JP2013116089A - Lactobacillus strain having anti-helicobacter pylori activity, anti-helicobacter pylori medicine containing the lactobacillus strain, and food and drink containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide lactobacillus strains with a high bacterial eradication rate to Helicobacter pylori, having anti-Helicobacter pylori activity hard to be restricted in type of use; anti-Helicobacter pylori medicine containing the lactobacillus strains, and foods and drinks containing the same.SOLUTION: The new strains include Lactococcus lactis HA-7 strain, Leuconostoc citreum HA-12 strain, Weissella paramesenteroides HA-819 strain, Leuconostoc mesenteroides HA-863 strain, and Weissella confusa HA-1315 strain, each of them manifests high anti-Helicobacter pylori activity compared with the standard strain of each strain. The anti-Helicobacter pylori medicine is characterized by at least containing a kind of strain selected from the group consisting of the strains. The foods and drinks are characterized by containing at least a kind of strain selected from the group consisting of the strains or the anti-Helicobacter pylori medicine.

Description

  The present invention relates to a lactic acid strain having an anti-Helicobacter pylori effect, an anti-Helicobacter pylori agent containing the lactic acid strain, and a food and drink containing the lactic acid strain.

  Helicobacter pylori is a gram-negative spiral gonococci found specifically in the gastropyloric vestibule discovered by Warren and Marshall in 1983 [Lancet, I.] 1273-1275 (1983)].

  Helicobacter pylori is highly detected from the gastric mucosa of patients such as gastritis and peptic ulcer, and is considered to be strongly involved in the development of these diseases. In recent years, attention has been focused on gastric cancer, and it is known that Helicobacter pylori is the cause of the development of certain types of gastric cancer. More than 70% of Japanese middle-aged and older people are said to be carriers of Helicobacter pylori.

  From such a background, there is an increasing social interest in the eradication and growth inhibition of Helicobacter pylori. For example, drugs are used for sterilization and growth inhibition of Helicobacter pylori. Drugs commonly used for eradication and growth inhibition of Helicobacter pylori are a combination of proton inhibitors and antibiotics (β-lactam antibiotics, tetracycline antibiotics, macrolide antibiotics, new quinolone antibiotics, etc.) There are about three types of drugs. It is known that eradication and growth inhibition of Helicobacter pylori by drugs will have a certain effect on the suppression of gastric cancer.

  JP-A-2001-143 reports that Lactobacillus gasseri belonging to Lactobacillus gasseri inhibits the growth of Helicobacter pylori and is applied to foods for the purpose of eradicating Helicobacter pylori and preventing infection. (Patent Document 1). Lactic acid bacteria are used in various fermented foods around the world, and are very safe bacteria. Lactic acid bacteria are included in pharmaceuticals and foods in the form of live and dead bacteria. Moreover, the form of the fermentation product obtained by fermentation by lactic acid bacteria is included in pharmaceuticals and foods regardless of the presence or absence of lactic acid bacteria. Those that have no anti-Helicobacter pylori effect if they are not live bacteria, those that have anti-Helicobacter pylori effects even if they are dead, and those that have anti-Helicobacter pylori effects that are obtained by fermentation with lactic acid bacteria regardless of the presence or absence of bacteria The form of lactic acid bacteria having an anti-Helicobacter pylori effect varies depending on the individual lactic acid bacteria. It is known that the anti-Helicobacter pylori effect by lactic acid bacteria is caused by, for example, lactic acid produced by lactic acid bacteria or bacteriocin as an antibacterial substance. In order to sterilize Helicobacter pylori, lactic acid bacteria may be used alone (Non-patent Document 1) or lactic acid bacteria and drugs may be used in combination (Non-Patent Document 2).

