CN1514875A - Kimchi lactic acid bacteria that inhibit the growth of Helicobacter pylori and a highly functional food that prevents gastrointestinal disorders - Google Patents

Abstract

The present invention relates to Kimchi lactic acid bacteria that inhibit growth of Helicobactor pylori, and foods comprising the same, and particularly to a gastrointestinal-tract-protecting high functional food comprising Kimchi lactic acid bacteria that inhibits attachment of Helicobactor pylori to the gastrointestinal tract, and inhibits growth thereof. The Kimchi lactic acid bacteria of the present invention inhibits attachment of Helicobactor pylori that inhabit the gastric mucous membrane tissue of the gastrointestinal tract, and inhibits growth thereof, and thus are very effective for Helicobactor pylori extermination and prevention of reinfection therefrom. In addition, a gastrointestinal-tract-protecting high functional food comprising the Kimchi lacti acid bacteria is very effective for preventing and treating gastritis, peptic ulcers, and duodenal ulcers caused by Helicobactor pylori.

Description

Kimchi lactic acid bacteria that inhibit the growth of Helicobacter pylori and a highly functional food that prevents gastrointestinal disorders

technical field

The present invention relates to kimchi lactic acid bacteria capable of inhibiting the growth of Helicobacter pylori (Helicobacter pylori), and a high-function food capable of protecting the gastrointestinal tract, in particular to a high-function food containing Gastrointestinal gastric mucosal tissue and inhibit the growth of kimchi lactic acid bacteria, so it can effectively prevent and treat gastritis, gastric ulcer and duodenal ulcer.

Background technique

Gastrointestinal diseases are the most widely occurring diseases in the world, and their therapeutic drugs form the largest single market (antiulcer drugs: US$ 15.8 billion (1999)).

In 1983, microbiologists Marchall and Warren first discovered ichthyosis bacteria in the gastric mucosal biopsy of B-type gastritis patients, cultured the bacteria and found that it was a related species of Campylobacter genus new strains, and since then, research surrounding H. pylori and diseases of the upper gastrointestinal tract such as gastritis and gastric ulcers has become increasingly active. Helicobacter pylori is a Gram-negative bacillus surrounded by flagella formed by a thin membrane, which can secrete a large amount of urease and spread through the excrement-oral route. Since it was confirmed by the World Health Organization (WHO) in 1994 As a carcinogen, it has attracted worldwide attention.

It is known that the pathogenic mechanism of Helicobacter pylori is: bacteria enter the gastric cavity through food, penetrate the gastric mucosa with high-speed swimming flagella, and secrete a large amount of urease, which raises the pH value of the mucosa to an alkaline state. Helicobacter pylori parasitizes the area between the upper layer of gastric epithelial cells and gastric mucosal cells, causing abnormal secretion of gastrin, which in turn leads to a large amount of gastric acid, eventually forming inflammation and ulcers. Therefore, according to the latest reports, Helicobacter pylori is an important factor causing gastric adenocarcinoma, and the WHO recognizes Helicobacter pylori as a type I carcinogen.

H. pylori infection has been reported to be particularly prevalent in developing countries, including South Korea. According to the report of Paek et al., nearly 80% or more Koreans are suspected of being infected with Helicobacter pylori (Paek Seungchul et al., Korean Microbial Society Paper, 25:45-52, 1990). Therefore, there is an urgent need to develop a method for treating and preventing H. pylori infection.

Chemotherapy with antibiotics is currently the most commonly used method for treating microbial infection diseases. Although this method is effective in the short term, it is worrying that drug-resistant strains of microorganisms may develop.

In addition, Rauws et al. administered bismuth preparations (bismuth preparations), amoxicillin (amoxicillin) and metronidazole (metronidazole) three antibacterial drugs at the same time, and carried out antibacterial treatment against Helicobacter pylori, and obtained better results (Rauws et al. EAJ, Langenberg W, Houthoff JH, Zanen HC and Tytgat GHJ, Gastroenterol, 94:33-40, 1988). Borody et al. applied bismuth preparations, tetracycline (tetracycline) and metronidazole to patients with gastric ulcer at the same time, and the reported effective rate was about 80% (Borody T, Lenne J and Moore-Jones D, Gastroenterol, 98: A24, 1990 ), so the application of these three antibiotics has become the most commonly used method for the treatment of Helicobacter pylori infection. However, various side effects have been reported, such as diarrhea, reflux esophagitis, suppression of normal intestinal flora, and development of resistant strains due to continuous antibiotic treatment.

