KR20130002533A - Lactic acid bacteria group hindering growth of helicobactor pyloli - Google Patents

Abstract

PURPOSE: Lactic acid bacteria which are derived from kimchi and suppress helicobacter pylori are provided to prevent and treat gastritis, gastric ulcer, and duodenal ulcer. CONSTITUTION: A Leuconostoc mesenteroides LAB kw5(KACC91648P) suppresses growth of helicobacter pylori. A composition for suppressing helicobacter pylori contains the strain or a culture thereof as an active ingredient. The composition additionally contains a Streptococcus thermophilus LAB kw15(KACC91649P) or a culture thereof as an active ingredient. Probiotics contain Leuconostoc mesenteroides LAB kw5(KACC91648P) or the culture and Streptococcus thermophilus LAB kw15(KACC91649P) or the culture as active ingredients. A food additive contains Leuconostoc mesenteroides LAB kw5(KACC91648P) or the culture.

Description

Lactic acid bacteria inhibiting the growth of Helicobacter pylori {LACTIC ACID BACTERIA GROUP HINDERING GROWTH OF HELICOBACTOR PYLOLI}

The present invention relates to kimchi-derived lactic acid bacteria that inhibit the growth of Helicobacter pylori.

Helicibacter pylori is closely related to chronic gastroduodenal disease, and is known to be a causative agent of chronic gastritis, gastric ulcer, peptic ulcer, and gastric cancer by inhabiting the junction between gastric mucosal epithelial cells [Parsonnet J, Friedman GD, Vandersteen DP , chang Y, Vogelman JH, Drentreich N, Sibley RK. 1991.Helicibacter pyroli infection and the risk of gastric carcinoma. N Engl J Med 325: 1127-1131 .; Lighting agent, Lee Kwang-ho. 1996 Helicibacter pyroli The cause of gastric cancer caused by infection. Medical Postgraduates. 3 (24): 16-157 .; Cover. TL and MJ Blaser. 1995. Helicibacter pyroli : Abacterial cause of gastritis, pepticulcer disease, and gastric cancer. ASMMews 61: 21-26; Lee, A., J. Fox, and S. Hazell. 1993. Pathogenicity of Helicibacter pyroli : Apersprctive. Infect . Immun. 61: 1601-1610 .; Marshall, BJ 1994. Helicibacter pyroli . Am . J. Gastroenterol . 89: S116-S128 .; World Health Organization. 1994. IARC Monographs on the Evaluation of Carcinogenic Risks to Human. 61: 177-240. Characteristic characteristics of Helicobacter pylori form a colony of gastric epithelial cells, and by using strong urease activity to decompose plasma effluent or urea in gastric mucosa into ammonia and bicarbonate to alkalinize around the strain, Can live in acidic conditions [Mobely HL, Island MD, Hansinger RP. 1995. Molecular biology of mirobial. J Microbiol Rev 59: 169-182. In addition, unipolar flagella allow the Helicobacter pylori to move quickly into the viscous layer, thus avoiding prolonged exposure in the acidic stomach. However, the Helicobacter pylori can be disinfected at about 80% by the administration of three or more drugs, but the problem is that the emergence of resistant bacteria due to the increased use of antibiotics, and the Helicobacter pylori bactericidal does not significantly reduce the incidence of gastric cancer. Inflammation persists. Recently, there is a growing interest in developing foods, extracts, vaccines, etc., in addition to drug elimination treatment, because of the lack of clear guidelines for which patients should be disinfected. Increasingly, the use of foods and probiotics can be used to treat them [Kim, NY 2006. The effect of antibiotic resistance on the eradication of Helicibacterpyroli . Kor , J, Gastroenterol . 47: 82-86.]. In particular, the genus Lactobacillus spp.) has been reported to be effective in inhibiting Helicobacter pylori when a culture of lactic acid bacteria belonging to human or laboratory animals is used [Serin, AL 2001. Antagonistic activity against Helicobacter pylori infection by the lactic acid bacteria. J. Kor . Public Health . assoc . 27: 5-12. Lactobacillus spp.) is known to produce significant lactic acid during metabolism. Since Helicobacter pylori live in the stomach with low hydrogen ion concentration, the lactic acid pKa has low lactic acid, which may be present in the non-liquid state, which may act as an inhibitory effect on Helicobacter pylori. However, lactic acid bacteria secrete antimicrobial substances and may be more effective in inhibiting the growth of Helicobacter pylori by these substances.