Japanese Patent Laid-Open No. 2001-143

Boonyaritichaikij S., Kuwabara K., Nagano J., Kobayashi K., Koga Y., Long-term administration of probiotics to asymptomatic pre-school children for either the eradication or the prevention of Helicobacter pylori infection, Helicobacter, 14 (3) , 202-207, June 2009 Shigeru Kamiya, Effect of probiotics on Helicobacter pylori infection, Journal of Intestinal Bacteriology, 24, 1-12, 2010

  However, if Helicobacter pylori is not completely sterilized by the drug, and if it is not completely sterilized by the first drug administration, then the Helicobacter pylori eradication rate will drop significantly even after the second and subsequent drug administration. Therefore, there is a problem that a large amount of medicine must be used over a long period of time. In addition, when a drug is administered a plurality of times, the drug is taken in a large amount for a long period of time, which may cause side effects such as stress on the patient and appearance of resistant bacteria.

  In addition, when the lactic acid bacterium described in Non-Patent Document 1 is used alone, there is a problem that the eradication rate of Helicobacter pylori is 30% or less and the eradication rate is low. Furthermore, the effect cannot be obtained unless the lactic acid bacterium is used as a living bacterium, and there is a problem that the use form of the lactic acid bacterium is restricted.

  Furthermore, lactic acid bacteria having no anti-Helicobacter pylori effect unless they are viable bacteria cannot be used because they are affected by the drug when used in combination with or in combination with the drug. Therefore, there is a problem that the use form of lactic acid bacteria is restricted, for example, it is necessary to use it in a form other than viable bacteria.

  The present invention has been made to solve the above-mentioned problems, and is a lactic acid bacterium having an anti-Helicobacter pylori effect that has a high sterilization rate of Helicobacter pylori and is unlikely to be restricted in use, and an anti-Helicobacter pylori containing the lactic acid bacterium It aims at providing the food-drinks containing an agent and its lactic acid bacteria.

  The strain of the present invention is a strain of Lactococcus lactis HA-7 having a deposit number of NITE P-1157, and Leuconostoc citreum HA-12 having a deposit number of NITE P-1158 (Leuconostoc). citreum HA-12) strain, Weissella paramesenteroides HA-819 (Weissella paramesenteroides HA-819) strain with accession number NITE P-1159, Leuconostoc mesenteroides HA- with accession number NITE P-1160 863 (Leuconostoc mesenteroides HA-863) strain, Weissera konfusa HA-1315 (Wei) whose accession number is NITE P-1161 It is a sella confusa HA-1315) strain.

  The strain of the present invention exhibits a high anti-Helicobacter pylori effect as compared with the standard strain of each strain. Therefore, if the said strain is used, the anti- Helicobacter pylori effect can be improved compared with the case where the standard strain of each strain is used. The anti-Helicobacter pylori effect is at least one of the effect of eradicating Helicobacter pylori and the effect of suppressing proliferation.

The strain of the present invention can be cultured under any conditions according to a conventional method for strain culture. The culture method is not particularly limited, and a conventionally known method can be appropriately selected and used.
Each of the strains of the present invention is preferably isolated from garlic.

The strain of the present invention isolated from garlic has a higher anti-Helicobacter pylori effect than lactic acid strains isolated from other vegetables.
Isolation from garlic can be performed by a general method. For example, after washing garlic with sterilized physiological saline, bacteria attached to garlic can be isolated from the washing solution. In addition, after crushing garlic, add the sample obtained from the pulverized product to the medium for separating lactic acid bacteria containing bromcresol purple, and collect the strains that have turned yellow around the colonies of the strains after culturing each strain. Can also be isolated.

The strain of the present invention isolated from a garlic ground product has a higher anti-Helicobacter pylori effect than a lactic acid strain isolated without pulverizing garlic.
In addition, the strain of the present invention isolated from garlic pulverized product has a higher anti-Helicobacter pylori effect than the standard strain of the strain.

  Moreover, since a strain having resistance to the garlic component extracted from garlic can be obtained by being isolated from garlic, it can be used in combination with a garlic component having an anti-Helicobacter pylori effect.

