KR102491641B1 - Lacticaseibacillus paracasei strain for suppressing periodontal disease inducing bacteria and use thereof - Google Patents

Abstract

The present invention provides a Lacticaseibacillus paracasei strain having an activity to inhibit periodontal disease-causing bacteria, and a use thereof. The strain is a Lacticaseibacillus paracasei subsp. tolerans LMT18-32 (accession number: KCTC14823BP) strain having an activity to inhibit periodontal disease-causing bacteria, and the periodontal disease-causing bacteria are Porphyromonas gingivalis.

Description

Lacticaseibacillus paracasei strain for suppressing periodontal disease inducing bacteria and use thereof}

It relates to a Lacticase bacillus paracasei strain having an activity to inhibit periodontal disease-causing bacteria and uses thereof.

Despite improvements in attitudes toward oral health, periodontal disease remains a prevalent problem. Current treatments for this disease include scaling and root planting, which seek to remove subgingival plaque and smooth tooth root surfaces to promote gum reattachment. Treatment is painful and often requires extensive home care adherence and follow-up visits. In addition, scaling and root planing are not prophylactic treatment options, making them less cost effective. Antibiotics are sometimes prescribed for treatment, but this can affect antibiotic resistance and the human microbiome.

Therefore, even according to the prior art, there is still a need for new periodontal disease prevention or treatment.

Life, Vol. 11, 268, pp.1-14 (Epub.2021.03.24.)

One aspect provides a Lacticaseibacillus paracasei subsp. tolerans LMT18-32 (Accession Number: KCTC14823BP) strain or a culture thereof having an activity to inhibit periodontal disease-causing bacteria.

Another aspect provides a composition for use in inhibiting periodontal disease-causing bacteria, including the strain or a culture thereof.

Another aspect provides a method for preventing or treating periodontal disease in a subject, comprising administering the strain or culture thereof to the subject.

One aspect provides a Lacticaseibacillus paracasei subsp. tolerans LMT18-32 (Accession Number: KCTC14823BP) strain or a culture thereof having an activity to inhibit periodontal disease-causing bacteria. The culture may be a culture medium of a microorganism or a combination thereof, a culture medium extract, or a protein extract. The extract may be a lysate obtained by lysing a microorganism or a combination thereof, or a supernatant remaining after centrifuging the same to remove a precipitate.

The periodontal disease causing bacteria may be Porphyromonas gingivalis . The strain may be viable in a solution containing an acid of pH 2 to 4 or 0.1% or more, eg, 1.5% or more, 2.0% or more, or 3.0% or more, such as bile acid. In addition, the strain may have active oxygen scavenging ability. The periodontal disease may be periodontitis or tooth decay. The acid may be an organic acid, an inorganic acid, or a combination thereof. The organic acid may be one constituting an object or generated during its metabolic process. For example, the organic acid may be amino acid, acetic acid, propionic acid, lactic acid, butyric acid, pentanoic acid, gluconic acid, or adipic acid.

Another aspect is a composition for use in inhibiting periodontal disease-causing bacteria, including Lactaseibacillus paracasei subsp. tolerans LMT18-32 (Accession Number: KCTC14823BP) strain or culture thereof. to provide.

In the composition, the culture may be a culture medium of a microorganism or a combination thereof, a culture medium extract, or a protein extract of a combination thereof. The extract may be a lysate obtained by lysing a microorganism or a combination thereof, or a supernatant remaining after centrifuging the same to remove a precipitate.

In the composition, the periodontal disease causing bacteria may be Porphyromonas gingivalis . The strain may be viable in a solution containing an acid of pH 2 to 4 or 0.1% or more, eg, 1.5% or more, 2.0% or more, or 3.0% or more, such as bile acid. In addition, the strain may have active oxygen scavenging ability. The periodontal disease may be periodontitis or tooth decay. The acid may be an organic acid, an inorganic acid, or a combination thereof. The organic acid may be one constituting an object or generated during its metabolic process. For example, the organic acid may be amino acid, acetic acid, propionic acid, lactic acid, butyric acid, pentanoic acid, gluconic acid, or adipic acid.

The composition may be a pharmaceutical composition for use in preventing or treating periodontal disease. The composition may include the strain as an active ingredient. The composition may be a food having an activity to prevent or improve periodontal disease.

The composition may include a carrier, diluent or excipient that is acceptable food or pharmaceutically.