Currently, fermented milk using lactic acid bacteria having a helicobacter pylori inhibitory ability has been developed and sold in Korea, but lactic acid bacteriocin has not been studied yet.

Bacteriocin is a protein or peptide-based antibacterial substance produced by various bacteria and is known to be secreted to inhibit the growth of microorganisms different from or similar to bacteriocin producing bacteria in a specific growing environment [Klaenhammer TR.1988. Bacteriocin of lactic acid bacteria Biochimie 70: 337-349.]. In addition, bacteriocin is harmless to humans, non-residual, biosynthetic directly from plasmids or chromosomes (choromosome) has the advantage of easy biotechnological applications such as genetic engineering. In particular, many lactic acid bacteria that are widely used in food fermentation produce a variety of bacteriocins. Of these, nisin is used commercially and is best known [Oh SJ, Lee JH, Kim GT, Shin JG, baek YJ. 2003.Anti-carcinogenic activity of a bacteriocin produced by Lactococcus sp. HY449. Food Sci Biotechnol 12: 9-12.]. Since lactic acid bacteria or bacteriocins produced by lactic acid bacteria inhibit several gram-positive bacteria, the possibility of developing them as food preservatives replacing chemical preservatives is very high [Choi YO, Ahn C. 1997. Plasmid-associated bacteriocin production y Leuconostoc sp. LAB145-3 Aisolated from Kimchi. J Ind Microbiol 2: 319-322.].

The present invention isolates and identifies lactic acid bacteria having excellent Helicobacter pylori inhibitory ability from kimchi and decay, and confirms the Helicobacter pylori growth inhibitory substance secreted by the isolated lactic acid bacteria to be used for the prevention and treatment of gastrointestinal diseases related to Helicobacter pylori. It was intended to be used as a material.

Accordingly, the present invention is a Leuconostoc that inhibits the growth of Helicobacter pylori ( Leuconostoc) mesenteroides ) LAB kw5 strain [KACC91648P] to provide.

In addition, the present invention has another object to provide a composition for inhibiting Helicobacter pylori as an active ingredient of the strain or its culture.

In addition, the present invention is another object to provide a medicament for the preparation, probiotics, food additives or gastritis, gastric ulcer or duodenal ulcer prevention and treatment comprising the strain or its culture.

In order to achieve the above object, the present invention is Helicobacter pylori ( Helicobacter Leuconostoc mesenteroides LAB kw5 strain [KACC91648P], which inhibits the growth of pylori ).

In addition, the present invention provides a composition for inhibiting Helicobacter pylori as an active ingredient of the strain or its culture.

In addition, the present invention provides a medicament for the preparation and prophylactic agent, probiotic, food additive or gastritis, gastric ulcer or duodenal ulcer prevention and treatment comprising the strain or its culture.

Leuconostoc mesenteroides LAB kw5 strain of the novel lactic acid bacterium of the present invention has the characteristics of inhibiting the growth of Helicobacter pylori and excellent antibacterial activity. Therefore, the lactic acid bacteria of the present invention can be used in various forms for the preparation of formal preparations, probiotics, antibacterial agents, feed additives, food additives. In particular, it is effective in the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori.

