KR102488905B1 - Novel Lactobacillus curvatus strain, and uses thereof - Google Patents

Abstract

The present invention relates to a novel lactobacillus curvatus MG5246 strain and an activity thereof. According to the present invention, the lactobacillus curvatus MG5246 strain significantly inhibited the expression of an inflammatory mediator, pro-inflammatory enzymes, cytokines, chemokines, and adhesive molecules in a cell in which a pro-inflammatory immune response is induced, inhibited the expression of genes involved in the differentiation of osteoclasts in a periodontitis animal model, and increased the expression of genes inhibiting the activity of the genes, thereby effectively improving alveolar bone loss. Therefore, the lactobacillus curvatus strain of the present invention can be utilized as an oral composition, a composition for treating periodontitis, or probiotics for the same.

Description

Novel Lactobacillus curvatus strain and uses thereof {Novel Lactobacillus curvatus strain, and uses thereof}

The present invention relates to a novel Lactobacillus curvatus MG5246 strain having anti-inflammatory, osteoclast differentiation inhibitory and osteoclast activity inhibitory effects, and uses thereof.

Oral diseases refer to all diseases that can occur in the oral cavity, including teeth. Common oral diseases include dental caries, that is, periodontal diseases such as tooth decay, periodontitis, and gingivitis. This can be. Periodontal disease includes inflammatory diseases that damage soft tissues such as the periodontal ligament and gingiva surrounding the teeth and hard tissues such as alveolar bone. play a role In the alveolar bone, the metabolism of bone formation by osteoblasts and bone resorption by osteoclasts takes place. When bone resorption exceeds bone formation for various reasons, the alveolar bone is lowered, exposing the root of the tooth, and eventually supporting the teeth. will not be able to perform This loss of alveolar bone can also be caused by gingivitis and periodontitis. Gingivitis is an inflammation localized to the soft tissue of the gums and is relatively mild and treatable. However, periodontitis is a case where gingivitis is aggravated and the inflammation progresses to the gums and around the alveolar bone, forming a periodontal pocket. It causes inflammation of the periodontal ligament and bone loss.

These oral diseases or periodontal diseases are not only the most common diseases for the adult population, both domestically and internationally, but also correspond to diseases of public health interest that require significant costs in terms of individual medical care and social management systems. According to the National Health Insurance Corporation, the number of patients with periodontal disease in Korea from 2012 to 2016 increased by about 56.6% in 4 years from 7.07 million in 2012 to 11.07 million in 2016, which corresponds to an average annual increase of 12%. Therefore, various surgical treatment methods exist and have been developed for the treatment of periodontal disease, but for individuals, surgical treatment is not only psychologically reluctant, but also carries a huge cost burden. There is a problem that continuous treatment at an appropriate time is difficult. Therefore, it can be said that not only surgical treatment but also the establishment of personal oral hygiene and prevention through it are more important for oral diseases including periodontal disease.

As a result of various studies, various strains having oral disease treatment effects have been reported. As an example, Lactobacillus gasseri SBT2055 , which shows a periodontal disease treatment effect by reducing the expression levels of IL-1β (Interleukin-1β) and TNF-α (Tumor necrosis factor-α) in mice, has been reported. In addition, Lactobacillus rhamnosus MG316 (KCTC14187BP) showing an anti-inflammatory effect and an osteoclast differentiation inhibitory effect has been reported.

On the other hand, probiotics that have a beneficial activity on the physiological activity of the host when ingested are attracting attention in various fields.

However, in order to be used as probiotics, not all strains can be used as probiotics because the strains must not have resistance to antibiotics and have no hemolytic activity.

Therefore, while having a therapeutic effect on oral diseases such as periodontitis as described above, there is a need for discovery and research on strains that can be used as probiotics.

The present inventors, while researching on new strains that can be used as probiotics while having anti-inflammatory or oral disease treatment effects, discovered new strains, their potential for use as probiotics, and positive results for anti-inflammatory and oral disease treatment. The effect was confirmed to complete the present invention.

Accordingly, an object of the present invention is to provide a novel Lactobacillus curvatus strain.

Another object of the present invention is to provide a composition for oral cavity.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating oral diseases.

Another object of the present invention is to provide a food composition for preventing or improving oral diseases.

Another object of the present invention is to provide a quasi-drug composition for preventing or improving oral diseases.

Another object of the present invention is to provide probiotics.

Another object of the present invention is to provide a feed additive composition for preventing or improving oral diseases.

In order to achieve the above object, the present invention provides a Lactobacillus curvatus MG5246 strain deposited with accession number (KCTC 14506BP).

In order to achieve the above other object, the present invention is a group consisting of a Lactobacillus curvatus MG5246 strain deposited with accession number (KCTC 14506BP), a culture thereof, an extract thereof, a fraction thereof, a dry product thereof, and a cell-free supernatant thereof Provides a composition for oral cavity containing at least one selected from.

In order to achieve the above another object, the present invention consists of the Lactobacillus curvatus MG5246 strain deposited with the accession number (KCTC 14506BP), its culture, its extract, its fraction, its dried product and its cell-free supernatant It provides a pharmaceutical composition for preventing or treating oral diseases comprising any one or more selected from the group.

In order to achieve the above another object, the present invention consists of the Lactobacillus curvatus MG5246 strain deposited with the accession number (KCTC 14506BP), its culture, its extract, its fraction, its dried product and its cell-free supernatant It provides a food composition for preventing or improving oral diseases comprising any one or more selected from the group.

In order to achieve the above another object, the present invention consists of the Lactobacillus curvatus MG5246 strain deposited with the accession number (KCTC 14506BP), its culture, its extract, its fraction, its dried product and its cell-free supernatant It provides a quasi-drug composition for preventing or improving oral diseases comprising at least one selected from the group.

In order to achieve the above another object, the present invention provides probiotics containing the Lactobacillus curvatus MG5246 strain or a culture thereof deposited under the accession number (KCTC 14506BP).

In order to achieve the above another object, the present invention consists of the Lactobacillus curvatus MG5246 strain deposited with the accession number (KCTC 14506BP), its culture, its extract, its fraction, its dried product and its cell-free supernatant It provides a composition for feed additives for preventing or improving oral diseases comprising any one or more selected from the group.