There are no particular restrictions on the garlic variety or production area used.
The first anti-Helicobacter pylori agent of the present invention is a strain of Lactococcus lactis HA-7 having a deposit number of NITE P-1157, and Leuconostoc having a deposit number of NITE P-1158.・ Cytreum HA-12 (Leuconostoc citreum HA-12) strain, Weissella paramesenteroides HA-819 (Weissella paramesenteroides HA-819) strain with accession number NITE P-1159, accession number is NITE P-1160 Leuconostock mesenteroides HA-863 (Leuconostoc mesenteroides HA-863) strain, Weissera confusa HA with accession number NITE P-1161 It contains at least one strain selected from the group consisting of each of the strains isolated from -1315 (Weissella confusa HA-1315) strain and garlic.

  According to such a first anti-H. Pylori agent, the anti-H. Pylori effect can be improved as compared with the anti-H. Pylori agent using the standard strain of each strain.

  The first Helicobacter pylori agent of the present invention is preferably a strain isolated from garlic. Strains isolated from garlic have a higher anti-Helicobacter pylori effect than lactic acid strains isolated from other vegetables and are resistant to garlic components extracted from garlic, so garlic components extracted from garlic Can be used together.

An anti-Helicobacter pylori agent is a drug having an anti-Helicobacter pylori effect.
Moreover, in the 1st anti- Helicobacter pylori agent of this invention, it is preferable that the strain used is a dead microbe.

  In this way, it can be used for anti-Helicobacter pylori even in forms other than viable bacteria, so there are restrictions on the use form of strains compared to using strains that do not have anti-Helicobacter pylori effects unless they are live bacteria. It can be made difficult to receive.

The dead bacteria are strains obtained by, for example, heat treatment of live bacteria.
In the first anti-Helicobacter pylori agent of the present invention, the culture solution of the strain may be used as a constituent of the anti-Helicobacter pylori agent, or the culture solution sterilized after culturing the strain is used as the first anti-Helicobacter pylori agent. -You may use as a constituent material of a pylori agent. In addition, regardless of whether or not the culture solution is sterilized, the culture solution may be dried and a solid such as a powder may be used as a constituent of the first anti-Helicobacter pylori agent. Since the culture solution contains a strain, the strain is included in the first anti-Helicobacter pylori agent when the culture solution or a solid material obtained by drying the culture solution is used. In addition, regardless of whether the culture solution is sterilized, the culture solution may be separated by centrifugation or the like, and the solid content containing the strain or the like may be used as a constituent of the first anti-Helicobacter pylori agent.

The second anti-Helicobacter pylori agent of the present invention contains as an active ingredient a fermentation component obtained by fermentation with at least one strain selected from the group consisting of the strains described above.
A fermentation component is a component obtained by fermentation of each strain. This component is contained, for example, in the supernatant of the culture solution of the strain. The supernatant is a liquid portion obtained by culturing the strain, separating the cultured medium after culturing by centrifugation or the like, and precipitating solids such as bacteria.

In the second anti-Helicobacter pylori agent of the present invention, the culture solution of the strain may be used as a constituent of the second anti-Helicobacter pylori agent.
Since the culture solution contains fermentation components obtained by fermentation of the strain, the fermentation components obtained by fermentation by the strain are included in the second anti-Helicobacter pylori agent even if the culture solution is used. .

  The first and second anti-Helicobacter pylori agents of the present invention may contain excipients, binders, disintegrants, lubricants, coating agents and the like, respectively. In addition, additives such as diluents, buffers, suspending agents, emulsifiers and the like that can be usually used in the technical field of pharmaceutical production may be contained. Moreover, you may mix | blend another chemical | medical agent as needed.

  The excipient is not particularly limited. For example, sugars such as sucrose, lactose, mannitol and glucose, starches such as corn starch, potato starch and rice starch, inorganic substances such as calcium carbonate and calcium sulfate, crystalline cellulose, distillation Examples thereof include water, purified water, sesame oil, soybean oil, corn oil, olive oil, and cottonseed oil.