Among the compositions, the microorganism or a combination thereof or a culture or extract thereof may be included in a “foodstically effective amount” or a “therapeutically effective amount”. In the above composition, "therapeutically effective amount" means an amount sufficient to exhibit a therapeutic effect when administered to a subject in need of treatment. The term “treatment” means treating a disease or medical condition, eg, a disease of obesity, in a subject, eg, a mammal, including a human, including: (a) preventing the disease or medical condition from occurring; ie, prophylactic treatment of the patient; (b) alleviating the disease or medical condition, ie, causing the disease or medical condition to be eliminated or reversed in a patient; (c) inhibiting the disease or medical condition, ie, slowing or stopping the progression of the disease or medical condition in a subject; or (d) alleviating a disease or medical condition in a subject. The "effective amount" can be appropriately selected by a person skilled in the art. The “effective amount” may be 0.01 mg to 10,000 mg, 0.1 mg to 1000 mg, 1 mg to 100 mg, 0.01 mg to 1000 mg, 0.01 mg to 100 mg, 0.01 mg to 10 mg, or 0.01 mg to 1 mg.

The composition may be administered orally. Accordingly, the composition may be formulated in various forms such as tablets, capsules, aqueous solutions or suspensions. In the case of tablets for oral use, excipients such as lactose and corn starch and lubricants such as magnesium stearate may be usually added. In the case of capsules for oral administration, lactose and/or dried cornstarch may be used as diluents. When oral aqueous suspensions are required, the active ingredient may be combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring agents may be added. The composition may be administered orally. The composition may be an oral dosage form.

Another aspect is Lacticase Bacillus paracasei subspecies tolerans ( Lacticaseibacillus paracasei subsp. tolerans ) LMT18-32 (accession number: KCTC14823BP) strain, periodontal disease of the subject, comprising administering a culture or extract thereof to the subject Provides a method for preventing or treating

The administration may be administration of the above composition. The strain is as described above.

In the above method, those skilled in the art can appropriately select an administration route according to the patient's condition. The administration may be oral or topical. The administration may be administered to the oral cavity, or to remain in the oral cavity after administration to the oral cavity.

In the above method, the dosage varies according to various factors such as the condition of the patient, the route of administration, the judgment of the attending physician, and the like, as described above. Effective doses can be estimated from dose-response curves obtained in vitro or in animal model tests. The ratio and concentration of the compound of the present invention present in the composition to be administered may be determined according to chemical characteristics, route of administration, therapeutic dose, and the like. The dosage may be administered to an individual in an effective amount of about 1 μg/kg to about 1 g/kg per day, or about 0.1 mg/kg to about 500 mg/kg per day. The dose may be changed according to the age, weight, sensitivity, or symptoms of the individual.

In the above method, the subject may be a mammal including a human. The subject may be, for example, a human, dog, cat, cow, pig, goat, or livestock such as a horse.

In the present specification, the strain or composition may be coated or encapsulated. The encapsulation may have the advantage of allowing controlled release, eg upon contact with water, or continuous re-release over a prolonged period of time. Furthermore, the composition may be protected from degradation to improve the shelf life of the product. Methods for encapsulation of active ingredients are well known in the art, and a number of encapsulating materials are available as well as methods of applying them to a composition according to specific needs.

In addition, the strain or composition may be in the form of a solution, suspension, emulsion, tablet, granule, powder or capsule.

The strain or composition may be toothpaste, tooth gel, tooth powder, tooth wash liquid, tooth wash foam, mouth wash, mouth spray, dental floss, chewing gum or lozenges.

A microorganism or a combination thereof or a culture or extract thereof according to one aspect may be used to prevent or treat periodontal disease.

A composition according to another aspect may be used to prevent or treat periodontal disease.

According to the method according to another aspect, periodontal disease can be effectively prevented or treated.

1 is a diagram showing the results of electrophoresis of the products obtained as a result of RAPD PCR using the genomes of the LMT18-32 strain and the KCTC3074 ^T strain as templates.
Figure 2 is a photograph showing the results of the growth inhibition test for Porphyromonas gingivalis KCTC5352 ^T of LMT18-32 cells and their culture.
Figure 3 is a diagram showing the results of analyzing the antioxidant activity of LMT18-32 and trolox by ABTS assay.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1. Isolation of strains

1. Isolation of Strains

Isolation of the strain took 100 g of kimchi, a traditional fermented food made at home, diluted in sterilized water, and homogenized for 5 minutes with a stomacher. The homogenized sample was diluted in stages and spread on MRS (Difco, USA) agar plate medium containing bromophenol blue (Sigma, USA) and cultured at 37 ° C for 2 to 3 days. The final strain was obtained by pure separation again. Hereinafter, this is referred to as LMT18-32. 16S rDNA phylogenetic analysis was performed as follows to confirm the phylogeny of the pure isolated lactic acid bacteria LMT18-32.