The novel Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) according to the present invention is an inflammatory mediator, proinflammatory enzyme, cytokine, chemokine and adhesion molecule in cells in which a proinflammatory immune response is induced. expression was significantly suppressed, and it was confirmed that alveolar bone loss can be effectively improved by suppressing the expression of genes involved in the differentiation of osteoclasts in periodontitis animal models and increasing the expression of genes that suppress their activity. It can be usefully utilized as a composition for treating periodontitis, or a probiotics for this.

1 is a result of confirming the effect of the strain according to the present invention on the cells in which the pro-inflammatory immune response is induced, and the effect on the production of prostaglandin E2 (PGE2) (A), cytotoxicity (B) and COX-2 (Cyclooxygenase-2) This is the result of confirming the effect on mRNA expression (C).
Figure 2 is a result of confirming the effect of the strain according to the present invention on the cells in which the pro-inflammatory immune response is induced, and the effect on TNF-α (tumor necrosis factor-α) mRNA expression (A) and IL-6 (Interleukin- 6) This is the result of confirming the effect on mRNA expression (B).
Figure 3 is a result of confirming the effect of the strain according to the present invention on the cells in which the pro-inflammatory immune response is induced, and the effect on MMP-2 (matrix metalloproteinase-2) mRNA expression (A) and MMP-8 mRNA expression This is the result of confirming the effect (B).
Figure 4 is a result of confirming the effect of the strain according to the present invention on cells in which a pro-inflammatory immune response is induced, and the effect on CXCL12 (CXC motif chemokine 12) mRNA expression (A), CXCR4 (CXC chemokine receptor type 4) mRNA This is the result of confirming the effect on expression (B) and the effect on ICAM-1 (Intercellular Adhesion Molecule-1) mRNA expression (C).
5 is a result of confirming the effect of the strain according to the present invention on alveolar bone loss in animal models, and the molar teeth of the control group (Normal), the deletion (LIgation) occurrence experimental group, and the strain administration group (MG5246) according to the present invention and morphological confirmation of alveolar bone (A) and comparison of the distance from the cemento enamel junction (CEJ) to the alveolar bone crest (ABC) of each experimental group (B).
6 is a result of confirming the effect of the strain according to the present invention on the expression of genes related to bone resorption and tissue damage in gingival tissue of an animal model, and the effect on RANKL (Receptor activator of nuclear factor kappa- Β ligand) mRNA expression ( A), the effect on OPG (osteoprotegerin) mRNA expression (B), and the RANKL/OPG ratio (C) are the results confirmed.

Hereinafter, the present invention will be described in detail.

The present invention provides a Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP).

The Lactobacillus curvatus MG5246 strain is a microorganism newly identified by the present inventors from kimchi, a fermented food, and contains a 16s rDNA sequence represented by SEQ ID NO: 1.

The inventors of the present invention deposited the Lactobacillus curvatus MG5246 strain at the Center for Biological Resources on March 22, 2021 and received the accession number KCTC 14506BP.

In the cultivation of the Lactobacillus curvatus MG5246 strain (KCTC 14506BP) according to the present invention, any medium can be used without limitation as long as it can grow the strain of the genus Lactobacillus. As an example, MRS (De Man, Rogosa and Sharpe) badges can be used.

According to one embodiment of the present invention, when the culture filtrate of the Lactobacillus curvatus MG5246 strain (KCTC 14506BP) according to the present invention was treated with cells in which a pro-inflammatory immune response was induced, an anti-inflammatory effect was exhibited.

In addition, according to one embodiment of the present invention, the culture filtrate of the Lactobacillus curvatus MG5246 strain (KCTC 14506BP) according to the present invention showed effects of inhibiting osteoclast differentiation and osteoclast activity in an animal model in which periodontitis was induced.

In addition, according to another embodiment of the present invention, the culture filtrate of the Lactobacillus curvatus MG5246 strain (KCTC 14506BP) according to the present invention is ampicillin, chloramphenicol, clindamycin, erythromycin ), Gentamycin, Kanamycin, Streptomycin, and Tetracycline, and was not resistant to one or more antibiotics selected from the group consisting of, and did not show hemolytic activity, the strain It was confirmed that it can be usefully used as a probiotic.

The present invention includes at least one selected from the group consisting of Lactobacillus curvatus MG5246 strain, a culture thereof, an extract thereof, a fraction thereof, a dried product thereof, and a cell-free supernatant thereof deposited with accession number (KCTC 14506BP) An oral composition is provided.

In the present invention, "culture medium" refers to the culture medium itself in which the strain is cultured in a suitable medium, the filtrate obtained by filtering or centrifuging the culture medium to remove the strain (filtrate, centrifuged supernatant, concentrated liquid obtained by concentrating the filtrate, culture medium) A cell disruption solution obtained by sonication or treatment with lysozyme in the culture solution, a product obtained after culturing the strain, etc. The composition of the culture solution is a component necessary for culturing a common Lactobacillus genus strain In addition, components that act synergistically on the growth of strains of the genus Lactobacillus may additionally be included, and the composition accordingly may be easily selected by those skilled in the art.

In the present invention, "Cell Free Supernatant (CFS)" refers to a pure liquid obtained by culturing a strain, centrifuging it, and removing cells filtered using a filter. The cell-free supernatant refers to a liquid obtained by culturing the strain, centrifuging it, and filtering it using a filter. The composition of the present invention may include, without limitation, the Lactobacillus curvatus MG5246 strain (KCTC 14506BP), its culture, its extract, its fraction, its concentrate, and its cell-free supernatant, and it may also be included dried. Drying methods include air drying, natural drying, spray drying and freeze drying, but are not limited thereto.

The composition for oral cavity Lactobacillus curvatus ( Lactobacillus curvatus ) Based on the anti-inflammatory effect of strain MG5246, inhibition of osteoclast differentiation and inhibition of osteoclast activity, the oral inflammatory reaction can be prevented, improved or alleviated. there is.