  Examples of the binder include, but are not limited to, for example, chitosan, dextrin, sodium alginate, carrageenan, guar gum, gum arabic, agar and other polysaccharides, tragacanth, gelatin, gluten and other natural polymers, hydroxypropylcellulose, methylcellulose, Examples include cellulose derivatives such as hydroxypropylmethylcellulose, ethylcellulose, hydroxypropylethylcellulose, sodium carboxymethylcellulose, synthetic polymers such as polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl acetate, polyethylene glycol, polyacrylic acid, polymethacrylic acid, and vinyl acetate resin. Can do.

  The disintegrant is not particularly limited. For example, cellulose derivatives such as carboxymethyl cellulose, carboxymethyl cellulose calcium, low-substituted hydroxypropyl cellulose, sodium carboxymethyl starch, hydroxypropyl starch, corn starch, potato starch, rice starch, and partial α And starches such as modified starch.

  The lubricant is not particularly limited, and examples thereof include talc, stearic acid, calcium stearate, magnesium stearate, colloidal silica, hydrous silicon dioxide, waxes, and hardened oil.

  The coating agent is not particularly limited. For example, dimethylaminoethyl methacrylate / methacrylic acid copolymer, polyvinyl acetal diethylaminoacetate, ethyl acrylate / methacrylic acid copolymer, ethyl acrylate / methyl methacrylate / Water-insoluble polymers such as methacrylated trimethylammonium ethyl copolymer, ethyl cellulose, methacrylic acid / ethyl acrylate copolymer, enteric polymers such as hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, methylcellulose, Examples thereof include water-soluble polymers such as hydroxypropylmethylcellulose, polyvinylpyrrolidone and polyethylene glycol.

  The administration form is not limited, and may be administered orally, for example, in the form of powder, granules, tablets, capsules, syrups, etc., or administered parenterally such as injections. It may be a thing.

Each of the first and second anti-Helicobacter pylori agents of the present invention preferably contains a garlic component extracted from garlic.
Some garlic components extracted from garlic have an anti-Helicobacter pylori effect. Therefore, in the first anti-Helicobacter pylori agent, the anti-Helicobacter pylori effect can be enhanced by including a garlic component having an anti-Helicobacter pylori effect as compared with the case where the strain is used alone. In addition, in the second anti-Helicobacter pylori agent, by including a garlic component having an anti-Helicobacter pylori effect, compared to the case where a fermentation component obtained by fermentation with a strain is used alone as an active ingredient, Anti-Helicobacter pylori effect can be enhanced.

  Moreover, since the garlic component extracted from garlic is a natural component, side effects caused by the use of a non-natural component such as a chemical agent can be prevented, so the first and second anti-Helicobacter pylori agents Each safety can be improved.

In the first anti-Helicobacter pylori agent, examples of the strain used when it contains a garlic component include strains isolated from garlic.
When the second anti-Helicobacter pylori agent contains a garlic component, a fermentation component obtained by fermentation with a strain isolated from garlic can be used.

Moreover, examples of the garlic component extracted from garlic include allicin and ajoene, which are antibacterial substances.
The 1st food / beverage products of this invention are characterized by including the at least 1 sort (s) of strain selected from the group which consists of said strain.

  Such foods and drinks contain the above-described strains having a high anti-Helicobacter pylori effect. Therefore, according to such food and drink, the anti-Helicobacter pylori effect can be improved.

  As the first food and drink, solid foods and beverages in any form such as powder, granule, lump, etc. by mixing the above-mentioned strain directly with any food or mixing with additives such as fragrance The thing made into food is mentioned. For example, the above-described strains are added to dairy products and vegetables to make fermented foods.

In addition, there are products obtained by mixing arbitrary excipients, binders, diluents, and the like into health foods of any form such as powders, granules, tablets, capsules, and beverages.
The 2nd food / beverage products of this invention are characterized by including the above-mentioned anti- Helicobacter pylori agent.