Recently, the function of lactic acid bacterial fermented materials in the prevention and treatment of gastrointestinal diseases has attracted increasing attention, and many studies are also underway on probiotic Lactobacillus, which is able to inhibit the growth of human pathogenic bacteria. Lactic acid bacteria are fermentation products that produce bacteriocin-related antibiotic raw materials as well as lactic acid, acetic acid, and hydrogen peroxide. Marie-Helen Coconier et al. have cultivated a human Lactobacillus acidophilus strain LB, which can produce antibiotics against Helicobacter pylori in vitro, and it has been confirmed in vitro that its culture supernatant can inhibit the attachment of Helicobacter pylori to humans. The gastrointestinal cells can also prevent urease activation and inhibit feline Helicobacter pylori (H.felis) infection in mice (Marie-HelenCoconier et al., Environ.Microbiol, November, 4573-4580, 1998).

In addition, Yuji Aida M.T. et al. have confirmed that Helicobacter pylori infection is inhibited in germ-free mice orally administered with Lactobacillus salivarius. U.S. Patent No. 5,578,392 describes the inhibition of Lactobacillus johnsonii (L.johnsonii) Function of Helicobacter pylori attachment to gastrointestinal cells.

In addition, according to the prescription used by Canducci et al., the culture solution of inactivated human Lactobacillus acidophilus was used, and the antibiotics raveprazol, clarithromycin (clarithromycin) and amoxicillin were used for treatment at the same time. As a result, the killing rate of Helicobacter pylori was increased ( Canducci F. et al., Aliment Pharmacol. Ther, 14:1625-1629, 2000).

Similarly, Korean Patent Application No. 1999-0040387 discloses an antibody against Helicobacter pylori, and a protein containing Lactococcus sp. HY 49, Lactobacillus Casei HY 2782, bifidobacterium longum Longum) HY 8001 and other foods have shown the function of inhibiting Helicobacter pylori.

Therefore, materials and methods capable of preventing Helicobacter pylori from attaching to the gastrointestinal tract and inhibiting its growth are required, and foods and medicines containing these materials have been developed.

Contents of the invention

The object of the present invention is to provide a high-function kimchi lactic acid bacterium that protects the gastrointestinal tract, which can inhibit the growth of Helicobacter pylori and inhibit its attachment to the gastrointestinal tract. The kimchi lactic acid bacterium is isolated from kimchi, It is the main source of supply of lactic acid bacteria for Koreans and endows traditional foods with beneficial functions.

Another object of the present invention is to provide a high-function food for protecting the gastrointestinal tract, comprising kimchi lactic acid bacteria which can effectively prevent and treat gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori.

Another object of the present invention is to provide a high-function kimchi that protects the gastrointestinal tract, including kimchi lactic acid bacteria that can effectively prevent and treat gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori.

Another object of the present invention is to provide a medicine capable of preventing and treating gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori, comprising kimchi lactic acid bacteria.

In order to achieve these objects, the present invention provides Kimchi lactic acid bacteria capable of inhibiting the growth of Helicobacter pylori and inhibiting its attachment to the gastrointestinal tract.

The invention also provides a high-function food for protecting the gastrointestinal tract containing kimchi lactic acid bacteria, which can inhibit the growth of Helicobacter pylori, and can effectively prevent and treat gastritis, gastric ulcer and duodenal ulcer.

The present invention also provides a high-function kimchi containing kimchi lactic acid bacteria that protects the gastrointestinal tract, can inhibit the growth of Helicobacter pylori, and can effectively prevent and treat gastritis, gastric ulcer and duodenal ulcer.