1 (A) is Leuconostoc mesenteroides ) confirmed the growth inhibition (halo) of the Helicobacter pylori by the LAB kw5 strain, (B) Streptococcus thermophilus ) was confirmed to inhibit the growth of Helicobacter pylori by LAB kw15 strain.
Figure 2 shows the results of 16s rRNA gene amplification of the Leuconostoc mesenteroides LAB kw5 strain and Streptococcus thermophilus LAB kw15 strain isolated from kimchi [Lane 1: L. mesenteroides KCTC 3105, Lane 2: L. mesenteroides LAB kw5, Lane 3: S. thermophilus LAB kw15, M: 100 bp DNA ladder].
3 is different from L. mesenteroides LAB based on 16s rRNA sequence The phylogenetic relationship with LAB is shown.
4 L. mesenteroides LAB kw5 and S. thermophilus After adjusting the culture supernatant of LAB kw15 to pH 7.0, H. pylori The effect of H. pylori on the growth of H. pylori with time was added to the culture broth [(A) H. pylori KCTC12083, (B) H. pylori 52] .
5 L. mesenteroides LAB kw5 and S. thermophilus LAB kw15 was incubated with H. pylori to confirm the change in growth inhibition according to incubation time [Lac5: L. mesenteroides LAB kw5; Lac 15: S. thermophilus LAB kw15; kctc: H. pylori KCTC12083, L5: L. mesenteroides LAB kw5; L15: S. thermophilus LAB kw15; 52: H. pylori 52].
6 is L. mesenteroides of the present invention H. pylori of LAB kw5 strain and known strain L. gasseri 250 It showed antimicrobial activity [(A) H. pylori KCTC12083 + L. gasseri 250; (B) H. pylori KCTC12083 + L. mesenteroides LAB kw5].

Lukonostok mesentroid LAB kw5 of the present invention is characterized by inhibiting the growth of Helicobacter pylori and having antibacterial activity. In particular, Lukonostok mesentroid LAB kw5 of the present invention isolated from kimchi has the activity of inhibiting the growth of Helicobacter pylori, it has the characteristics of having an antibacterial activity better than L. gasseri 250.

Suppressing the growth of the Helicobacter pylori as described above, and has all of the features of excellent antibacterial activity Lukonostok mesoidoid LAB kw5 strain is provided for the first time in the present invention.

Therefore, the lactic acid bacteria of the present invention can be used as a composition for inhibiting Helicobacter pylori, the use for improving the health of humans and animals, that is, suits, probiotics, food additives.

The composition may include a crushed cell wall fraction, live bacteria, dead bacteria, dry bacteria or cultures of the luconosestock methoidoid of the present invention as an active ingredient, and may further include an excipient or a carrier. The culture includes a culture solution itself cultured in a liquid medium, a filtrate (centrifuged supernatant) from which the strain is removed by filtration or centrifugation of the culture solution. The content of luconosestock mecentoid LAB kw5 in the composition may vary depending on the use and formulation of the composition. Formal, antimicrobial or probiotic compositions according to the invention may be prepared and administered in a variety of formulations and methods. For example, lukonostume methroid LAB kw5 or its culture may be mixed with carriers and flavorings commonly used in the pharmaceutical arts for tablets, troches, capsules, elixirs. ), Syrup, powder, suspension or granule. As the carrier, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent and the like can be used. The mode of administration may be oral, parenteral or application, but preferably oral administration. In addition, the dosage may be appropriately selected depending on the absorbency, inactivation rate and excretion rate of the active ingredient in the body, age, sex, condition, etc. of the recipient.

In addition, the luconostoke mesentroid or the culture thereof according to the present invention can be used as an agent for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer.

The bacterium Helicobacter pylori ( H. pylori ) was first reported by Warren and Marshall in 1982 as a causative agent of chronic gastritis, gastric ulcer, duodenal ulcer, gastritis and peptic ulcer, and antibiotics, natural products and lactic acid bacteria. Inhibiting the growth of Helicobacter pylori has been known to prevent and treat the diseases caused by this [Midole et al., J. of Applied bacteriology, 79, 475-479, 1995; Bhatia et al., J. of Clinical Microbiology, Oct., 2328-2330, 1989].