According to such food and drink, the anti-Helicobacter pylori agent described above is included, so that the anti-Helicobacter pylori effect can be improved.
As 2nd food / beverage products, the solid food, drink food, etc. similar to 1st food / beverage products are mentioned.

  Also, as in the first food and drink, any excipients, binders, diluents and the like are mixed to form health foods of any form such as powders, granules, tablets, capsules, beverages, etc. There is also.

Next, an embodiment of the present invention will be described with an example. However, the present invention is not limited to these examples.
1. Isolation of HA-7, HA-12, HA-819, HA-863, HA-1315 strains exhibiting an anti-Helicobacter pylori effect (1) Garlic (white species) commercially available at supermarkets was used. After removing the garlic crust, it was pulverized in a food processor. After holding for 1 hour, 1 g of this pulverized product was collected and suspended in 10 ml of sterilized physiological saline, and 0.1 ml was inoculated into BCP-added plate count agar (manufactured by Nissui Pharmaceutical). This was designated as agar medium 1.
(2) The above (1) was repeated, and 2000 strains of colonies with yellowing around the colonies were collected from the agar medium 1. Since the periphery of the colony of the lactic acid strain resistant to the garlic component is yellowed, a lactic acid strain resistant to the garlic component can be obtained by collecting the colony. Each of the collected 2000 strains was cultured in an MRS liquid medium (Difco).
(3) Helicobacter pylori suppression test was performed for each strain. The method of Helicobacter pylori suppression test is described below. About each strain, after culture | cultivating until the turbidity of a culture solution will be set to 1.0, culture | cultivation 0.25mL is extract | collected and culture which contains 10 ⁷ colony forming units (colon forming units: cfu) of Helicobacter pylori ATCC43504 strain. Mixed with 0.25 mL of liquid. After maintaining at 37 ° C. for 1 hour, the solution was appropriately diluted and cultured on a Helicobacter agar medium (manufactured by Nissui Pharmaceutical) at 37 ° C. for 7 days under microaerobic conditions, and the viable count of Helicobacter pylori was measured. Helicobacter pylori ATCC 43504 strain used was cultured in Brucella broth (manufactured by Difco) containing 10% horse serum.

  As a result, it was shown that the viable count of Helicobacter pylori is significantly reduced compared to a blank containing no strain culture medium when 5 specific strains were used among the 2000 strains collected. It was revealed that there is a high anti-Helicobacter pylori effect. Hereinafter, these five strains are referred to as HA-7, HA-12, HA-819, HA-863, and HA-1315.

  Table 1 shows the results of the Helicobacter pylori suppression test of HA-7, HA-12, HA-819, HA-863, and HA-1315.

Note that HA-7, HA-12, HA-819, HA-863, and HA-1315 are deposited with the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology. HA-7 has accession number NITE P-1157, HA-12 has accession number NITE P-1158, HA-819 has accession number NITE P-1159, and HA-863 has accession number NITE P-1160. HA-1315 has been deposited under the deposit number NITE P-1161.
(4) Lactic acid strain isolated from garlic ground product, lactic acid strain isolated from garlic soaked product, lactic acid strain isolated from cabbage, Helicobacter pylori suppression test for each, In the above (1), garlic ground The thing was used. The Helicobacter pylori suppression test for the samples 1a to 10a, 1b to 10b, and 1c to 10c described below that the lactic acid strain isolated from the garlic pulverized product as described above has a high anti-Helicobacter pylori effect. Made clear by doing.

  Samples 1a to 10a are lactic acid bacterial strains isolated from garlic ground. Samples 1a-10a were obtained using the method described next. From the agar medium 1 described in the above (1), 100 colonies in which the periphery of the colony turned yellow were collected. The collected colonies of 100 strains were divided into 10 groups of 10 strains for each. Each of the 10 groups was cultured in MRS liquid medium (Difco). 0.1 mL of each culture solution having a turbidity of 1.0 was collected from each group and collected. This was used as a sample for Helicobacter pylori suppression test. Ten samples were prepared and designated as 1a to 10a.