The oral composition may be provided in all types and formulations that can be used for oral hygiene or oral inflammatory response relief. Examples thereof include, but are not limited to, toothpaste, mouthwash, gum, oral spray, oral gel, oral ointment, and oral patch. In addition, the oral composition may be applied to oral hygiene products such as a toothbrush, floss, interdental brush, tongue cleaner, and oral tissue.

When the oral composition of the present invention is a toothpaste, Lactobacillus curvatus MG5246 strain deposited under accession number (KCTC 14506BP) as an active ingredient, its culture solution, its extract, its fraction, its dried product and its cell-free In addition to containing at least one selected from the group consisting of the supernatant, an abrasive, a binder, a moisturizer, a foaming agent, a sweetener, a whitening agent, or a flavoring agent may be further included.

The abrasive includes aluminum hydroxide, silicic anhydride, aluminum silicate, dibasic calcium phosphate dihydroxide and anhydride, tribasic calcium phosphate, calcium carbonate, calcium pyrophosphate, insoluble sodium metaphosphate, tribasic magnesium phosphate, magnesium carbonate, calcium sulfate, Polymethyl methacrylate and the like can be used alone or in combination. The content of the abrasive may be 20% by weight to 90% by weight based on the total composition, but is not limited by the content. When the oral composition is a paste composition, the binder is garagenin, various cellulose derivatives for thickening, xanthan gum, gums such as tragacanth gum, polyvinyl alcohol, sodium polyacrylate, polyacrylic acid/ Organic binders such as synthetic polymer derivatives such as maleic acid copolymers and carboxyvinyl polymers, and inorganic binders such as silica and laponite may be used alone or in combination. The content of the binder may be 0.3% to 5% by weight based on the total composition, but is not limited by the content.

In addition, the oral composition may further include sorbitol, glycerin, ethylene glycol, propylene glycol, polyether glycol, polypropylene glycol, etc. as a moisturizing agent during its preparation.

In addition, the oral composition may further include a flavoring agent or a sweetener. The sweetening agent may be sucrose, lactose, maltose, sorbitol, xylitol, sodium cyclamate, glycerin, sodium saccharin, stevioside, aspartame, etc., and the flavoring agent may be peppermint, menthol, anethol, eugenol, It may be limonene, citronenol, alpha terpineol, salicylmethyl, cineol, linalol, ethyllinalol, vanillin, thymol, spearmint oil, seji oil, rosemary oil, cinnamon oil, etc., and the sweetener or flavoring agent is alone can be used singly or in combination.

In addition, the oral composition of the present invention may include a surfactant or additional active ingredients used as a foaming component alone or in combination, and the surfactant used as the foaming component is an anionic surfactant, a nonionic surfactant, and a cationic interface. It may be any one selected from the group consisting of an active agent, and more specifically, sodium lauryl sulfate as an anionic surfactant, a copolymer of polyoxyethylene polyoxypropylene as a nonionic surfactant (poloxamer), and polyoxyethylene hydrogenated castor oil , polyoxyethylene sorbitan fatty acid esters and the like can be used without limitation.

In the case where the oral composition of the present invention is a toothpaste, Lactobacillus curvatus MG5246 strain, its culture solution, its extract, its fraction, its dried product and its acellular Except for containing at least one selected from the group consisting of supernatant, it can be prepared by a conventional toothpaste manufacturing method, and when the oral composition of the present invention is a mouthwash solution, the deposit number (KCTC 14506BP) Lactobacillus curvatus deposited as ( Lactobacillus curvatus ) MG5246 strain, its culture medium, its extract, its fraction, its dried product, and its cell-free supernatant. It can be prepared by mixing any one or more selected from the group consisting of, and formulating a mouthwash solution. Oral diseases can be prevented, improved or treated by washing the mouth 2 to 10 times a day.

In addition, when the composition for oral cavity of the present invention is a mouthwash (cleaner), it may further include a toothpaste carrier, more specifically, a non-toxic alcohol. In the present invention, at least one selected from the group consisting of the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP), its culture, its extract, its fraction, its dried product and its cell-free supernatant Silver may be included in 0.00001 to 10% by weight based on the total weight of the composition, preferably 0.0005 to 5% by weight based on the total weight of the composition, and more preferably 0.005 to 1% by weight based on the total weight of the composition. It is not limited.

In addition, the present invention includes at least one selected from the group consisting of the Lactobacillus curvatus MG5246 strain, its culture, its extract, its fraction, its dried product and its cell-free supernatant deposited under the accession number (KCTC 14506BP) To provide a pharmaceutical composition for preventing or treating oral diseases.

In the composition according to the present invention, the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) is characterized in that it inhibits or treats oral inflammation.

In addition, in the composition according to the present invention, the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) is characterized in that it inhibits the differentiation or activity of osteoclasts.

Therefore, the oral disease may be a disease caused by an inflammatory response or bone loss in the oral cavity, but is not limited thereto. More preferably, gingivitis, periodontitis, alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, and gingival atrophy. recession) and dental caries (dental caries), but may be one or more selected from the group consisting of, but is not limited thereto.

As used herein, the term "prevention" may refer to any action that inhibits or delays the onset of oral diseases by administering the composition for preventing or treating oral diseases according to the present invention to a subject.

As used herein, the term "treatment" refers to an approach for obtaining a beneficial or desirable clinical result, whether or not a beneficial or desirable clinical result is detectable or not possible, and whether partial or total, for the purposes of the present invention. alleviation of symptoms, reduction of severity of disease, stabilization (i.e., no worsening) of disease, slowing or slowing of disease progression, amelioration or palliation and relief of disease state, but are not limited to . Thus, treatment refers to both therapeutic treatment and prophylactic treatment, and those requiring treatment include conditions in which a disease is to be prevented as well as conditions in which a disease is already present. In addition, it may mean any action that improves or benefits the symptoms of oral diseases by administering the pharmaceutical composition of the present invention to a subject.

The pharmaceutical composition of the present invention may be administered to an “individual” who has or is likely to develop an oral disease, and the “individual” may mean all animals, including humans.