The present invention also provides a medicine capable of preventing and treating gastritis, gastric ulcer and duodenal ulcer, comprising Kimchi lactic acid bacteria capable of inhibiting the growth of Helicobacter pylori.

Detailed ways

The present invention will be explained in detail below.

Since the ingredients of kimchi are cabbage and radish, etc., and contain many lactic acid bacteria, kimchi has many useful functions, including anti-mutation and anti-cancer. The microorganisms involved in kimchi fermentation include L. plantarum, L. brevis, Lactococcus, Leuconostoc, Streptococcus , Pediococcus (Pediococcus), etc., with the different fermentation stages, the types and component ratios of bacteria in the total microbial flora formed are also different.

In a study on Kimchi lactic acid bacteria that inhibit the growth of Helicobacter pylori, the inventors of the present invention confirmed that among the microorganisms involved in the kimchi fermentation process, the Kimchi lactic acid bacteria that inhibited the growth of Helicobacter pylori also inhibited its attachment to the gastrointestinal tract Road, thereby perfecting the present invention.

The present invention provides kimchi lactic acid bacteria that inhibit the growth of Helicobacter pylori and inhibit its attachment to the gastrointestinal tract.

The preferred isolation and identification conditions for kimchi lactic acid bacteria are: ferment kimchi by conventional methods, store at 1-40°C for 90 days or its pH value reaches about 3.0; more preferred conditions are: ripen at 1-25°C for 90 days, or Aging at 30-40°C for 30 days.

Lactic acid bacteria are isolated in this way: samples are extracted from kimchi made by conventional methods, cultivated on MRS agar medium, and 588 kimchi lactic acid bacteria that can inhibit the growth of Helicobacter pylori are selected, and then selected to inhibit the growth of Helicobacter pylori. Kimchi lactic acid bacteria in the gastrointestinal tract.

The kimchi lactic acid bacteria are preferably found among the lactic acid bacteria produced during the kimchi fermentation process, Leuconostoc citrophila and Lactobacillus species, more preferably, they are selected from the group consisting of Leuconostoc citrophila EUG4427 (KCCM-10274), Lactobacillus One or more of the group consisting of EUG6017 (KCCM-10273), Lactobacillus EUG4066 (KCCM-10270), Lactobacillus EUG4048 (KCCM-10271) and Lactobacillus EUG6404 (KCCM-10272).

Leuconostoc or Lactobacillus isolated from kimchi can inhibit the growth of Helicobacter pylori parasitic in gastric mucosal tissue and inhibit its attachment to the gastrointestinal tract, which is effective for killing Helicobacter pylori and It is very effective in preventing reinfection.

At the same time, the lactic acid bacteria that function here are not derived from human intestines or milk fermentation products, but from kimchi, which is the main source of lactic acid bacteria for Koreans. This function can be obtained simply by adding bacteria to traditional kimchi and other foods, and the supply source of functional lactic acid bacteria can also be diversified. Therefore, the present invention also provides a high-function food for protecting the gastrointestinal tract, comprising Leuconostococcus citophilus species or Lactobacillus species obtained from kimchi, which is effective in the prevention and treatment of gastritis, gastric ulcer and Duodenal ulcer.

The functional food containing kimchi lactic acid bacteria and capable of inhibiting the growth and attachment of Helicobacter pylori to the gastrointestinal tract can be a starter such as a culture broth, or can be freeze-dried lactic acid bacteria powder added to various foods such as: beverages, fermented milk, Tofu, fermented soybean paste, bean-paste soup obtained by grinding fermented soybeans, jelly, etc. Moreover, on the premise that the function of lactic acid bacteria is not inhibited, the preparation methods of these foods are not limited, and other commonly used food additives can also be added according to nutritional needs and consumer preferences.

The present invention also provides a high-function pickle for protecting the gastrointestinal tract, which contains Leuconostococcus citricophilus species or Lactobacillus species obtained from the pickle, which can effectively prevent and treat gastritis, gastric ulcer caused by Helicobacter pylori. Ulcers and duodenal ulcers.