  Samples 1b to 10b are lactic acid strains isolated from garlic soup. Samples 1b-10b were obtained using the method described below. The skin of garlic (white seed) was removed, and 1 piece of garlic bulb (about 4 g) was immersed in 50 ml of sterilized physiological saline as it was. After holding for 1 hour, 0.1 ml of physiological saline was collected and inoculated into a BCP-added plate count agar. This was designated as agar medium 2, and samples 1b to 10b were obtained using the same method as the method for obtaining samples 1a to 10a.

  Samples 1b to 10b are lactic acid strains isolated from cabbage. Samples 1c-10c were obtained using the method described below. The cabbage used in the supermarket was used. After removing a few cabbage shells, immerse 4 to 10 g of cabbage in 50 ml of sterilized physiological saline, crush it with a food processor, hold it for 1 hour, collect 0.1 ml, and add BCP plate count agar Inoculated. This was used as agar medium 3, and samples 1c to 10c were obtained using the same method as the method for obtaining samples 1a to 10a.

  For each sample, the Helicobacter pylori suppression test is performed as described above in 1. This was carried out using the method described in (3). The results are shown in Table 2.

As shown in Table 2, the samples of 1a to 10a exhibited a high anti-Helicobacter pylori effect as compared with the samples of 1b to 10b and 1c to 10c. In addition, from the results of lactic acid strains 1a to 10a and 1b to 10b isolated from garlic, it is more effective to use garlic after crushing garlic than to use it without crushing garlic.・ It was revealed that the H. pylori effect is high.
2. Identification of isolated strain (1) HA-7 was subjected to 16S rDNA (16S ribosomal DNA) analysis using a known method. As a result, HA-7 showed high homology in the partial sequence of 16S rDNA with the strain of Lactococcus lactis already registered in the database. From this, it was found that HA-7 is Lactococcus lactis. Similarly, 16S rDNA analysis was performed on HA-12, HA-819, HA-863, and HA-1315. As a result, HA-12 is Leuconostoc citreum, HA-819 is Weicella Paramesenteroides, HA-863 is Leuconostoc mesenteroides, and HA-1315 is Weicera. I found out that it was a confetti. Moreover, since these strains were not 100% homologous with the reference strain, it was found that these strains were novel strains different from known strains.
(2) The mycological properties (morphological characteristics, reproductive mode, physiological / chemical characteristics, etc.) of HA-7, HA-12, HA-819, HA-863, and HA-1315 were analyzed. Sugar assimilation was analyzed using API (API) CH kit (Sysmex Biomelieu). The analysis results are shown in Table 3.

  The bacteriological properties of HA-7, HA-12, HA-819, HA-863, and HA-1315 are different in the assimilation ability of sugar as described below compared to the respective reference strains. It has been shown. The HA-7 strain did not show assimilation of D mannitol, but HA-7 showed assimilation against D mannitol. The HA-12 reference strain did not show assimilation of D-xylose, but HA-12 showed assimilation against D-xylose. The HA-819 reference strain did not show assimilation of D-xylose, but HA-819 showed assimilation against D-xylose. HA-863 showed weak assimilation against gentiobiose compared to its reference strain. HA-1315 showed weak assimilability for D-galactose compared to its reference strain. From the above, it was found that these strains are novel strains different from known strains.

Hereinafter, the above 1. HA-7, HA-12, HA-819, HA-863, and HA-1315 isolated from garlic described in 1. are collectively referred to as strains isolated from garlic.
The blanks in Tables 4 and 6 to 8 are mixed with a culture solution containing 10 ⁵ cfu / ml of each strain and physiological saline at a ratio of 1: 1 and kept at 37 ° C. for 1 hour, and then mixed. The result of inoculating the solution into a BCP-added plate count agar and counting the number of colonies of each strain is shown.
3. Helicobacter pylori suppression test Helicobacter pylori suppression test was performed for each strain isolated from garlic and each reference strain. The test is: The same method as described in (3) was performed. The results are shown in Table 4.