The pharmaceutical composition of the present invention can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. there is. Carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are any selected from the group consisting of Lactobacillus rhamnosus, its culture medium, its extract, and its cell-free supernatant of the present invention. It is prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with a mixture of one or more collagen hydrolysates. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspensions. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The dosage of the pharmaceutical composition of the present invention will vary depending on the age, sex, and weight of the subject to be treated, 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. Determination of dosage based on these factors is within the level of those skilled in the art, and generally dosages range from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 0.1 mg/kg/day to 1000 mg/kg/day. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, and humans through various routes, and can be used both parenterally and orally, but is preferably administered parenterally.

Forms for parenteral administration include toothpastes, mouthwashes, topical agents (creams, ointments, dressing solutions, sprays, other coating agents, etc.). As an example of the formulation of the topical administration, the pharmaceutical composition for preventing or treating oral diseases according to the present invention is fixed to a film or patch containing a water-soluble polymer and formulated so that it can be attached to teeth and surrounding areas. it could be

In the present invention, the pharmaceutical composition for preventing or treating oral diseases is Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) as an active ingredient, its culture solution, its extract, its fraction, its In addition to at least one selected from the group consisting of dry matter and acellular supernatant thereof, any compound or natural extract known to have a therapeutic effect on oral diseases and whose safety has already been verified to increase the therapeutic effect of oral diseases may be further included. there is.

In addition, the pharmaceutical composition for preventing or treating oral diseases of the present invention may be used in conjunction with surgical treatment of oral diseases.

In addition, the present invention includes at least one selected from the group consisting of a Lactobacillus curvatus MG5246 strain, a culture thereof, an extract thereof, a fraction thereof, a dried product thereof, and a cell-free supernatant thereof deposited under accession number (KCTC 14506BP) To provide a food composition for preventing or improving oral diseases.

In addition, the present invention provides probiotics, including the Lactobacillus curvatus MG5246 strain or its culture medium deposited with accession number (KCTC 14506BP).

Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) included in the probiotics of the present invention has no antibiotic resistance, no hemolytic activity, anti-inflammatory effect, inhibition of osteoclast differentiation and osteoclast Since it shows an activity inhibitory effect, the probiotics of the present invention can be used for preventing and improving oral diseases.

The food composition or probiotics according to the present invention can be used in all forms such as functional food, nutritional supplement, health food, health functional food and food additives. include

In the present invention, the term "health functional food" refers to a food manufactured and processed by using a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplement. It refers to food designed and processed so that it can sufficiently exert biological control functions such as biological defense, regulation of circadian rhythm, prevention and recovery of disease, etc. say what you can

Food compositions or probiotics according to the present invention can be prepared in various forms according to conventional methods known in the art.

For example, as a health food, the group consisting of the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) of the present invention, its culture, its extract, its fraction, its dried product and its cell-free supernatant Any one or more selected from may be granulated, encapsulated, and powdered to be ingested or prepared in the form of tea, juice, and drink to be consumed. In addition, at least one selected from the group consisting of the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) of the present invention, its culture, its extract, its fraction, its dried product and its cell-free supernatant It can be prepared in the form of a composition by mixing with a known substance or active ingredient known to have an effect of preventing, improving or treating oral diseases.

In addition, functional foods include beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruit, bottled food, jam, marmalade, etc.), fish, meat, and their processed foods (e.g., ham, sausage corned beef, etc.) , breads and noodles (e.g. udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine, vegetable protein, retort Food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.) deposited under the accession number (KCTC 14506BP) of the present invention, etc. Lactobacillus curvatus MG5246 strain, its culture medium, its extract, its fraction, It can be prepared by adding at least one selected from the group consisting of its dried product and its cell-free supernatant.

Among the food compositions of the present invention, in the group consisting of the Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) of the present invention, its culture, its extract, its fraction, its dried product and its cell-free supernatant The preferred content of one or more selected ones is not limited thereto, but may be, for example, 0.01 to 80% by weight of the finally prepared food, preferably 0.01 to 50% by weight of the finally prepared food.

In addition, in order to use the food composition of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or concentrate.

In addition, the present invention includes at least one selected from the group consisting of a Lactobacillus curvatus MG5246 strain, a culture thereof, an extract thereof, a fraction thereof, a dried product thereof, and a cell-free supernatant thereof deposited under accession number (KCTC 14506BP) To provide a quasi-drug composition for preventing or improving oral diseases.

In the present invention, the "quasi-drug" refers to products that have a milder effect than pharmaceuticals among products used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing human or animal diseases, for example, according to the Pharmaceutical Affairs Act. According to this, quasi-drugs are items excluding items used for pharmaceutical purposes, and include products used for the treatment or prevention of human and/or animal diseases, and products with minor or no direct action on the human body.

Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) of the present invention, a culture thereof, an extract thereof, a fraction thereof, a dried product thereof, and a cell-free supernatant thereof. When used as an additive, it may be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and may be appropriately used according to conventional methods. The mixing ratio with other components used together may be appropriately determined depending on the purpose of use (eg, prevention, improvement or therapeutic treatment).

In the present invention, the quasi-drug composition may be formulated into ointments, creams, gels, lotions, liquids, dressings, patches, blisters, tapes, mists, external powders, and sprays.

The present invention includes at least one selected from the group consisting of Lactobacillus curvatus MG5246 strain, a culture thereof, an extract thereof, a fraction thereof, a dried product thereof, and a cell-free supernatant thereof deposited with accession number (KCTC 14506BP) , It provides a composition for feed additives for preventing or improving oral diseases.

The term "feed" of the present invention means any natural or artificial oral diet, one meal, etc., or a component of the one meal meal eaten by animals, and the composition for adding feed for preventing or improving oral diseases according to the present invention is effective The feed included as a component can be prepared with various types of feed known in the art, and preferably may include concentrated feed, roughage and/or special feed, but is not limited thereto.

In the present invention, the term "composition for feed additive" may mean "feed additive", which alleviates symptoms of disease in animals, supplements nutrients and prevents weight loss, enhances the digestibility of fiber in feed, improves milk quality, and breeds. It includes substances added to feed for the purpose of various effects, such as preventing disorders, improving conception rates, and preventing high-temperature stress in the summer.