In the present invention, it was confirmed through the growth inhibitory ring experiment that the ruconostock mesentroid of the present invention has an activity of inhibiting the growth of Helicobacter pylori (see FIG. 1). Therefore, the leukostock memetroid or the culture thereof of the present invention having the activity of inhibiting the growth of Helicobacter pylori can be used as an agent for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer due to infection of Helicobacter pylori.

Lukonostok mesentroid or its culture according to the present invention can be used as a food additive for food such as kimchi, beverages, baby food, dairy products.

Hereinafter, the present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the following examples.

Reference Example :. Strains and Mediums Used

The strain used in this experiment was Helicobacter pylori KCTC 12083 was distributed at BRC and wild type H. pylori 52 was sold by Gyeongsang National University College of Medicine.

L. mesenteroides And lactic acid bacteria in MRS broth or culture is a Lactobacilli MRS agar (Difco Laboratory, Detroit, MI, US) 37 ℃, 48 to 72 hours were incubated at, H. pylori is made 10% horse serum was added to 0.05% L- cysteine After inoculation to the added Brucella broth or Brucella agar (Difco Laboratory, Detroit, MI, US) 48 hours in a microaerobic chamber (Ruskin Technologies, Jouan, France) or 10% CO ₂ incubator (Sanyo, Osaka, Japan) at 37 ℃ Incubated.

Example  1: Separation and Identification

1) Lactobacillus Isolation

In order to separate lactic acid bacteria from kimchi, 30 kinds of kimchi, which are manufactured and sold directly in traditional markets and department stores, were collected, and five kinds of fallen foods, which are unique traditional foods in Andong, were collected.

Depravity is a fermented beverage of lactic acid in some parts of Korea that uses rice wine spawn in milk. 25 g of these samples were mixed in 225 mL of 0.1% peptone water, homogenized using a stomacher, and then diluted to 10. The cells were incubated at 37 ° C. for 48 to 72 hours after smearing in MRS agar. And morphologically different among the cultured colonies were selected and plated in MRS agar and cultured by pure separation. Thereafter, the susceptible strains of lactic acid bacteria were examined for morphological characteristics through a microscope, Gram test and catalase test, and biochemical characteristics such as glycosylation were analyzed using API kit 50CHL (Biomerieux Co., France).

2) 16s rDNA  Identification through sequencing

The isolated lactic acid bacteria were subjected to 16s rRNA sequence analysis for glycosylation characteristics and primary identification after final identification using API kit 50CHL. Lactic acid bacteria isolated for sequencing were incubated in Lactobacilli MRS broth, 1.5 mL of the culture solution was centrifuged, and washed with 0.8% sterile saline. The chromosomal DNA was extracted using genomic DNA kit and used as template DNA for PCR. F-341 (CCTACGGGAGGCAGCAG) and 786-R (GACTACCAGGGTATCTAATC), universal primers for bacterial 16s rRNA, were used. After PCR, the PCR product was identified by 450 bp amplification by electrophoresis and purified using a QIAquick PCR purification kit (QIAGEN, Hilden, Germany). The base sequence was performed by a Dye Terminator Cycle Sequencing Ready Reaction Kit (Biosystems, CA, USA) and an ABI 3700 sequencer (Biosystems, CA, USA), and primers were used in the same manner as above. Homology analysis of 16S rRNA sequences was compared to DNA databases using the BLAST search program of the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/). Genetic analysis was performed using Clustal X, BioEdit, and MEGA 4 (www.megasoftware.net/) to confirm the genetic distance between the sequences and the phylogenic tree.

3) Results

The lactic acid bacteria were isolated from five samples of fermented milk, and 30 samples of kimchi purchased from the market. The isolates were isolated according to the morphological characteristics of colonies cultured in Lactobacilli MRS agar, and the bacteria suspected of lactic acid bacteria were selected through gram test, catalase test, and morphological characteristics through microscopic observation. The biochemical characteristics such as sugar fermentation characteristics were confirmed by using API 50 CHL kit, and the results were confirmed through the identification program, and 15 strains of strains having more than 90% identification result were selected.

