Preparation Example 3 (containing heat-sterilized lactic acid bacterium strain HJ1 (Jin-sanbachi)) was added to the culture medium showed a decreasing trend in the number of viable Porphyromonas macacae bacteria compared to the group in which Porphyromonas macacae was cultured alone.

[0067] <Test Example 11> We investigated the effect of Lactobacillus strain HJ1 (a type of lactic acid bacterium) on the growth of Candida albicans, a fungus that causes candidiasis.

[0068] Specifically, the sample prepared in Preparation Example 3 was mixed with a Candida fungal growth medium (1% glucose, 1% Tween80, 0.5% casein peptone, 0.3% yeast extract, 0.3% malt extract) in a ratio of 2.9 × 10⁶ 5 The culture medium was added to a concentration of cells / mL and incubated at 30°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0069] As a result, as shown in Figure 11, the number of viable Candida albicans fungi was significantly reduced in the group to which the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus HJ1 strain) was added to the culture medium, compared to the group to which Candida albicans was cultured alone.

[0070] <Test Example 12> We investigated the effect of Lactobacillus HJ1 strain (Lactobacillus pallidum) on the growth of Malassezia furfur, the causative agent of tinea versicolor and seborrheic dermatitis.

[0071] Specifically, the sample prepared in Preparation Example 3 was mixed with a Malassezia growth medium (3.6% by mass of malt extract, 1% by mass of Tween80, 0.6% by mass of casein peptone, 1% by mass of glycerin, and 1% by mass of olive oil) in a ratio of 3.6 × 10⁻¹⁶. 3 The culture medium was added to a concentration of cells / mL and incubated at 30°C with aeration and stirring for 24 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0072] As a result, as shown in Figure 12, the number of viable M. furfur bacteria was significantly reduced in the group to which the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus HJ1 strain) was added to the culture medium, compared to the group to which M. furfur was cultured alone.

[0073] <Test Example 13> We investigated the effect of Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) on the proliferation of Helicobacter pylori, a bacterium that causes gastritis and peptic ulcers.

[0074] Specifically, the sample prepared in Preparation Example 3 was mixed with a Helicobacter pylori growth medium (3.7% by mass of BHI medium, 10% by mass of FBS) in a ratio of 1.88 × 10⁶ 5 The culture medium was added to a concentration of cells / mL and incubated at 37°C with aeration and stirring for 72 hours. After incubation, a portion of the culture medium was spread onto an agar plate containing growth medium, and the number of viable bacteria in the culture medium after incubation was calculated from the number of colonies that developed.

[0075] As a result, as shown in Figure 13, the number of viable H. pylori bacteria was significantly reduced in the group to which the powder product prepared in Preparation Example 3 (containing heat-sterilized Lactobacillus HJ1 strain (Jin-sanbachi)) was added to the culture medium, compared to the group to which H. pylori was cultured alone.

[0076] [summary] According to the results of test examples 1-4 and 10-13, the Lactobacillus HJ1 strain (Lactobacillus erythropoiesis) did not show much inhibitory effect on the growth of Staphylococcus epidermidis, which is known as a beneficial bacterium of the skin. However, it was revealed that it has an inhibitory effect on the growth of harmful bacteria such as Staphylococcus aureus, Pseudointermedius (a causative agent of pyoderma), Streptococcus mutans (a causative agent of dental caries), Porphyromonas macacae (a causative agent of periodontal disease), Candida (a causative agent of candidiasis), Malassezia versicolor (a causative agent of tinea versicolor and seborrheic dermatitis), and Helicobacter pylori (a causative agent of gastritis and peptic ulcers).

[0077] Furthermore, the results of test examples 5-8 revealed that Lactobacillus strain HJ1 (Lactobacillus erythropoiesis) has the effect of reducing oral and skin odor.

[0078] Furthermore, according to the results of Test Example 9, Lactobacillus strain HJ1 (Nin-sanbacter) demonstrated high immune-activating ability in a test measuring IL-12 production using mouse spleen cells.

Claims

1. A microbial inhibitory composition containing heat-treated cells of Lactiplantibacillus plantarum HJ1 (lactic acid bacterium HJ1 strain) as an active ingredient, which is applied to the skin. The inhibitory composition wherein the microorganism is one or more species selected from the group consisting of Staphylococcus pseudointermedius, Candida albicans, and Malassezia furfur.

2. A microbial inhibitory composition containing heat-treated cells of Lactiplantibacillus plantarum HJ1 (Lactobacillus HJ1 strain) as an active ingredient, for use in oral cavity, The aforementioned microorganism is Porphyromonas maca. The inhibitory composition is (cae).

3. The microbial inhibitory composition according to claim 1, which has the effect of reducing the odor of the skin to which it is applied.

4. The microorganism inhibitory compound according to Claim 2, which has the effect of reducing the odor of the oral cavity to which it is applied.

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