The composition for feed additive of the present invention corresponds to supplementary feed under the Feed Management Act, and is a mineral preparation such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, and a trace mineral preparation such as zinc, copper, cobalt, and selenium. , vitamins such as kerotene, vitamins A, D, E, nicotinic acid, and vitamin B complex, protected amino acids such as methionine and lysine, protected fatty acids such as calcium salts of fatty acids, probiotics (lactic acid bacteria), yeast cultures, and fungal fermentation products. Probiotics, yeasts, and the like may be further included.

Among the feeds, the enriched feed includes seed fruits including grains such as wheat, oats and corn, bran including rice bran, bran, barley bran, etc. as a by-product obtained by refining grains, soybeans, fluids, sesame seeds, linseed, coco Sesame cakes, which are by-products obtained from oil extraction of palms, and residual starches, which are the main component of starch residues, which are the remainder after removing starch from sweet potatoes and potatoes, fish meal, fish residues, and fresh liquid obtained from fish Animal substances such as concentrated fish soluble, meat meal, blood meal, feather meal, skim milk powder, dried whey, which is the balance when manufacturing cheese from milk and casein from skim milk, dried Feed, yeast, chlorella, algae, but not limited thereto.

Forages among the feed include raw grass feed such as wild grass, grass, green cutting, turnip for feed, beet for feed, root vegetables such as Lutherbearer, a kind of turnip, raw grass, green crops, grain Silage, which is a stored feed made by lactic acid fermentation by filling the back into a silo, wild grass, hay dried by cutting grass, straw of breeding crops, and leaves of legumes, but is not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that are likely to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storability of feeds. There are feed additives and dietary supplements, which are substances added in small amounts, but are not limited thereto.

The feed additive composition according to the present invention is Lactobacillus curvatus MG5246 strain, its culture medium, its extract, its fraction, its dried product and its extract in an appropriate effective concentration range according to various feed manufacturing methods known in the art It can be prepared by adding at least one selected from the group consisting of cell-free supernatant.

The feed additive according to the present invention is effective against gingivitis, periodontitis, alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, gums Any object for the purpose of preventing or improving oral diseases such as gingival recession or dental caries can be applied without limitation. For example, it can be applied to any object such as non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds and fish.

The contents of the present invention described above are equally applied to each other unless contradictory to each other, and implementation by adding appropriate changes by a person skilled in the art is also included in the scope of the present invention.

Hereinafter, the present invention will be described in detail through examples, but the scope of the present invention is not limited only to the following examples.

Experimental Example 1. Material and cell preparation

Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were purchased from WelGENE Inc. (Daegu, Korea). Prostaglandin E2 (PGE2) enzyme-linked immunosorbent assay kit was purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). Taqman ^® Universal master mix and Taqman ^® probes (50-fluorescein based reporter dye; 30-TAMRA quencher and high capacity RNA-to-cDNA kit) are from Applied Bio Systems was purchased from Applied Biosystems (Foster City, CA, USA). In addition, Pepsin, pancreatin, and other reagents were purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA). USA))).

HGF-1 cells (purchased from the American Type Culture Collection (ATCC), (Manassas, VA, USA)) were cultured in DMEM containing 10% FBS (v/v) and 1% penicillin-streptomycin (100 units/mL). 37 ℃, 5% CO ₂ It was cultured under conditions.

The medium was replaced every 2-3 days, and subculture was performed when the bottom of the plate was full of cells. Only cells that had been subcultured between 4 and 10 were used, and cytotoxicity was confirmed using MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. did

Experimental Example 2. Confirmation of the characteristics of the strain according to the present invention

2-1. Check the survival rate in the stomach

In order to evaluate the gastrointestinal resistance of the strain ( L. curvatus MG5246) according to the present invention, MRS (De Man, Rogosa and Sharpe) medium (1% peptone, 0.8% meat extract, 0.4% yeast extract, 2% glucose, 0.5% sodium acetate, 0.2% dipotassium hydrogen phosphate, 0.02% magnesium sulfate heptahydrate, 0.005% After incubation at 37 ° C. for 24 hours with manganese sulfate tetrahydrate and 0.02% triammonium citrate, strains were harvested by centrifugation (3460 xg, 10 minutes). The pellets were washed twice with phosphate buffered saline (PBS) at pH 7 and then resuspended in simulated gastric fluid (SGF (PBS with 1 g/L pepsin, pH 3 and pH 4)) ( 10 ⁷ CFU/ml). In addition, it was resuspended in simulated intestinal fluid (SIF (PBS containing 1 g/L of pancreatin, pH 7 and pH 8)) (10 ⁷ CFU/ml).

After culturing at 37° C. for 3 to 4 hours, viable cells were confirmed using MRS agar medium. Strain survival was confirmed through Equation 1 below.

[Equation 1]

2-2. Antibiotic Resistance Check

The antibiotic resistance of the strain according to the present invention ( L. curvatus MG5246) was analyzed through minimum inhibitory concentration (MIC) test strip analysis. After culturing in MRS medium at 37 ° C for 18 hours, the obtained strain was centrifuged (3460 xg, 10 minutes), washed twice with PBS (pH 7), and resuspended in PBS to obtain 0.5 McFarland turbidity. obtained.

The resuspended solution was cultured in LSM medium containing 1.5% agar (a mixture of 90% Iso-Sensitest (OXOID, Hampshire, UK) and 10% MRS medium). Plates were dried for 10 minutes and 8 antibiotics (Ampicillin, Chloramphenicol, Clindamycin, erythromycin, Gentamycin, Kanamycin, Streptomycin or tetra MIC test strips (Liofilchem, Italy) for cycline (Tetracycline) were placed on the medium surface according to the manufacturer's instructions. After incubating it at 37 ° C. for 24 hours, the results were confirmed. Antibiotic susceptibility was determined according to the guidelines of the European Food Safety Authority (EFSA).

2-3. Confirmation of hemolytic properties

In order to evaluate the hemolytic activity, the strain according to the present invention ( L. curvatus MG5246) was cultured in MRS medium at 37 ° C. for 18 hours, and then TSA containing 5% sheep blood (MB CELL, Seoul, Korea) It was smeared on tryptic soy agar) medium (Difco, USA), incubated for 48 hours under conditions of 37° C. and 10 CO ₂ , and then the hemolytic activity was confirmed based on lysis of red blood cells around the generated colonies.

