KR101737332B1 - New Enterococcus faecium L11 and probiotics composition comprising the same - Google Patents

Abstract

The present invention relates to a novel Enterococcus faecium L11 strain and a prodrug composition containing the same as an active ingredient. The Enterococcus faecium L11 strain or its culture according to the present invention has antimicrobial effect against various pathogenic microorganisms and is excellent in acid resistance, artificial gastric tolerance, biliary properties and immunity enhancing effect, so that it can be used as a probiotic agent, a medicament, a health food, , Feed additives, and the like.

Description

[0001] The present invention relates to a novel Enterococcus faecium L11 strain and a probiotic composition comprising the same as an active ingredient.

The present invention relates to a novel Enterococcus faecium L11 strain and a prodrug composition containing the same as an active ingredient.

Lactic acid bacteria are widely distributed in various kinds of fermented foods, minerals of people and animals, and soil, and are closely related to our lives. Lactic acid bacteria maintains a balance of intestinal microorganisms and produces bacteriocin, a large amount of lactic acid and acetic acid to inhibit the growth of food spoilage bacteria, (Ahn SB et al., J. Biomed. Res. 13 (4): 353-356, 2012).

Therefore, in recent years, studies have been actively conducted to use various lactic acid bacteria as probiotics. Probiotic agents are responsible for the microorganisms that have beneficial effects on host health and feed efficiency by producing a variety of metabolites useful for the host, with antimicrobial activity and enzyme activity, which helps the intestinal microbial balance (fuller, R. J appl bacteriol. 66 (5): 365-378, 1989), and are used in various fields such as formulases, lactic acid bacteria, feed additives, and health foods. In general, to be used as a probiotic, it is necessary to ingest orally to reach the colon and rectum, to take the intestinal tract of the host as a habitat, and to be non-pathogenic and non-toxic. Therefore, the probiotic agent should basically have resistance to the intestinal environment (acid, enzyme, bile) and excellent functionality (antimicrobial agent production ability, immunity promoting ability, inhibiting ability of pathogenic bacteria, fixation property) It is true.

On the other hand, microorganisms in Enterococcus have been studied as raw material microorganisms for the production of probiotics. Prior arts include Korean Patent No. 10-0468522 and Korean Patent No. 10-0523255.

Accordingly, the inventors of the present invention have focused on the fact that a strain of Enterococcus can be used as a probiotic, and as a result, they have identified a new Enterococcus pneumatosis strain, and found that the strain and its culture have acid resistance, The present inventors have completed the present invention by discovering that there is an antibacterial activity and an immunity enhancing effect on microorganisms.

A problem to be solved by the present invention is to provide a novel strain Enterococcus cephem L11 strain of Accession No. KFCC11572P, which is a novel strain, and is also useful as a prodrug or a pathogenic microorganism containing the strain or a culture thereof as an active ingredient A pharmaceutical composition for preventing or treating a disease caused by pathogenic microorganisms, a food composition for preventing or ameliorating a disease caused by a pathogenic microorganism, a lactic acid bacterial preparation, a fermented milk and a feed additive.

In order to solve the above problems, the present invention provides Enterococcus faecium L11 strain of Accession No. KFCC11572P. At this time, the strain may be isolated from chicken.

In addition, the strain may induce the production of cytokines IL-6 and TNF-a.

Further, the present invention provides a biocidal composition comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a pharmaceutical composition for preventing or treating a disease caused by a pathogenic microorganism comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a food composition for preventing or ameliorating a disease caused by a pathogenic microorganism comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a lactic acid bacterial preparation comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a fermented milk comprising the strain or a culture thereof as an active ingredient.

The present invention also provides a feed additive comprising the strain or a culture thereof as an active ingredient.