This strain is Lactobacillus curvatus 5 weeks , Leuconotoc mesenteroides 2 weeks, Lactococcus lactis 2 weeks, Weissella confusa 3 weeks, Lactobacillus fermentum 1 week, Streptcoccus thermophilus 1 week, Lactobacillus plantarum One week was isolated and identified. The culture medium of this isolate was adjusted to neutral and diluted, and the diluted culture solution was plated on agar medium mowed with the standard strain and H. pyroli , the isolate strain. At this time, 5, 9 isolates of halo-forming strains that inhibit H. pyroli were isolated from each sample. Spot on the lawn method, disk diffusion, well diffusion method was performed to confirm the clear halo (halo not shown). Finally, the strain that forms the largest inhibitory ring among the inhibitory ring-forming strains was selected by one strain from each sample.

The growth supernatant of H. pylori was confirmed by adjusting the culture supernatant of lactic acid bacteria to pH 7.0 in order to confirm the inhibitory effect of H. pylori on the growth of lactic acid bacteria cultures. Otherwise the resulting culture supernatant was confirmed to inhibit the growth of H. pylori due to the lactic acid bacteria isolated from Kimchi, the Fall and when adjusted to pH 7.0 was able to determine both the growth inhibition ring of H. pylori [1]. H. pylori according to lactic acid bacteria Inhibitory activity was found to be different, especially H. pylori of Leuconostoc mesenteroides LAB kw5. The greatest inhibition was observed. From these results, the inhibition of growth of H. pylori is known to be a complex action of low pH caused by lactic acid bacteria and lactic acid [Serin, AL 2001. Antagonistic activity against Helicobacter pylori infection by the lactic acid bacteria. J. Kor . Public Health . assoc . 27: 5-12 .; Lee. HS, SH Park, HK Chung, and TB Choe. 1993.Antitumor effect of cell wall component purified from Lactobacillus plantarum KK. J. Gen. Eng . 5: 22-28.]. In this experiment, the antimicrobial activity of H. pylori was observed even after adjusting to pH 7.0. Jung et al. [Jung HK, Kim ER, Juhn SL. 2001.Selection of Lactic Acid Bacteria Specifically Inhibiting the Growth of Helicibacter pyroli. 37: 152-153.] is the H. pylori It is reported that the most resistant strain against the strain did not produce a large amount of lactic acid, and the isolated lactic acid bacteria produced antimicrobial substances that inhibit the growth of H. pylori .

The glycosylation of the final isolate strain [Table 1] and 16s rRNA sequencing analysis showed that the LAB kw5 isolated from Kimchi was Leuconostoc. mesenteroides [SEQ ID NO: 1], LAB kw15 isolated from fallen Streptococcus thermophilus [SEQ ID NO: 2] was identified [Fig. 2, 3].

The isolated Leuconostoc mesenteroides LAB kw5 strain and Streptococcus thermophilus LAB kw15 were assigned accession numbers as KACC91648P and KACC91649P as of April 5, 2011, National Institute of Agricultural Science.

Example  2: isolated lactic acid bacteria H. pylori Growth inhibition against

H. pylori , the standard strain KCTC12083 and wild type H. pylori 52 were incubated for 48 hours in Brucella agar each with horse serum, and then suspended in 1 ml Brucella broth with horse serum in 3McFaland units and plated. The lactic acid bacteria cultured in MRS broth were centrifuged (10,000 × g, 5 min), the supernatant was adjusted to pH 7.0 using 1N NaOH, and then sterilized using a 0.22 μm Syringe filter (Millipore, Billerica, MA). 10 μl each of the bacteriostatic solution was added to the medium in which H. pylori was cultured, followed by spot on the lawn method, disk diffusion, and well diffusion method, respectively.