Experimental Example 3. Confirmation of the effect on PGE2 production

HGF-1 cells were seeded in a 6 cm plate at a density of 4×10 ⁵ cells/well and cultured until 90% of the bottom of the plate was occupied.

The cells were treated with 5 μg/ml of Pg-LPS (Invivogen, USA) in a fresh phenol-free DMEM medium to stimulate an immune response, and the cell-free supernatant (cell free) of the strain according to the present invention ( L. curvatus MG5246) supernatant, CFS) at various concentrations (100, 200 and 400 LG/ml) and then cultured for 24 hours. After 24 hours had elapsed, the production of PGE2 was confirmed from the culture supernatant.

Experimental Example 4. Periodontitis induced animal model production

Male rats (Sprague-Dawley rats, (Orient Bio, Korea)), 6 weeks old, weighing 400 to 430 g were purchased and subjected to a 12-hour light-dark cycle, 23 ± 3 ° C, 55 ± 15% humidity kept as In addition, purified water and standard diet were fed ad libitum.

After a one-week adaptation period, the rats were randomly divided into three groups, with 10 rats per experimental group [(1): no ligation, (2) ligation control group, (3) ligation and according to the present invention Strain ( L. curvatus MG5246) administration (2×10 ⁸ CFU/day)].

Current Lactobacillus strains do not cause toxic reactions at a concentration of 1×10 ¹⁰ CUF/kg/day, and most of the doses of probiotics administered to humans are 10 ⁸ , up to 10 ⁹ CFU (2×10 ⁸ The dosage was determined by CFU/kg/day). Rats in each experimental group were sacrificed after 8 weeks, and gingival tissues were obtained for further analysis.

This experiment was conducted with the approval of the Institutional Animal Care and Use Committee (KNOTUS 21-KE-032).

Experimental Example 5. Micro-computerized tomography (Micro-CT) analysis

After 8 weeks, morphological analysis of ligated molar teeth and alveolar bone of rats in each experimental group was performed using a CT 80 (SCANCO Medical, Switzerland). The distance from the cemento enamel junction (CEJ) to the alveolar bone crest (ABC) was used as an indicator of alveolar bone loss.

Experimental Example 6. RNA extraction and qRT-PCR (quantitative real time polymerase chain reaction) analysis

qRT-PCR analysis was performed according to a known method using HGF-1 cell lysate and rat gingival tissue. At this time, the target genes [COX-2 (cyclooxygenase-2), TNF-α (tumor necrosis factor-α), IL-6 (Interleukin-6), MMP-2 (matrix metalloproteinase-2), MMP-8, CXCL12 (C-X-C Motif chemokine 12), CXCR4 (C-X-C chemokine receptor type 4), ICAM-1 (Intercellular Adhesion Molecule-1), RANKL (Receptor activator of nuclear factor kappa-Β ligand), and OPG (osteoprotegerin)] expression was measured using the StepOne Plus RTPCR system. (Applied Biosystems, USA).

Relative changes in target gene expression levels were normalized to β-actin values.

To this end, ThermoFisher Co. (USA), the following primers were used: β-actin (Mm00607939_s1 and Rn00667869_m1), TNF-α (Mm00437214_m1), MMP-2 (Rn01538170_m1), RANKL (Rn00589289_m1), OPG (Rn00563499-6_m1), IL Mm00446190_m1) COX-2 (Hs00153133_m1), CXCL12 (Hs03676656_m1), CXCR4 (Hs00607978_s1), ICAM1 (Hs00164932_m1).

Experimental Example 7. Statistical Analysis

All cell culture experiments and analyzes were performed in triplicate, and data were presented as mean ± SD (mean ± SD). One-way analysis of variance and Duncan's multiple comparison test (SPSS 25; SPSS, Inc., Chicago, IL, USA) were performed to confirm significant differences in each experimental group. (p/0.05).

Example 1. Lactobacillus Curvatus MG5246 ( Lactobacillus curvatus MG5246, L. curvatus Isolation and culture of MG5246)

L. curvatus MG5246 of the present invention was isolated from kimchi, a fermented food. More specifically, after pulverizing the kimchi sample, 10 g was mixed with 90 mL of a 0.85% saline solution and stirred for 5 minutes using a stomacher (simple mixer, 3M). 1mL of the mixed solution was diluted with 0.85% saline diluent to 10 ⁸ in decimal. Each diluted sample was spread on MRS agar and Rogosa agar medium and then incubated at 37° C. for 24 to 48 hours. The formed colony was inoculated into MRS medium to which BCP (Bromocresol purple) was added to confirm whether a yellow ring was formed, and then Gram staining (positive), biochemical reaction (catalase negative), and morphology (rod) of the colony forming a yellow ring were checked. ) was confirmed, and the bacterial species was confirmed through 16S rRNA. Identified strains were stored at -80°C as 20% glycerol stock.

L. curvatus MG5246 of the present invention contains the 16S rRNA gene represented by the nucleotide sequence of SEQ ID NO: 1. In addition, the L. curvatus MG5246 was patented at the Center for Biological Resources on March 22, 2021, and was given accession number KCTC 14506BP.

The L. curvatus MG5246 strain was aerobically cultured in MRS medium at 37° C. for 24 hours. In addition, the supernatant was obtained by centrifugation (3460 xg, 10 minutes). The obtained supernatant was filtered using a 0.2 μm polytetrafluoroethylene (PTFE) membrane filter (ADVANTEC, Tokyo, Japan) to prepare a cell free supernatant (CFS), which was lyophilized.

Example 2. Lactobacillus Curvatus MG5246 ( Lactobacillus curvatus MG5246, L. curvatus MG5246) characterization

2-1. Check the survival rate in the stomach

In the utilization of probiotics, survival in the gastrointestinal tract is very important. Therefore, the gastrointestinal resistance of the strain ( L. curvatus MG5246) according to the present invention was confirmed and shown in Table 1 below.

strain Initial counts Simulated gastric fluid (SGF)
(survival rate %) Simulated intestinal fluid (SIF)
(survival rate %) pH 3 pH 4 pH 7 pH 8 Lactobacillus curvatus MG5246 6.89 ±0.27 5.65 ±0.05
(82.00%) 6.68 ±0.05
(96.95%) 6.80 ±0.00
(98.69%) 6.79 ±0.01
(98.55%)

As confirmed in Table 1, the L. curvatus MG5246 strain according to the present invention showed a survival rate of 82% compared to the initial (initial count) in simulated gastric fluid at pH 3. In addition, the L. curvatus MG5246 strain showed a viable cell count of >6 log CFU/ml and a viability of >96% in an environment of pH 4-8. In general, the survival rate of probiotic strains is 70% to 103% in SGF. Therefore, it was confirmed that the L. curvatus MG5246 strain according to the present invention had viability acceptable for use as a probiotic strain.