The Enterococcus cephem L11 strain or its culture of Accession No. KFCC11572P according to the present invention has antibacterial effect against various pathogenic microorganisms and is excellent in acid resistance, tolerance to artificial gastric juice, bile resistance and immunity enhancement, Food, lactic acid bacterial preparation, feed additive and the like.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a phylogenetic location of the Enterococcus faecium L11 strain according to the present invention.
FIG. 2 shows the antimicrobial activity of Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199 in the culture medium of the present invention. FIG. 2a shows the antimicrobial activity of Vancomycin-resistant Enterococcus faecalis And FIG. 2B is a photograph showing the antimicrobial activity against Enterococcus faecalis ATCC 19433, wherein the left side shows the pH before the adjustment and the right side shows the pH after 7.0.
FIG. 3 is a graph showing the result of measuring the bow performance of the Enterococcus faecium L11 strain treated with macrophages according to the present invention.
FIG. 4 is a graph showing the results of measuring the concentration of cytokine IL-6 by ELISA after treatment of Enterococcus faecium L11 strain according to the present invention with macrophages.
FIG. 5 is a graph showing the results of measuring the concentration of cytokine TNF- by ELISA after treating the Enterococcus faecium L11 strain of the present invention according to the present invention with macrophages.

Hereinafter, the present invention will be described in more detail.

The present invention provides a novel Enterococcus faecium L11 strain of Accession No. KFCC11572P.

The strains were prepared by isolating various lactic acid bacterial species from the cecum of healthy chickens and analyzing the physiochemical characteristics of the isolated strains to identify strains having excellent antimicrobial activity and immunity enhancing effect on acid resistance, bile acid resistance and harmful microorganisms. The strain was designated Enterococcus faecium L11 and deposited on Dec. 4, 2013 at the Korean Microorganism Conservation Center to receive the deposit number KFCC11572P.

The strain of the present invention has antibacterial activity against harmful microorganisms. In particular, it has high antimicrobial activity against antibiotic resistant strains such as Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 33592, Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199, Extended spectrum β-lactamase (ESBL) producing E. coli, It also has a high antimicrobial activity against foodborne pathogens such as typhimurium ATCC 13311, Salmonella enteritidis ATCC 13076, and Escherichia coli O157: H7 ATCC 35150. In addition, the strain of the present invention is excellent in acid resistance and bile acid resistance, and is particularly effective in inducing the secretion of cytokines, particularly the production of cytokines IL-6 and TNF-a, And can play an important role in regulating the immune system of the human body.

Further, the present invention provides a biocidal composition comprising the strain or a culture thereof as an active ingredient. At this time, the culture may include both the concentrated liquid and the dried material of the culture liquid.

The Enterococcus faecium L11 strain or the culture thereof according to the present invention is excellent in antimicrobial activity, acid resistance, bile resistance and immunity enhancing effect against harmful microorganisms. Therefore, the Enterococcus cephem L11 strain or the culture thereof of the present invention can be effectively used for a prophylactic agent, particularly a formulant, a lactic acid bacterial preparation, a feed additive, and a health food, which are human medicines.

The present invention also provides a composition for preventing, treating or ameliorating a disease caused by a pathogenic microorganism comprising the strain or a culture thereof as an active ingredient.

The composition comprises a pharmaceutical composition or a food composition. The pathogenic microorganism is preferably a pathogenic microorganism having antibiotic resistance or causing food poisoning, and the pathogenic microorganism causing the food poisoning is preferably Salmonella, Escherichia coli, Bacillus cereus strain, but is not limited thereto.

The disease is preferably selected from the group consisting of acute diarrhea, enteritis, gastroenteritis, constipation, abdominal pain, abdominal distension, cholera and food poisoning, but is not limited thereto.

The composition of the present invention can be administered orally or parenterally at the time of clinical administration and can be administered to an effective amount of the Enterococcus faecium L11 strain or a culture thereof as a main component, or a pharmaceutically acceptable carrier or one or more Two or more kinds of additives may be selected to prepare a composition of a conventional pharmaceutical formulation.