The culture supernatants of the isolated L. mesenteroides LAB kw5 and S. thermophilus LAB kw15 were adjusted to pH 7.0 and added to the H. pylori culture to determine the effect on the growth of H. pylori over time [Fig. 4]. If the addition of culture the culture supernatant in the H. pylori strains as compared to the time the culture supernatant did not added with the wild-type H. pyroli In all, it was confirmed to inhibit the growth. L. mesenteroides LAB kw5 H. pylori More than 30% of H. pylori compared to KCTC12083 alone 52 showed more than 25% higher growth inhibitory activity compared to the cultured section, and S. thermophilus LAB kw15 was found to be H. pylori KCTC12083 is about 30% higher than the cultured section, H. pylori Compared to the 520,000 cultured section, more than about 20% of the growth inhibitory activity was shown. Growth inhibition was greater than wild-type function in a standard strain of H. pylori, than S. thermophilus L. mesenteroides kw15 LAB LAB kw5 higher H. pylori Activity inhibitory ability was confirmed. However, the difference between L. mesenteroides LAB kw5 and S. thermophilus LAB kw15 production and secretion ability is not significant.

Example  3: term of lactic acid bacteria culture medium H. pylori Effect and mixed culture

H. pylori prepared by the same method as above to determine the effect of lactic acid bacteria culture medium having antibacterial effect on H. pylori on the growth of H. pylori The same amount of the culture suspension and lactic acid bacteria culture supernatant whose pH was adjusted to 7.0 was added, and the growth of H. pylori over time was compared using a microplate reader. The supernatant of H. pylori and lactic acid bacteria was added to a tissue culture test plate (SPL LIFE SCIENCES, Namyangju, Korea) and the absorbance of the culture solution was measured at 660 nm at 3 hours intervals for 3 hours at 37 ° C in a CO ₂ incubator. . The mixed culture of lactic acid bacteria and H. pylori was suspended in a single colony of H. pylori in 50 ml Brucella liquid medium, and cultured for 36 hours at 37 ° C. in a CO ₂ incubator. After that H. pylori is cultured and put as the lactic acid is diluted at a constant ratio to the culture broth measured the number of H. pylori and the lactic acid bacteria with time and H. pylori The inhibition was examined.

The growth inhibition of the lactic acid bacteria and H. pyroli were cultured and measured according to the culture time. However, when the growth time is different because the growth time differs from Lactobacillus to 24 to 36 hours, and H. pylori to 48 to 72 hours, when H. pylori has a liquid culture of 36 hours, inoculate the lactic acid bacteria at 10 ³ CFU / ml and mix culture. Was done. Lactic acid bacteria increased rapidly after 6 hours of incubation and reached its peak at 24 hours. 18 hours ago LAB kw15, 18 hours after the culture is more fast speed of L. mesenteroides LAB kw5. The slight difference in H. pylori, but lactic acid was retained, indicating the inhibition of the maximum force of about 6-12 hours in the case of beginning to secrete inhibitors from 0 to 6 hours after the culture, particularly, L. mesenteroides LAB kw5 . However, S. thermophilus LAB kw15 was found to weaken the inhibition by about 20-30% after 12 hours. This is thought to slow the growth rate rather than inhibit H. pylori . Interestingly, these isolated lactic acid bacteria were similar in their ability to produce antimicrobial substances, but showed very different forms of H. pyroli inhibition in mixed cultures. L. mesenteroides It is thought that LAB kw5 strain has a high inhibitory activity against H. pyroli growth in addition to the antibacterial substances secreted by this strain, producing lactic acid or other H. pyroli inhibitors [Serin, AL 2001. Antagonistic activity against Helicobacterpylori infection by the lactic acid bacteria. J. Kor . Public Health . assoc . 27: 5-12 .; Lee. HS, SH Park, HK Chung, and TB Choe. 1993.Antitumor effect of cell wall component purified from Lactobacillus plantarum KK. J. Gen. Eng . 5: 22-28.]. Therefore, L. mesenteroides LAB kw5 appears to be a lactic acid bacterium with good function in controlling H. pyroli .