Kimchi containing kimchi lactic acid bacteria that can inhibit the growth of Helicobacter pylori and adhere to the gastrointestinal tract can be prepared by adding kimchi lactic acid bacteria culture fluid or freeze-dried lactic acid bacteria powder as a starter to the kimchi seasoning, and preferably adding frozen Dried lactic acid bacteria powder to maintain kimchi quality.

The preparation conditions such as temperature, pH value, process method, etc. of kimchi with high function of gastrointestinal protection containing kimchi lactic acid bacteria are not particularly limited under the condition that the function of lactic acid bacteria is not inhibited, and according to the nutritional needs and the needs of consumers Other commonly used food additives can also be added according to preferences, etc.

The present invention also provides a medicament for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori, comprising Leuconostoc citricophilus spp. or Lactobacillus spp. obtained from kimchi . The drug can be prepared as gel, suspension, spray, tablet or liquid.

The present invention will be further described in detail in the examples provided below, however, the following examples are only used to illustrate the present invention, not to limit the present invention.

Example:

Embodiment 1: Isolate the kimchi lactic acid bacteria that can inhibit the growth of Helicobacter pylori

In order to select lactic acid bacteria capable of inhibiting the growth of Helicobacter pylori from Kimchi lactic acid bacteria in vitro, the Kimchi lactic acid bacteria were first isolated, and then the lactic acid bacteria capable of inhibiting Helicobacter pylori were selected therefrom.

(Isolation of Kimchi Lactic Acid Bacteria)

Prepare kimchi using a conventional method, from kimchi stored at 1°C, 4°C, 8°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C at a rate of 24 hours. Or take samples at intervals of 48 hours, and continue for 90 days or until the pH reaches about 3.0 (curing at 1-25°C for 90 days, and at 30-40°C for 30 days). Samples extracted from Kimchi were cultured on MRS agar medium at 37° C. for 72 hours.

(isolation of lactic acid bacteria that inhibit the growth of Helicobacter pylori)

The Helicobacter pylori suspension was spread on Brucell agar containing 10% horse serum and dried for 5 minutes, and then nearly 48,000 lactic acid cultured on MRS agar medium were transferred by tooth-picking method. Bacterial colonies were transferred to Brucella agar coated with Helicobacter pylori suspension, cultured at 37°C for 72 hours under 10% _CO2 partial pressure, and 588 lactic acid bacteria colonies forming transparent rings were selected.

Example 2 Selection of Lactic Acid Bacteria Capable of Inhibiting Helicobacter pylori Attachment to Cell Lines

(selection for the first time)

For the 588 kinds of kimchi lactic acid bacteria selected in Example 1 that can inhibit the growth of Helicobacter pylori in vitro, their activity of inhibiting Helicobacter pylori from attaching to Kato III gastric cancer cells was studied, and lactic acid bacteria with inhibitory ability were selected.

Helicobacter pylori #26695 isolated from the stomach of a Korean man suffering from gastritis and gastric ulcer was obtained from the College of Pharmacy, Kyungsang University; human Kato III gastric cancer cells were obtained from the Korean Cell Line Bank.