2. 16S rDNA analysis

The selected Lactobacillus LMT18-32 was subjected to PCR using primer sets of 27F (SEQ ID NO: 2) and 1492R (SEQ ID NO: 3) and the genome of LMT18-32 as a template to obtain 16S rDNA amplified products. The nucleotide sequence of the amplification product was confirmed through sequencing.

As a result, the 16S rDNA of LMT18-32 has the nucleotide sequence of SEQ ID NO: 1. In addition, the nucleotide sequence of the 16S rDNA was analyzed using NCBI blast (https://www.ncbi.nlm.nih.gov/). As a result of the phylogenetic tree analysis, LMT18-32 was the same as the Lactycasei bacillus paracasei subspecies tolerans. The 16S rDNA of LMT18-32 had 100.0% sequence identity with the Lacticase bacillus paracasei subspecies tolerance species. Therefore, the LMT18-32 strain was identified as a new strain belonging to the Lacticasei bacillus paracasei subspecies tolerance species.

3. Comparison of 16S DNA sequences of the selected LMT18-32 strain and type strain

In order to confirm the sequence identity of the selected Lactycasei bacillus paracasei subspecies tolerans LMT18-32 strain and the type strain, Lactycasei bacillus paracasei subspecies tolerans KCTC3074 ^T was commercially purchased from KCTC. As a result of comparing the sequences of the LMT18-32 strain and the KCTC3074 ^T strain, it was confirmed that the 16S rDNA sequence identity was 100.0%. In addition, in order to investigate the genetic relationship between the selected LMT18-32 strain and the type strain KCTC3074 ^T , each of the strains LMT18-32 and KCTC3074 ^T was subjected to OPA-13 (SEQ ID NO: 4) primers and the genome of each strain as a template. Thus, random amplification of polymorphic DNA (RAPD) PCR was performed to obtain an amplified product. The amplification products were subjected to electrophoresis on a 1.2% agarose gel containing RedSafe™ nucleic acid staining solution (iNtRON Biotechnology, Korea), followed by imaging with a ChemiDoc™ imaging system (Bio-Rad, USA) to obtain a DNA band pattern. 1 is a diagram showing the results of electrophoresis of the products obtained as a result of RAPD PCR using the genomes of the LMT18-32 strain and the KCTC3074 ^T strain as templates. As shown in Figure 1, the electrophoresis results between the two strains showed different band patterns.

The difference between the DNA band patterns of the Lactycasei bacillus paracasei subspecies tolerans LMT18-32 and the Lactycasei bacillus paracasei subspecies tolerans KCTC3074 ^T indicates that the two strains have a difference in genetic relatedness. Therefore, it was confirmed that the LMT18-32 strain is a new strain belonging to the subspecies tolerance species of Lacticase bacillus paracasei. Accordingly, the strain was named Lactaseibacillus paracasei subsp. tolerans LMT18-32, and it was submitted to the Korean Collection for Type Cultures (KCTC) at the Korea Research Institute of Bioscience and Biotechnology in December 2021. It was deposited on the 15th under the accession number KCTC14823BP.

Example 2. Measurement of antibacterial activity of lactic acid bacteria against periodontitis-inducing strain Porphyromonas gingivalis

In order to measure the antibacterial activity against periodontitis-inducing Porphyromonas gingivalis of the selected strain, Porphyromonas gingivalis KCTC5352 ^T was purchased and used from the Korea Cell Line Bank (Korean Collection for Type Cultures, KCTC) at the Korea Research Institute of Bioscience and Biotechnology. did First, Porphyromonas gingivalis was anaerobically cultured in TSA (Difco, USA) agar medium at 37 ° C. for 96 hours, and then Porphyromonas gingivalis was added at 1x10 ⁶ CFU / ml using Porphyromonas gingivalis TSA agar plates were prepared.