Example  4:  L. mesenteroides LAB kw5 Wow L. gasseri  250 H. pylori Antimicrobial Activity Results

1) Cultivation of Strains Used

The strain used in this experiment was Helicobacter pylori KCTC 12083 was distributed at a BRC. L. mesenteroides and L. gasseri LAB kw5 cultured in 250 of 0.05% was added to L- cysteine cultured made Lactobacilli MRS broth or MRS agar 37 ℃, 48 ~ 72 hours at (Difco Laboratory, Detroit, MI, US), H . pylori is 10% horse serum is added to a Brucella broth or Brucella agar at 37 ℃ after inoculation in (Difco Laboratory, Detroit, MI, US) microaerobic chamber (Ruskin Technologies, Jouan, France) and 10% CO ₂ constant temperature incubator (Sanyo , Osaka, Japan) was incubated for 48 hours.

2) H. pylori  Antimicrobial effect against

H. pylori KCTC12083, a standard strain, was incubated for 48 hours in Brucella agar with horse serum, and then suspended in 1 ml Brucella broth with horse serum in 3McFaland units. The L. gasseri 250 and L. mesenteroides LAB kw5 incubated in MRS broth were centrifuged (10,000 × g, 5 min) and the supernatant was adjusted to pH 7.0 using 1N NaOH and 0.22 μm Syringe filter (Millipore, Billerica, Sterilization using MA). 10 μl each of the bacteriostatic solution was added to the medium in which H. pylori was cultured, followed by spot on the lawn method, disk diffusion, and well diffusion method, respectively.

As a result, L. mesenteroides LAB kw5 showed more excellent antimicrobial activity compared to L. gasseri 250, which is well known as lactic acid bacteria having an antimicrobial effect against H. pylori [Fig. 6].

Therefore, L. mesenteroides LAB kw5 appears to be a lactic acid bacterium with good control of H. pylori .

division L. mesenteroides
LAB kw5 L. gasseri 250 H. pylori KCTC
12083 disk 3 mm 0 mm well 18 mm 15 mm spot 18 mm 14 mm

National Institute of Agricultural Science, Genetic Resources Center, Korea KACC91648P 20110405 National Institute of Agricultural Science, Genetic Resources Center, Korea KACC91649P 20110405