First, by measuring the absorbance (A625) at 625nm of the color urease reaction solution (an indicator for measuring the number of Helicobacter pylori), the number of Helicobacter pylori attached to the cells is obtained, and the Helicobacter pylori that can be attached is selected. The number of lactic acid bacteria was reduced to 10 ⁶ or less, and as a result, 70 kinds of lactic acid bacteria were obtained as shown in Table 1 (if the value of A625 was 1.7 or less, the count of Helicobacter pylori was 10 ⁶ or less). Bacteria name The number of attached Helicobacter pylori (inhibiting attachment ability<10 ⁶ ) Bacteria name The number of attached Helicobacter pylori (inhibiting attachment ability<10 ⁶ ) 1 Eug 12142 o 36 Eug 44251 o 2 Eug 6013 o 37 Eug 2253 o 3 Eug 6014 o 38 Eug 2613 o 4 Eug 6017 o 39 Eug 14414 o 5 Eug 0113 o 40 Eug 4416 o 6 Eug 0141 o 41 Eug 4417 o 7 Eug 0340 o 42 Eug 4418 o 8 Eug 0352 o 43 Eug 4419 o 9 Eug 0358 o 44 Eug 44115 o 10 Eug 0445 o 45 Eug 4118 o 11 Eug 0442 o 46 Eug 4427 o 12 Eug 4547 o 47 Eug 4439 o 13 Eug 8126 o 48 Eug 14449 o 14 Eug 8130 o 49 Eug 4410 o 15 Eug 4842 o 50 Eug 4414 o 16 Eug 4848 o 51 Eug 24013 o 17 Eug 8540 o 52 Eug 0618 o 18 Eug 8553 o 53 Eug 40312 o 19 Eug 9617 o 54 Eug 0317 o 20 Eug 6322 o 55 Eug 0315 o twenty one Eug 6713 o 56 Eug 4034 o twenty two Eug 6123 o 57 Eug 40410 o twenty three Eug 6416 o 58 Eug 0412 o twenty four Eug 20553 o 59 Eug 6404 o 25 Eug 14435 o 60 Eug 4047 o 26 Eug 16811 o 61 Eug 4048 o 27 Eug 16828 o 62 Eug 4049 o 28 Eug 8215 o 63 Eug 40511 o 29 Eug 8220 o 64 Eug 4056 o 30 Eug 16842 o 65 Eug 40514 o 31 Eug 8756 o 66 Eug 4065 o 32 Eug 16871 o 67 Eug 4063 o 33 Eug 9218 o 68 Eug 4066 o 34 Eug 44211 o 69 Eug 0615 o 35 Eug 2242 o 70 Eug 40614 o

Table 1

(second pick)

Seventy strains were selected in the initial experiment, and repeated experiments were performed while selecting strains having high activity of inhibiting the attachment of Helicobacter pylori. Among these lactic acid bacteria, 35 bacterial strains capable of reducing the attachment number of Helicobacter pylori to about 10 ⁵ or less were selected, as shown in Table 2. Bacteria name The number of attached Helicobacter pylori (inhibiting attachment ability<10 ⁶ ) Eug 6014 o Eug 6017 o Eug 0340 o Eug 0352 o Eug 0442 o Eug 8126 o Eug 8130 o Eug 8540 o Eug 6713 o Eug 6123 o Eug 20553 o Eug 8215 o Eug 16842 o Eug 8756 o Eug 4416 o Eug 4419 o Eug 4427 o Eug 4439 o Eug 24013 o Eug 0618 o Eug 40312 o Eug 0317 o Eug 0315 o Eug 40410 o Eug 0412 o Eug 6404 o Eug 4047 o Eug 4048 o Eug 4049 o Eug 4056 o Eug 4065 o Eug 4063 o Eug 4066 o Eug 0615 o Eug 14439 o

Table 2

(third pick)

The 35 lactic acid bacteria obtained from the secondary selection were repeated again, and 22 strains were selected, which are shown in Table 3. Bacteria name The number of attached Helicobacter pylori Eug 6017 <10 ⁴ Eug 0352 2.7×10 ⁵ Eug 8126 4.4×10 ⁴ Eug 6713 1.4×10 ⁵ Eug 6123 2.4×10 ⁴ Eug 16842 8.3×10 ⁴ Eug 8756 1.4×10 ⁴ Eug 4416 <10 ⁴ Eug 4419 <10 ⁴ Eug 4427 7.8×10 ⁴ Eug 0618 9.1×10 ⁴ Eug 0317 2.8×10 ⁵ Eug 0315 3.5×10 ⁵ Eug 0412 3.5×10 ⁴ Eug 6404 <10 ⁴ Eug 4047 <10 ⁴ Eug 4048 <10 ⁴ Eug 4049 8.7×10 ⁴ Eug 4065 8.1×10 ⁵ Eug 4063 4.1×10 ⁴ Eug 4066 <10 ⁴ Eug 0615 2.1×10 ⁴