The carrier may be selected from one or more selected from among diluents, lubricants, binders, disintegrants, sweeteners, stabilizers and preservatives. Examples of the additives include one or more of flavorings, vitamins and antioxidants Can be used.

Examples of the diluent include lactose, corn starch, microcrystalline cellulose, and mannitol. The lubricant is preferably magnesium stearate or talc, and the binder The zero is preferably selected from polyvinylpyrrolidone or hydroxypropylcellulose. The disintegrant is preferably selected from carboxymethylcellulose calcium, sodium starch glycolate, polacrilin potassium or crospovidone, and the sweetener is selected from white sugar, fructose, sorbitol or aspartame. Sodium carboxymethylcellulose , Beta-cyclodextrin or xanthan gum, and the preservative is preferably selected from methyl parahydroxybenzoate, propyl p-hydroxybenzoate or potassium sorbate.

The composition of the present invention is used for the purpose of effectively preventing or treating a disease caused by a pathogenic microorganism, and is not particularly limited in its formulation. Examples of the composition include beverage, powder, capsule, soft capsule, tablet, gum, And can be used as a transdermal dosage form such as a lotion, ointment, gel, cream, patch, or spray, and can be used as a tablet, a pill, a tablet, a powder or a granule.

In the composition of the present invention, a minerals-containing compound such as vitamin B, C, or E, beta-carotene, Ca, or Mg may be used as an auxiliary component, or a phospholipid such as lecithin or a compound such as maltol or oligosaccharide may be used.

In addition to the above components, known additives include natural flavors such as plum, lemon flavor, pineapple flavor or herb flavor, natural flavors such as natural fruit juice and flavonoid, sweeteners such as honey, honey, sugar and citric acid, sodium citrate The same acid agent can be mixed and used.

In addition, the health food containing the strain or the culture of the present invention can be processed into various forms such as tea, jelly, juice, extract, beverage and the like.

When the strain of the present invention or a culture thereof is used as a food additive, each of the strains or a culture thereof may be directly added or used together with other food or food ingredients.

Examples of the food to which the strain of the present invention or the culture thereof can be added include meat, sausage, bread, candy, snacks, confectionery, dairy products, various soups, drinks, tea, Drink, and the like, and includes all conventional health foods.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of natural carbohydrates include glucose, fructose, maltose, dextrin and xylitol. Natural sweeteners such as tau martin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is 0.01 to 0.04 g, preferably 0.02 to 0.03 g per 100 ml of the strain or the culture of the present invention.

The strain or culture thereof may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, organic acids, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonating agents and the like.

The present invention also provides a lactic acid bacterial preparation comprising the strain or a culture thereof as an active ingredient.

The strain or culture thereof of the present invention is preferably 0.01 to 50 parts by weight per 100 parts by weight of the lactic acid bacterial preparation, but is not limited thereto and can be appropriately adjusted by a person skilled in the art.

The present invention also provides a fermented milk comprising the strain or a culture thereof as an active ingredient.

The strain or the culture thereof of the present invention is preferably 0.01 to 50 parts by weight per 100 parts by weight of the fermented milk, but is not limited thereto and may be appropriately adjusted by a person skilled in the art.

The present invention also provides a feed additive comprising the strain or a culture thereof as an active ingredient.

The strain or culture of the present invention is preferably 0.01 to 50 parts by weight per 100 parts by weight of the feed additive, but is not limited thereto and can be appropriately adjusted by a person skilled in the art.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments and the like. It will be apparent to those skilled in the art, however, that these examples are provided for further illustrating the present invention and that the scope of the present invention is not limited thereto.