<110> BKBIO.CO., Ltd.          Kyungwon University-Industry Collaboration Foundation <120> LACTIC ACID BACTERIA GROUP HINDERING GROWTH OF HELICOBACTOR          PYLOLI <130> a <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 844 <212> DNA <213> Leuconostoc mesenteroides LAB kw5 <400> 1 nngaaggctt tcgggtcgta aagcactgtt gtatgggaag aacagctaga ataggaaatg 60 attttagttt gacggtacca taccagaaag ggacggctaa atacgtgcca gcagccgcgg 120 taatacgtat gtcccgagcg ttatccggat ttattgggcg taaagcgagc gcagacggtt 180 tattaagtct gatgtgaaag cccggagctc aactccggaa tggcattgga aactggttaa 240 cttgagtgca gtagaggtaa gtggaactca tgtgtagcgg tggaatgcgt agatatatgg 300 aagaacacca gtggcgaagg cggcttactg gactgcaact gacgttgagg ctcgaaagtg 360 tgggtagcaa acaggattag accgggggga tatctccatc aaacttgagc aaaagcttat 420 ggaaccatcc gcgagggaaa aagccttgga ctgaagcatg tgttgcaaga aatttgaaag 480 gaagatattg gggggagggt ctaatggggg gcaattcggc gccccggatc aatttaaact 540 ttggatagag agaagtgcca agttaaaata attgaatcga ttccccgatg acggaacggt 600 attgagtcct gattagggat cggttgcgtg gggggctaaa ttggaaaact tgaaggggtt 660 tttgaaataa caaaattaag ccaaaatttg ggtaaaaaag atgatccccg ggtataaatt 720 ttcaatcatg actgcggtca tctcaatctt cacaagccgc tatacgatac actacccccc 780 aatctaacag tcccttctct tagcattttt caggctgata ggcacccttc cttaactgcg 840 tcat 844 <210> 2 <211> 849 <212> DNA <213> Streptococcus thermophilus LAB kw15 <400> 2 nnnnnnaagg ntttcggatc gtaaagctct gttgtaagtc aagaacgggt gtgagagtgg 60 aaagttcaca ctgtgacggt agcttaccag aaagggacgg ctaactacgt gccagcagcc 120 gcggtaatac gtaggtcccg agcgttgtcc ggatttattg ggcgtaaagc gagcgcaggc 180 ggtttgataa gtctgaagtt aaaggctgtg gctcaaccat agttcgcttt ggaaactgtc 240 aaacttgagt gcagaagggg agagtggaat tccatgtgta gcggtgaaat gcgtagatat 300 atggaggaac accggtggcg aaagcggctc tctggtctgt aactgacgct gaggctcgaa 360 agcgtgggga gcgaacagga ttagtccttg gtggtagtca gcgatttcat tgcggctgcc 420 ccccggtagg ggaagcccgg attaaaaagg tttttcgaat cgtaaagctt ttttgtagtt 480 cagaaagggg tttgagaatt ggaaagtttc ccctgtgacg gtgcttacca aaaggggaag 540 gcaaatacgt gccgcacccg ggtaatccct agggtcgggc tgttcccgga tttttggggg 600 aataaaggag ggcggggggn ntagtattat ttaaaatttt aagggggtgg gacaacacag 660 attttcttgt ggaaccggtc aatattgggt gcaaaagggg gaaaagggga aagtccgtgg 720 ttagccttaa aagtggcgta tattttnggg agagaacccg gtggggaagg cgctcttttg 780 gattatttac ttgatcttaa gggtgaaagg cgggggggca caaaagaaat gattccccgg 840 gtataaaaa 849

Claims (11)

Leuconostoc inhibits growth of Helicobacter pylori mesenteroides ) LAB kw5 strain [KACC91648P]. Leuconostoc mesenteroides ) LAB kw5 strain [KACC91648P] or a culture for inhibiting Helicobacter pylori as an active ingredient. The method of claim 2,
Streptococcus thermophilus ( Streptococcus) to the active ingredient thermophilus ) LAB kw15 strain [KACC91649P] or a composition for inhibiting Helicobacter pylori to add a culture thereof. Leuconostoc mesenteroides ) A formal formulation comprising LAB kw5 strain [KACC91648P] or a culture thereof. The method of claim 4, wherein
Streptococcus thermophilus ( Streptococcus) to the active ingredient thermophilus ) Formal agent adding LAB kw15 strain [KACC91649P] or its culture. Leuconostoc mesenteroides ) probiotic comprising a LAB kw5 strain [KACC91648P] or a culture thereof. The method according to claim 6,
Streptococcus thermophilus ( Streptococcus) to the active ingredient thermophilus ) probiotic to add LAB kw15 strain [KACC91649P] or its culture. Leuconostoc mesenteroides ) A food additive comprising LAB kw5 strain [KACC91648P] or a culture thereof. The method of claim 8,
Streptococcus thermophilus ( Streptococcus) to the active ingredient thermophilus ) Food additive which adds LAB kw15 strain [KACC91649P] or its culture. Leuconostoc mesenteroides ) Agents for the prevention and treatment of gastritis, gastric ulcer or duodenal ulcer comprising LAB kw5 strain [KACC91648P] or a culture thereof. 11. The method of claim 10,
Streptococcus thermophilus ( Streptococcus) to the active ingredient thermophilus ) Agent for the prevention and treatment of gastritis, gastric ulcer or duodenal ulcer, adding LAB kw15 strain [KACC91649P] or a culture thereof.

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