table 3

The identification of embodiment 3 bacterial strains

According to the BIOLOG kit and the Bergey manual, the results of identifying the 22 lactic acid bacteria strains selected in Examples 1 and 2 are shown in Table 4. Bacteria name Gram identification Cell morphology Spore form Catalase Air permeability (facultative bacteria) athleticism Eug 6017 (+) Streptococcus (-) (-) (+) (-) Eug 0352 (+) Streptococcus (-) (-) (+) (-) Eug 8126 (+) Streptococcus (-) (-) (+) (-) Eug 6713 (+) cocci (-) (-) (+) (-) Eug 6123 (+) Corynebacterium brevis (-) (-) (+) (-) Eug 16842 (+) Streptococcus (-) (-) (+) (-) Eug 8756 (+) Corynebacterium brevis (-) (-) (+) (-) Eug 4416 (+) Corynebacterium brevis (-) (-) (+) (-) Eug 4419 (+) Corynebacterium brevis (-) (-) (+) (-) Eug 0618 (+) cocci (-) (-) (+) (-) Eug 0317 (+) Streptococcus (-) (-) (+) (-) Eug 0315 (+) Streptococcus (-) (-) (+) (-) Eug 0412 (+) Streptococcus (-) (-) (+) (-) Eug 6404 (+) Streptococcus (-) (-) (+) (-) Eug 4047 (+) Streptococcus (-) (-) (+) (-) Eug 4048 (+) Corynebacterium longum (-) (-) (+) (-) Eug 4049 (+) Streptococcus (-) (-) (+) (-) Eug 4065 (+) Streptococcus (-) (-) (+) (-) Eug 4063 (+) Streptococcus (-) (-) (+) (-) Eug 4066 (+) cocci (-) (-) (+) (-) Eug 0615 (+) Streptococcus (-) (-) (+) (-)

Note: Streptobacillus, Coccobacillus, Short rod, Long rod

Table 4

As shown in Table 4, one strain of Leuconostococcus citrophilum was identified by the BIOLOG system, and 21 strains of Lactobacillus were identified by the method provided in the Bergey manual, all of which had the property of inhibiting the attachment of Helicobacter pylori to cells .

(four picks)

The 22 bacterial strains obtained by three times of selection were repeated for the third time, and 9 bacterial strains with high inhibitory activity were selected. The selection at this stage is not based on the order of inhibitory activity, but is divided and selected according to the shape of the bacteria. The selection results are shown in Table 5. Bacteria name The number of attached Helicobacter pylori Eug 6017 <10 ⁴ Eug 8126 1.2×10 ⁵ Eug 6713 1.4×10 ⁵ Eug 4419 <10 ⁴ Eug 4427 1.2×10 ⁵ Eug 6404 <10 ⁴ Eug 4047 <10 ⁴ Eug 4048 <10 ⁴ Eug 4066 <10 ⁴

table 5

(last pick)

The 9 lactic acid bacteria strains selected above were subjected to the fourth repeated test, one of each type of bacteria was selected, and finally 5 strains were selected, which are shown in Table 6. Bacteria name The number of attached Helicobacter pylori Eug 6017 <10 ⁴ Eug 4427 2.5×10 ⁵ Eug 6404 2.6×10 ⁴ Eug 4048 <10 ⁴ Eug 4066 <10 ⁴

Table 6

As shown in Table 6, among the 5 finally selected bacterial strains, the strain of Leuconostoc citrophila was named Leuconostoc citrophila EUG4427 (KCCM-10274), and the strains of the 4 Lactobacillus species were respectively Named Lactobacillus EUG6017 (KCCM-10273), Lactobacillus EUG4066 (KCCM-10270), Lactobacillus EUG4048 (KCCM-10271), and Lactobacillus EUG6404 (KCCM-10272). These 5 strains were deposited at the Korean Microbial Preservation Center on May 16, 2001 under the above names.

Example 4 Determination of Growth Inhibition of Helicobacter pylori by Kimchi Lactic Acid Bacteria Culture Broth

Leuconostoc citricophilus EUG4427 (KCCM-10274), Lactobacillus EUG6017 (KCCM-10273), Lactobacillus EUG4066 (KCCM-10270), Lactobacillus EUG4048 (KCCM-10271) and Lactobacillus selected from Example 3 EUG6404 (KCCM-10272) were cultured in MRS liquid medium at 37°C for 48 hours, and then centrifuged to obtain the supernatant.