2-2. Confirmation of antibiotic resistance and hemolytic activity

Antibiotic resistance and hemolysis are very important in the safety of using probiotic strains. Therefore, the MIC and hemolytic activity of the L. curvatus MG5246 strain against 8 antibiotics according to the present invention were confirmed.

As shown in Table 2, the 8 antibiotics of the L. curvatus MG5246 strain according to the present invention showed values significantly lower than the cut-off MIC values of the EFSA guidelines. In addition, the L. curvatus MG5246 strain according to the present invention did not show hemolytic activity. Therefore, it was confirmed that the L. curvatus MG5246 strain according to the present invention did not cause safety problems due to antibiotic resistance and hemolytic activity.

Example 3. Confirmation of the effect of CFS on PGE2 production and inflammatory gene expression in HGF-1 cells stimulated with Pg-LPS

PGE2 plays an important role in the progress of the inflammatory response and is involved in the induction of pro-inflammatory immune responses such as cytokine secretion, extracellular matrix degradation and alveolar bone loss.

In addition, TNF-α is a primary inflammatory mediator and promotes the induction of secondary mediators such as chemokines and COX-2. COX-2 expression and PGE2 production are significantly up-regulated in inflamed gingival tissues, and it is known that IL-1β and periodontal bacterial components increase PGE2 production.

In addition, as periodontitis progresses, collagen degradation is increased by MMPs secreted from gingival fibroblasts. It was confirmed that the fibroblasts secrete multiple MMPs, and the expression level of MMP-2 was increased in the gingival tissue of chronic adult periodontitis. In addition, MMP-8 is a major collagen degrading protease and is known to be closely related to the severity of periodontal inflammation. Therefore, inhibition of MMP-2 and MMP-8 may delay the progression or severity of periodontal disease.

Accordingly, the effects of L. curvatus MG5246 CFS according to the present invention on PGE2 production and inflammatory gene expression in HGF-1 cells stimulated with pro-inflammatory immune response by Pg-LPS were confirmed. As a result, as shown in (A) in FIG. 1, it was confirmed that the production of PGE2 was remarkably reduced by the CFS treatment at 200 μg/ml or 400 μg/ml. In addition, as shown in (B) of FIG. 1, cytotoxicity was not confirmed at all concentrations of CFS tested. Also, as shown in (C) of FIG. 1, COX-2 gene expression was significantly reduced by CFS treatment at 200 μg/ml or 400 μg/ml.

In addition, the expression of TNF-α was significantly reduced (up to 60% by 200 μg/ml CFS (p<0.05)), as shown in (A) in FIG. 2, and as shown in (B) in FIG. The expression of IL-6, one of the pro-inflammatory cytokines, was also significantly inhibited by 200 and 400 μg/ml CFS (p<0.05). In addition, as shown in (A) and (B) in FIG. 3, it was confirmed that the gene expression of MMP-2 and MMP-8 was effectively down-regulated by 100 μg/ml CFS treatment (p<0.05).

Example 4. Confirmation of the effect of CFS on chemokine gene expression in HGF-1 cells stimulated with Pg-LPS

Chemokines are cytokines that promote the migration of immune cells through specific receptors. Among chemokines, CXCL12 shows increased expression in patients with rheumatoid arthritis and osteoarthritis, and is known to be closely related to bone resorption by osteoclasts. In addition, ICAM-1 is an inflammatory protein involved in the adhesion and migration of inflammatory cells.

Accordingly, the effect of L. curvatus MG5246 CFS according to the present invention on chemokine gene expression in cells in which a pro-inflammatory immune response was induced was confirmed by qRT-PCR. As a result, as shown in (A) and (B) in FIG. 4, the gene expression of CXCL12 and its receptor, CXCR4, was reduced by 42% and 70%, respectively, by 400 μlg/mL L. curvatus MG5246 CFS treatment according to the present invention. did. Also, as shown in (C) of FIG. 4, L. curvatus MG5246 CFS according to the present invention decreased ICAM-1 gene expression in a concentration-dependent manner.

Therefore, it was confirmed that the L. curvatus MG5246 CFS according to the present invention inhibits the expression of CXCL12, CXCR4 and ICAM-1, and thus exhibits periodontal inflammation relieving and therapeutic effects.

Example 5. Confirmation of the effect on alveolar bone loss in periodontitis animal models

The L. curvatus MG5246 strain according to the present invention was fed to an animal model inducing periodontitis, and the effect on alveolar bone loss was confirmed, and the results are shown in FIG. 5 .

As a result, as shown in (A) and (B) in FIG. 5, although the distance of CEJ-ABC was significantly increased in the ligation control group, in the case of the experimental group fed the L. curvatus MG5246 strain according to the present invention, the CEJ-ABC It was confirmed that the distance was significantly reduced.

In addition, the effect of the L. curvatus MG5246 strain according to the present invention on the expression of genes related to bone resorption and tissue damage in gingival tissue was confirmed through qRT-PCR. As a result, as shown in (A) and (B) in FIG. 6, the L. curvatus MG5246 strain-administered experimental group significantly reduced the expression of RANKL, which induces the differentiation of osteoclasts, while OPG, which inhibits the activity of osteoclasts, The expression of was increased 2.2 times compared to the ligation control group. In addition, the RANKL / OPG ratio was confirmed, and as shown in (C) in FIG. 6, it was confirmed that the RANKL / OPG ratio significantly increased by ligation was significantly reduced by administration of the L. curvatus MG5246 strain according to the present invention.