It was clarified that the strain isolated from garlic has a high anti-Helicobacter pylori effect by reducing the viable count of Helicobacter pylori compared to the reference strain.
4). Resistance test against garlic component Each strain isolated from garlic and each reference strain were subjected to a resistance test against garlic component. Garlic juice and ajoen were used for the resistance test. Ajoene is an antibacterial substance derived from garlic. Ajoene was produced and analyzed according to the method disclosed in Japanese Patent No. 2608252.

Resistance tests against garlic juice were performed as described below. After immersing 100 g of garlic from which the outer skin had been removed in 70% ethanol, 20 ml of sterilized water was added and treated with a food processor to obtain garlic juice. A culture solution containing 10 ⁵ cfu / ml of each strain and squeezed garlic were mixed at a ratio of 1: 1 and held at 37 ° C. for 1 hour, and then the mixture was inoculated into a BCP-added plate count agar. The number of colonies of each strain was counted.

Resistance tests against ajoene were performed as described below. The culture solution containing 10 ⁵ cfu / ml of each strain was added so as to contain 150 μg / ml of ajoene, and held for 1 hour. The results are shown in Table 5.

As shown in Table 5, when the reference strain was used, the viable count of each strain was greatly reduced, but the strain isolated from garlic contained garlic juice and ajoene in the culture solution. However, the viable count of each strain did not decrease. Therefore, it was revealed that the strain isolated from garlic has resistance to the garlic component.
5. First anti-Helicobacter pylori agent For each strain isolated from garlic, a first anti-Helicobacter pylori agent containing dead strains of each strain was prepared as described below.

  About each culture solution of the strain isolated from the garlic, it culture | cultivated until turbidity became 1.0, and the culture solution was heat-processed on 121 degreeC and the conditions for 15 minutes. This culture solution was used as the first anti-Helicobacter pylori agent.

About the 1st anti- Helicobacter pylori agent, the Helicobacter pylori suppression test was done.
The culture solution of Helicobacter pylori ATCC 43504 strain was cultured in Brucella broth containing 10% horse serum to 10 ⁷ cfu / ml.

  The first anti-Helicobacter pylori agent and the above Helicobacter pylori ATCC 43504 culture solution are mixed, incubated at 37 ° C. for 1 hour, diluted appropriately, and applied to Helicobacter agar medium (manufactured by Nissui Pharmaceutical). After culturing at 37 ° C. for 7 days under aerobic conditions, the number of Helicobacter pylori colonies was counted. The results are shown in Table 6.

When the first anti-Helicobacter pylori agent was used, the number of Helicobacter pylori was decreased compared to the blank, and thus a high anti-Helicobacter pylori effect was recognized. Therefore, since it can be used in the form of dead bacteria by heat-treating the strain instead of viable bacteria, it can be made difficult to be restricted by the usage form of the strain.
6). Second anti-Helicobacter pylori agent For each strain isolated from garlic, a second anti-Helicobacter pylori agent was produced as described below, using as an active ingredient a fermentation component obtained by fermentation with the respective strain. .

  Culturing the culture solution of each strain isolated from garlic until the turbidity becomes 1.0, centrifuging the culture solution, and filtering the supernatant with a 0.45 μm sterilized filter. I got Qing. This supernatant was used as a second anti-Helicobacter pylori agent.

A Helicobacter pylori suppression test was performed on the second anti-Helicobacter pylori agent.
As the culture solution of Helicobacter pylori ATCC 43504, a culture solution similar to that described in 5 above was used.

  250 μl of the second anti-Helicobacter pylori agent and Helicobacter pylori ATCC 43504 culture solution were mixed at a time, incubated at 37 ° C. for 1 hour, diluted as appropriate, and applied to Helicobacter agar medium under microaerobic conditions. After culturing at 0 ° C. for 7 days, the number of Helicobacter pylori colonies was counted. The results are shown in Table 7.