Next, in order to select lactic acid bacteria that inhibit the growth of Porphyromonas gingivalis, which causes periodontitis, the lactic acid bacteria isolated in Example 1 were cultured in MRS liquid medium at 37 ° C. for 24 hours. At this time, the OD600 was about 2.0. Then, each 10 μl of lactic acid bacteria from the cultured lactic acid bacteria was spotted on a TSA agar plate to which Porphyromonas gingivalis was added, and the lactic acid bacteria culture medium was centrifuged to recover only the culture medium, and only the supernatant was filtered with a 0.22 μm size filter. Pyromonas gingivalis was added to the TSA agar plate on top of the disk paper, and each 100 μl of the filtered culture medium was dispensed and anaerobically cultured at 37 ° C. for 96 hours. After culturing for 96 hours, the growth inhibition ring appeared by inhibiting the growth of Porphyromonas gingivalis was measured. The results are shown in Table 1 and FIG. 2.

Figure 2 is a photograph showing the results of the growth inhibition test for Porphyromonas gingivalis KCTC5352 ^T of LMT18-32 cells and their culture.

Lactobacillus Porphyromonas gingivalis KCTC5352 ^T growth inhibition ring (unit mm) Lacticase bacillus paracasei subspecies tolerans LMT18-32 cells 23 broth 32

In the case of using the Lactycasei bacillus paracasei subspecies tolerans LMT18-32 cell and culture medium, the length of the ring was 23mm and 32mm, respectively. This shows that 80 of 838 lactic acid bacteria used in the experiment showed excellent antibacterial activity against Porphyromonas gingivalis strain causing periodontitis, and among them, it has the best antibacterial activity.

Example 3. Investigation of morphological and fermentation characteristics of strains

1. Mycological characterization

Lacticase bacillus paracasei subspecies tolerance LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, was cultured on MRS plate medium, and colony morphologies were observed. The colony morphologies are shown in Table 2 below.

LMT18-32 shape circle size 0.7mm color cream color opacity opacity bump convex surface lubricity aerobic growth + anaerobic growth +

2. Sugar fermentation characteristics of selected lactic acid strains Sugar fermentation characteristics were investigated according to the supplier's experimental method using the API 50 CHL kit (Biomerieux, France). Table 3 shows the sugar fermentation characteristics of Lactasei bacillus paracasei subspecies tolerans LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains.

LMT18-32 glycerol - erythritol - D-arabinose - L-arabinose - D-ribose + D-Xylose - L-Xylose - D-Adonitol + Methyl-β D-xylopyranoside - D-galactose + D-glucose + D-fructose + D-mannose + L-Sorbose + L-rhamnose - dulcitol - inositol - mannitol + D-sorbitol + Methyl αα D-Mannopyranoside - Methyl α D-glucopyranoside - N-Acetylglucosamine + amygdalin - arbutin - esculin + Salicin + D-cellobiose + D-maltose + D-Lactose - D-Melibiose - D-saccharose + D-Trehalose + inulin + D-Melezitos + D-Raffinose - amidone - glycogen - xylitol - Gentiobios - D-Turanos + D-Likesauce + D-tagatose + D-Fucose - L-fucose - D-arabitol - L-arabitol - potassium gluconate + Potassium 2-ketogluconate - Potassium 5-ketogluconate -

As a result, it was confirmed that Lactase bacillus paracasei subspecies tolerans LMT18-32 uses a total of 22 sugars.

Example 4. Stability

1. Investigation of acid resistance of lactic acid bacteria

In order for the lactic acid bacteria LMT18-32 to exhibit efficacy as probiotics in the oral cavity, they must pass through or be maintained in a low pH environment after ingestion. For example, the strain may be maintained or grown in an acidic environment excreted by bacteria living in plaque, which is a bacterial film on the tooth surface, such as S. mutans . In order to investigate the acid resistance of Lacticase bacillus paracasei subspecies tolerans LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, it was inoculated into MRS liquid medium and cultured at 37° C. for 24 hours. Next, the pH 2.5 MRS liquid medium adjusted to pH 2.5 using HCl (Sigma, USA) was prepared, and the cultured lactic acid bacteria were inoculated into the pH 2.5 MRS liquid medium and cultured at 37 ° C. for 2 hours. At this time, the lactic acid bacteria immediately after inoculation were diluted and plated on the MRS plate medium, and the lactic acid bacteria cultured for 2 hours in the pH 2.5 MRS liquid medium were also plated on the MRS plate medium and cultured at 37 ° C. for 24 hours. After 24 hours, the results of counting the number of lactic acid bacteria colonies on the MRS plate medium are shown in Table 4.