Overall, the novel Lactobacillus curvatus MG5246 ( Lactobacillus curvatus MG5246, L. curvatus MG5246) according to the present invention has no cytotoxicity, antibiotic resistance and hemolytic activity, so its safety in use as a probiotic was confirmed. In addition, inflammatory mediators (PGE2), pro-inflammatory enzymes (COX-2 and MMP), cytokines (TNF-a and IL-6), chemokines (CXCL12 and CXCR4) and adhesion molecules (ICAM -1) was significantly suppressed, and it was confirmed that alveolar bone loss can be effectively improved by suppressing the expression of a gene involved in the differentiation of osteoclasts in a periodontitis animal model and increasing the expression of a gene that suppresses its activity. Bar, it can be usefully utilized as a composition for oral cavity, a composition for treating periodontitis, or probiotics for this.

Biological Resource Center KCTC14506BP 20210322

<110> MEDIOGEN Co.,Ltd. <120> Novel Lactobacillus curvatus strain, and uses thereof <130> 1-49P <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1437 <212> DNA <213> artificial sequence <220> <223> Lactobacillus curvatus MG5246 16S rRNA <400> 1 atgcagtcga acgcactctc gttagattga agaagcttgc ttctgattga taacatttga 60 gtgagtggcg gacgggtgag taacacgtgg gtaacctgcc ctaaagtggg ggataacatt 120 tggaaacaga tgctaatacc gcataaaacc tagcaccgca tggtgcaagg ttgaaagatg 180 gtttcggcta tcactttagg atggacccgc ggtgcattag ttagttggtg aggtaaaggc 240 tcaccaagac cgtgatgcat agccgacctg agagggtaat cggccacact gggactgaga 300 cacggcccag actcctacgg gaggcagcag tagggaatct tccacaatgg acgaaagtct 360 gatggagcaa cgccgcgtga gtgaagaagg ttttcggatc gtaaaactct gttgttggag 420 aagaacgtat ttgatagtaa ctgatcaggt agtgacggta tccaacga aagccacggc 480 taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gatttattgg 540 gcgtaaagcg agcgcaggcg gtttcttaag tctgatgtga aagccttcgg ctcaaccgaa 600 gaagtgcatc ggaaactggg aaacttgagt gcagaagagg acagtggaac tccatgtgta 660 gcggtgaaat gcgtagatat atggaagaac accagtggcg aaggcggctg tctggtctgt 720 aactgacgct gaggctcgaa agcatgggta gcaaacagga ttagataccc tggtagtcca 780 tgccgtaaac gatgagtgct aggtgttgga gggtttccgc ccttcagtgc cgcagctaac 840 gcattaagca ctccgcctgg ggagtacgac cgcaaggttg aaactcaaag gaattgacgg 900 gggcccgcac aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca 960 ggtcttgaca tcctttgacc actctagaga tagagctttc ccttcgggga caaagtgaca 1020 ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080 cgcaaccctt attactagtt gccagcattt agttgggcac tctagtgaga ctgccggtga 1140 caaaccggag gaaggtgggg acgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200 cacgtgctac aatggatggt acaacgagtc gcgagaccgc gaggtttagc taatctctta 1260 aaaccattct cagttcggat tgtaggctgc aactcgccta catgaagccg gaatcgctag 1320 taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380 acaccatgag agtttgtaac acccaaagcc ggtgaggtaa ccttcggggag ccagccg 1437

Claims (15)

Lactobacillus curvatus MG5246 strain deposited under accession number (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1.
delete According to claim 1,
The strain is characterized in that it has one or more effects selected from the group consisting of anti-inflammatory, inhibition of osteoclast differentiation and inhibition of osteoclast activity, the strain.
According to claim 1,
The strain is from the group consisting of Ampicillin, Chloramphenicol, Clindamycin, Erythromycin, Gentamycin, Kanamycin, Streptomycin and Tetracycline A strain, characterized in that it does not have resistance to any one or more selected antibiotics.
Consisting of the Lactobacillus curvatus MG5246 strain deposited under Accession No. (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1, its culture, its extract, its fraction, its dried product and its acellular supernatant containing at least one selected from the group
Gingivitis, periodontitis, alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia, gingival recession and teeth An oral composition for preventing or improving oral inflammation caused by one or more selected from the group consisting of dental caries.
delete Consisting of the Lactobacillus curvatus MG5246 strain deposited under Accession No. (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1, its culture, its extract, its fraction, its dried product and its acellular supernatant Including any one or more selected from the group, gingivitis, periodontitis, alveolar bone breakage, alveolar bone osteoporosis, alveolar bone osteomalacia, alveolar bone osteopenia ), a pharmaceutical composition for preventing or treating one or more oral diseases selected from the group consisting of gingival recession and dental caries.
delete According to claim 7,
Lactobacillus curvatus MG5246 strain deposited with the accession number (KCTC 14506BP) is a pharmaceutical composition, characterized in that for inhibiting or treating inflammation in the oral cavity.
According to claim 7,
Lactobacillus curvatus MG5246 strain deposited under the accession number (KCTC 14506BP) is a pharmaceutical composition, characterized in that inhibits the differentiation or activity of osteoclasts.
Consisting of the Lactobacillus curvatus MG5246 strain deposited under Accession No. (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1, its culture, its extract, its fraction, its dried product and its acellular supernatant A food composition for preventing or improving oral diseases comprising at least one selected from the group.
Consisting of the Lactobacillus curvatus MG5246 strain deposited under Accession No. (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1, its culture, its extract, its fraction, its dried product and its acellular supernatant A quasi-drug composition for preventing or improving oral diseases comprising at least one selected from the group.
Lactobacillus curvatus deposited with accession number (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1 ( Lactobacillus curvatus ) MG5246 strain or probiotics containing a culture thereof.
According to claim 13,
Characterized in that the probiotics are used for preventing and improving oral diseases, probiotics.
Consisting of the Lactobacillus curvatus MG5246 strain deposited under Accession No. (KCTC 14506BP) containing the 16s rDNA sequence represented by SEQ ID NO: 1, its culture, its extract, its fraction, its dried product and its acellular supernatant Feed additive composition for preventing or improving oral disease, comprising any one or more selected from the group.

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