When the second anti-Helicobacter pylori agent was used, since the number of Helicobacter pylori colonies was reduced as compared with the blank, it was confirmed that there was a high anti-Helicobacter pylori effect.
7). Anti-Helicobacter pylori containing garlic component Anti-Helicobacter pylori containing each strain isolated from garlic and ajoene having anti-Helicobacter pylori effect as a garlic component was prepared as described below.

Each strain isolated from garlic was cultured until the turbidity reached 1.0. Ajoene was added to the culture solution to prepare an anti-Helicobacter pylori agent containing a garlic component.
A Helicobacter pylori suppression test was conducted for anti-Helicobacter pylori agents containing garlic components.

As the culture solution of Helicobacter pylori ATCC 43504, a culture solution similar to that described in 5 above was used.
250 μl of the culture solution of Helicobacter pylori ATCC 43504 strain was mixed with an anti-Helicobacter pylori agent containing a garlic component. (Ajoene was added at 15 μg / ml.) After incubation at 37 ° C. for 1 hour, the cells were cultured on Helicobacter agar medium at 37 ° C. for 7 days under microaerobic conditions, and the number of Helicobacter pylori colonies was counted. Helicobacter pylori suppression test was also performed when no ajoene was added. The results are shown in Table 8.

The anti-Helicobacter pylori agent of the present invention reduces the number of Helicobacter pylori colonies by using a strain isolated from garlic in combination with ajoen, compared to the case where each strain is used alone. From this, it was revealed that the anti-Helicobacter pylori effect increases. It was also confirmed that the strain isolated from garlic was alive in the presence of ajoene.
8). Food / beverage products containing the strain of the present invention and anti-Helicobacter pylori The fermented milk and yogurt can also be produced by adding the strain of the present invention and the anti-Helicobacter pylori to raw milk of livestock such as cattle. Moreover, it can also be made into fermented food by adding to the squeeze or paste of vegetables and fruits. Furthermore, arbitrary excipient | filler, binder, a diluent, etc. can be mixed and processed into the health food of the form of a powder, a granule, a tablet, a capsule, a drink.

Since the food / beverage products of this invention contain the strain of this invention and an anti- Helicobacter pylori agent, there exists a high anti- Helicobacter pylori effect compared with the food / beverage products which do not contain these.
As described above, the strain isolated from garlic has a high anti-Helicobacter pylori effect. Moreover, since it has tolerance with respect to the antibacterial component derived from a garlic, it can also be used together with these antibacterial components. Furthermore, the use form of the strain is not easily restricted, and a high anti-Helicobacter pylori effect can be achieved using fermentation components obtained by fermentation of live bacteria, dead bacteria, and strains.

  In addition, this invention is not limited to the said embodiment at all, and it cannot be overemphasized that it can implement with a various aspect in the range which does not deviate from this invention.

Claims (12)

  Lactococcus lactis HA-7 NITE P-1157 strain.   Leuconostoc citreum HA-12 NITE P-1158 strain.   Weisella paramesenteroides HA-819 (Weissella paramesenteroides HA-819) NITE P-1159 strain.   Leuconostoc mesenteroides HA-863 (Leuconostoc mesenteroides HA-863) NITE P-1160 strain.   Weissella confusa HA-1315 (Weissella confusa HA-1315) NITE P-1161 strain.   The strain according to any one of claims 1 to 5, wherein the strain is isolated from garlic.   An anti-Helicobacter pylori agent comprising at least one strain selected from the group consisting of the strains according to claim 1.   The anti-Helicobacter biloli agent according to claim 7, wherein the strain is killed.   An anti-Helicobacter pylori agent comprising as an active ingredient a fermentation component obtained by fermentation with at least one strain selected from the group consisting of the strains according to claim 1.   The anti-Helicobacter pylori agent according to any one of claims 7 to 9, comprising a garlic component extracted from garlic.   A food or drink comprising at least one strain selected from the group consisting of the strains according to claim 1.   Food-drinks characterized by including the anti- Helicobacter pylori agent as described in any one of Claims 7-10.