LMT18-32 (CFU/mL) MRS (pH 6.8) 4.0×10 ⁹ MRS (pH 2.5) 3.9×10 ⁹

As a result, Lacticase bacillus paracasei subspecies tolerans LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, showed a high survival rate by maintaining a bacterial count of 3.9×10 ⁹ CFU/ml. The characteristics of these lactic acid bacteria can be expected to be maintained or grown, that is, to have acid resistance even in an acidic pH environment in which tooth decay is induced. It is known that acid is released by bacteria such as S. mutans in the plaque where tooth decay occurs, resulting in acidity. In addition, since the number of lactic acid bacteria was maintained at a pH lower than pH 3, which is close to the physiological pH of the stomach, it is possible to stably maintain the number of viable bacteria even at a low pH due to gastric acid secretion, and it can be expected that the intestinal reach rate is high when ingested.

2. Investigation of bile resistance of lactic acid bacteria

In order to investigate the bile of lactic acid bacteria, the following experiment was conducted. After inoculation of Lacticase bacillus paracasei subspecies tolerans LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, in MRS liquid medium, it was incubated at 37° C. for 24 hours. Next, considering that the concentration of bile salts in the intestinal tract is around 0.1%, an MRS liquid medium containing 0.3% bile salt (Sigma, USA) was prepared, and the cultured lactic acid bacteria were inoculated into the 0.3% Bile salt MRS liquid medium. and incubated for 2 hours at 37°C.

At this time, the lactic acid bacteria immediately after inoculation were diluted and plated on the MRS plate medium, and the lactic acid bacteria cultured for 2 hours in 0.3% Bile salt MRS liquid medium were also plated on the MRS plate medium and cultured at 37 ° C. for 24 hours. After 24 hours, the results of counting the number of lactic acid bacteria colonies on the MRS plate medium are shown in Table 5.

LMT18-32 (CFU/mL) MRS (0% bile salt) 4.0×10 ⁹ MRS (0.3% bile salt) 1.4×10 ⁹

As a result, Lacticase bacillus paracasei subspecies tolerance LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, showed a high survival rate by maintaining a bacterial count of 1.4×10 ⁹ CFU/ml. Therefore, considering that the concentration of bile salts in the intestine is around 0.1%, it can be expected that the survival rate in the intestine is sufficiently high.

Example 5. ABTS radical scavenging activity of lactic acid bacteria

In periodontal disease, reactive oxygen species (ROS) are generated as infectious and inflammatory processes occur continuously, and these reactive oxygen species exacerbate periodontitis (J Indian Soc Periodontol. 2013 Jul-Aug; 17(4): 411- 416.). At this time, antioxidants help to improve periodontal disease by removing active oxygen and reducing oxidative stress.

Therefore, in order to confirm the antioxidant efficacy of Lacticase bacillus paracasei subspecies tolerans LMT18-32, which has excellent antibacterial activity against periodontitis-inducing strains, ABTS (2,2′ -Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) An assay was performed.

In this analysis, ABTS loses electrons by potassium persulfate to show a deep bluish green color, but when antioxidants are present, the color becomes pale due to its electron donating ability. This process is a method for measuring antioxidant activity. First, 14mM ABTS (38.4mg/10ml; Sigma, USA) and 6mM potassium (8.1096mg/10ml; Sigma, USA) were mixed in a 1:1 ratio and reacted in the dark at room temperature for 16 hours to form positive ions (ABTS ⁺ ). . The ABTS+ solution thus prepared was diluted 1/10 and used. After culturing the lactic acid bacteria at an OD600 of about 2.0, centrifugation was performed to recover only the culture medium, and only the supernatant was filtered with a 0.22 μm size filter. 20 μl of the filtered culture medium and 180 μl of the ABTS ⁺ solution were reacted in the dark in a 96-well plate for 10 minutes. Thereafter, the sample after the reaction was measured for absorbance at 734 nm using a microplate reader, and trolox (1 mg/ml; Sigma, USA), a precursor of vitamin E, an antioxidant, was used as a positive control. Figure 3 is a diagram showing the results of analyzing the antioxidant activity of LMT18-32 and trolox by ABTS assay.

As shown in Figure 3, the 10% concentration of the culture medium of LMT18-32 showed a high antioxidant activity of 82.9%. Therefore, the LMT18-32 acts as an active oxygen inhibitor and can help improve oral health such as preventing or treating periodontal disease.