A 25 ml agar plate supplemented with 10% equine serum Brucella agar medium was prepared and dried, and 0.5 ml of Helicobacter pylori suspension (1×10 ⁸ CFU/mL) was spread thereon and dried for 5 minutes.

The medium was punched with a sterilized cork cone, 100 μl of the centrifuged supernatant of each strain was introduced, and cultured at 37° _C for 72 hours under 10% CO2 partial pressure to measure the size of the transparent ring they formed. MRS liquid medium was used instead of the supernatant as a control. The growth inhibition results of lactic acid bacteria against Helicobacter pylori in vitro are shown in Table 7. strain name Radius of transparent ring (mm) control 0 Leuconostoc citricophilus EUG 4427 (KCCM-10274) 3.0 Lactobacillus EUG 6404 (KCCM-10272) 4.5 Lactobacillus EUG 4048 (KCCM-10271) 4.0 Lactobacillus EUG 6017 (KCCM-10273) 2.5 Lactobacillus EUG 4066 (KCCM-10270) 4.0

Table 7

As shown in Table 7, the culture result of 5 kinds of Kimchi lactic acid bacteria supernatants isolated from Example 3 shows that the radius of the transparent ring produced is between 2.5mm and 4.5mm, indicating that Kimchi lactic acid bacteria of the present invention The bacterium possesses outstanding activity inhibiting the growth of Helicobacter pylori.

Example 5 Inhibition of Helicobacter pylori attachment to gastric cancer cells

Human Kato III gastric cancer cells were cultured on RPMI1640 medium containing 10% FBS at 37°C and 10% CO ₂ . Helicobacter pylori was cultured on Brucella agar medium containing 10% horse serum at 37°C and 10% CO ₂ for 48 hours, and then the colonies were collected and suspended in saline. 0.5 ml of 1×10 ⁶ Kato III suspension and 0.25 ml of lactic acid bacteria culture solution were reacted at 37° C. for 30 minutes, and slowly shaken while reacting. The culture medium was replaced by MRS liquid medium as a control.

0.25 ml of Helicobacter pylori suspension (1×10 ⁸ CFU/mL) was added to the above reaction solution, and the reaction was continued for 90 minutes, then washed with saline three times, and centrifuged at 1000 rpm for 5 minutes each time. The number of H. pylori attached to Kato III was then determined by urease assay.

Urease breaks down urea to produce ammonia, which can be quantified using phenol/nitroprusside and alkaline hypochlorite. If urease exists, the reaction liquid turns from colorless to bright blue, and the activity of urease is determined by measuring the absorbance at 625 nm. The number of Helicobacter pylori is calculated based on urease activity. When the number of Helicobacter pylori is 10 ⁴ -10 ⁷ , the standard curve of this experiment shows a linear region.

The activities of the five Kimchi lactic acid bacteria obtained in Example 3 to inhibit the attachment of Helicobacter pylori to human gastric cancer cells are shown in Table 8. strain name The number of attached Helicobacter pylori (CFU) control >10 ⁷ Leuconostoc citricophilus EUG 4427 (KCCM-10274) 2.5×10 ⁵ Lactobacillus EUG 6404 (KCCM-10272) 2.6×10 ⁴ Lactobacillus EUG 4048 (KCCM-10271) <10 ⁴ Lactobacillus EUG 6017 (KCCM-10273) <10 ⁴ Lactobacillus EUG 4066 (KCCM-10270) <10 ⁴

Table 8

It can be confirmed from Table 8 that the 5 kinds of Kimchi lactic acid bacteria obtained in Example 3 only make a small amount of Helicobacter pylori attach to Kato III human gastric cancer cells, indicating that 5 kinds of lactic acid bacteria obtained from Kimchi of the present invention can inhibit Helicobacter pylori attached to the gastrointestinal tract.

The preparation of the high-function kimchi that embodiment 6 protects gastrointestinal tract

Leuconostoc citricophilus EUG4427 (KCCM-10274), Lactobacillus EUG6017 (KCCM-10273), Lactobacillus EUG4066 (KCCM-10270), Lactobacillus EUG4048 (KCCM-10271) and Lactobacillus EUG6404 ( KCCM-10272) was freeze-dried and made into powder, one or more bacterial strains were mixed, and then the bacteria containing 10 ⁶ -10 ¹¹ per kilogram of kimchi, preferably 10 ¹⁰ , were mixed with the kimchi seasoning, and then passed A conventional method prepares high-function kimchi that protects the gastrointestinal tract.

The kimchi lactic acid bacteria of the present invention can inhibit the Helicobacter pylori parasitic on the gastric mucosa tissue from attaching to the gastrointestinal tract and inhibit its growth, so it is very effective for killing the Helicobacter pylori and preventing reinfection. In addition, Kimchi containing Kimchi lactic acid bacteria capable of protecting the gastrointestinal tract is very effective for the prevention and treatment of gastritis, gastric ulcer, and duodenal ulcer caused by Helicobacter pylori.

In addition, lactic acid bacteria are not derived from the human body or milk fermentation products, but lactic acid bacteria with the required functions are derived from kimchi, which is the main source of lactic acid bacteria for Koreans. Therefore, the described function can be added to the traditional food kimchi and other foods, so that the supply source of functional lactic acid bacteria is more diversified. Applicant or Agent File Number OPP 020479KR International Application No. PCT/KR02/00899

                                                                                                                                                                                                                                                                  Explanation on the Preservation of Microorganisms

(Rule 13bis)

Form PCT/RO/134 (July 1992) Applicant or Agent File Number OPP 020479KR International Application No. PCT/KR02/00899

                                                                                                                                                                                                                                                                  Explanation on the Preservation of Microorganisms

(Rule 13bis)

Form PCT/RO/134 (July 1992) Applicant or Agent File Number OPP 020479KR International Application No. PCT/KR02/00899

                                                                                                                                    Explanation on the Preservation of Microorganisms

(Rule 13bis)

Form PCT/RO/134 (July 1992) Applicant or Agent File Number OPP 020479KR International Application No. PCT/KR02/00899

                                                                                                                                                                                                                                                                  Explanation on the Preservation of Microorganisms

(Rule 13bis)

Form PCT/RO/134 (July 1992) Petitioner or Agent File No. OPP 020479KR International Application No. PCT/KR02/00899

                                                                                                                                                                                                                                                                  Explanation on the Preservation of Microorganisms

(Rule 13bis)

Form PCT/RO/134 (July 1992)

Claims (7)

1. Inhibit the growth of Helicobacter pylori and inhibit its attachment to Kimchi lactic acid bacteria in the gastrointestinal tract. 2. The kimchi lactic acid bacteria according to claim 1, wherein the kimchi lactic acid bacteria are isolated from kimchi, which is ripened at 1 to 40° C. for 90 days or until the pH value reaches about 3.0 after the kimchi is prepared. . 3. The kimchi lactic acid bacterium according to claim 1, wherein the kimchi lactic acid bacterium belongs to Leuconostoc species or Lactobacillus species. 4. The kimchi lactic acid bacterium according to claim 1 or 2, wherein the kimchi lactic acid bacterium is one or more selected from the group consisting of Leuconostoc citricophilus EUG4427 (KCCM-10274) , Lactobacillus EUG6017 (KCCM-10273), Lactobacillus EUG4066 (KCCM-10270), Lactobacillus EUG4048 (KCCM-10271) and Lactobacillus EUG6404 (KCCM-10272). 5. A functional food useful for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori, comprising the kimchi lactic acid bacteria according to any one of claims 1 to 4. 6. A high-function gastrointestinal tract-protecting kimchi useful for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer, comprising the kimchi lactic acid bacteria according to any one of claims 1 to 4. 7. A medicament for the prevention and treatment of gastritis, gastric ulcer and duodenal ulcer caused by Helicobacter pylori, comprising the kimchi lactic acid bacteria as claimed in any one of claims 1 to 4.