SEQUENCE LISTING <110> Medytox Inc. <120> Lacticaseibacillus paracasei strain for suppressing periodontal disease inducing bacteria and their use <130> PN142655KR <160> 4 <170> PatentIn version 3.2 <210> 1 <211> 1403 <212> DNA <213> Lacticaseibacillus paracasei subsp. tolerans LMT18-32 <400> 1 gagttctcgt tgatgatcgg tgcttgcacc gagattcaac atggaacgag tggcggacgg 60 gtgagtaaca cgtgggtaac ctgcccttaa gtgggggata acatttggaa acagatgcta 120 ataccgcata gatccaagaa ccgcatggtt cttggctgaa agatggcgta agctatcgct 180 tttggatgga cccgcggcgt attagctagt tggtgaggta atggctcacc aaggcgatga 240 tacgtagccg aactgagagg ttgatcggcc acattgggac tgagacacgg cccaaactcc 300 tacgggaggc agcagtaggg aatcttccac aatggacgca agtctgatgg agcaacgccg 360 cgtgagtgaa gaaggctttc gggtcgtaaa actctgttgt tggagaagaa tggtcggcag 420 agtaactgtt gtcggcgtga cggtatccaa ccagaaagcc acggctaact acgtgccagc 480 agccgcggta atacgtaggt ggcaagcgtt atccggattt attgggcgta aagcgagcgc 540 aggcggtttt ttaagtctga tgtgaaagcc ctcggcttaa ccgaggaagc gcatcggaaa 600 ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt gaaatgcgta 660 gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc 720 tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg taaacgatga 780 atgctaggtg ttggagggtt tccgcccttc agtgccgcag ctaacgcatt aagcattccg 840 cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc cgcacaagcg 900 gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggtct tgacatcttt 960 tgatcacctg agagatcagg tttccccttc gggggcaaaa tgacaggtgg tgcatggttg 1020 tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttatgac 1080 tagttgccag catttagttg ggcactctag taagactgcc ggtgacaaac cggaggaagg 1140 tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt gctacaatgg 1200 atggtacaac gagttgcgag accgcgaggt caagctaatc tcttaaagcc attctcagtt 1260 cggactgtag gctgcaactc gcctacacga agtcggaatc gctagtaatc gcggatcagc 1320 acgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgagagttt 1380 gtaacacccg aagccggtgg cgt 1403 <210> 2 <211> 20 <212> DNA <213> artificial <220> <223> primer <400> 2 agagtttgat cmtggctcag 20 <210> 3 <211> 19 <212> DNA <213> artificial <220> <223> primer <400> 3 ggttaccttg ttacgactt 19 <210> 4 <211> 10 <212> DNA <213> artificial <220> <223> primer <400> 4 cagcacccac 10

Claims (7)

Lacticaseibacillus paracasei subsp. tolerans LMT18-32 (accession number: KCTC14823BP) strain having an activity to inhibit periodontal disease-causing bacteria, wherein the periodontal disease-causing bacteria is Porphyromonas gingibal Less ( Porphyromonas gingivalis ) The strain. A culture of Lactaseibacillus paracasei subsp. tolerans LMT18-32 (Accession Number: KCTC14823BP) strain, which has activity to inhibit periodontal disease-causing bacteria, wherein the culture is a microorganism, culture medium, It is a microbial lysate or a supernatant of the microbial lysate, and the periodontal disease-causing bacteria is Porphyromonas gingivalis ( Porphyromonas gingivalis ). A composition for use in inhibiting periodontal disease-causing bacteria, comprising Lactaseibacillus paracasei subsp. tolerans LMT18-32 (Accession Number: KCTC14823BP) strain or culture thereof, The disease-causing bacteria are Porphyromonas gingivalis ( Porphyromonas gingivalis ), and the culture is a microorganism, a culture medium, a microbial lysate or a supernatant of a microbial lysate, and a pharmaceutical composition for use in preventing or treating periodontal disease. delete The method according to claim 3, wherein the periodontal disease is a composition of periodontitis or caries. delete A composition for use in inhibiting periodontal disease-causing bacteria, including Lactaseibacillus paracasei subsp. tolerans LMT18-32 (accession number: KCTC14823BP) strain or culture thereof, wherein the culture The water is a microorganism, a culture medium, a microbial lysate or a supernatant of a microbial lysate, and the periodontal disease-causing bacteria is Porphyromonas gingivalis , and a composition that is a food having an activity to prevent or improve periodontal disease .

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