Example  1: Isolation and selection of strains

In order to isolate strains from the cecum of healthy chickens, 1 g of the contents was added to 10 g of sterilized water, and 1 ml of the diluted solution was taken and diluted stepwise at 10 ⁵ , 10 ⁶ , and 10 ⁷ magnification. Each 0.1 ml of each dilution was blotted onto MRS (Difco, USA) agar medium (MRS, De Man, Rogosa and Sharpe) and cultured at 37 ° C for 48 hours. Colony was collected from each medium and 20 strains Were selected. In order to confirm the inhibitory effect against the harmful microorganisms of the first selected strain, the spot-on-lawn method was applied to Salmonella enteritidis ATCC 13076, Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 33592, Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199 After the experiment, the strain with the best inhibitory effect was selected.

Example  2: Identification of isolated and sorted strains

The morphological, physiological and biochemical characteristics of the selected strains were examined and the strains were finally identified by analyzing 16S rDNA gene sequences. The physiological and biochemical characteristics were examined after 48 hours of incubation at 37 ° C using API 50 CHL kit (Biomerieux, France). The results are shown in Table 1 below . The + in Table 1 means a positive reaction, and the negative sign means a negative reaction.

Characteristics Reaction Characteristics Reaction Morphology shape cocci Arbutin + Gram stain + Esculin + Control - Salicin + Glycerol - Celiobiose + Erythritol - Maltose + D-arabinose - Lactose + L-arabinose + Melibiose - Ribose D-ribose + Sucrose + D-xylose - Trehalose + L-xylose - Inulin - D-adonitol - Melezitose - Methyl-BD-xylopyranosicle - Raffinose - D-galactose + Starch - D-glucose + Glycogen - D-fructose + Xylitol - D-mannose + Gentiobiose + L-sorbose - D-turanose - Rhamnose - D-lyxose - Dulcitol - D-tagatose + Inositol - D-fucose - Mannitol + L-fucose - Sorbitol - D-arabitol - alpha -Methyl-D-mannoside - L-arabitol - alpha -Methyl-D-glucoside - Gluconatee - N-Acethyl-glucosamine + 2-keto-Gluconate - Amygdalin + 5-keto-gluconate -

The 16S rDNA gene sequence (Sequence Listing 1) was analyzed and the morphological, physiological, biochemical and genetic characteristics of the isolated strains were investigated. As a result, it was confirmed that the strain had high homology with Enterococcus faecium. Therefore, the strain of the present invention was identified as an Enterococcus faecium strain and designated as Enterococcus faecium L11, deposited at the Korean Microorganism Conservation Center on Dec. 4, 2013 and deposited under accession number KFCC11572P .

Example  3: Acid resistance  And artificial gastric tolerance test

The acid tolerance was determined by diluting the strain Enterococcus lethal L11 according to the present invention in a MRS liquid medium at 37 ° C. for 48 hours and then diluting it to a fresh liquid medium adjusted to pH 2 and 3 with 3M hydrochloric acid to 1 × 10 ⁷ CFU / Respectively. The diluted cells were incubated at 37 ° C for 3 hours. After incubation, the cells were continuously diluted with sodium phosphate buffer (0.1 M, pH 6.2) to neutralize the acidity of the medium, and the number of viable cells was measured on the MRS plate medium after culturing at 37 ° C for 48 hours. The survival rate was calculated by comparing the initial bacterial concentration with the ratio of colonies of bacteria formed on the MRS plate medium. The results are shown in Table 2 below.

The measurement of artificial gastric tolerance was carried out by sterilizing the MRS liquid medium adjusted to pH 3 with 3M hydrochloric acid, and then adding 1% pepsin to measure artificial gastric juice. The experiment was then carried out in the same manner as the acid resistance measurement described above And the results are shown in Table 2 below.

Condition Survival rate (%) after 3 hours  pH 2 64.1 ± 2.2  pH 3 70.2 ± 1.5  pH 3 + pepsin 69.9 ± 1.3

In order for the strain to function as a probiotic, it must pass through a stomach with a pH of 2.5 to 3.5 (Holzapfel, WH et al., International Journal of Food Microbiology 41: 85101, 1998) It should show high survival rate over time. The acidity of the probiotic can be regarded as a criterion based on the survival rate after 2 hours at pH 3 (Dora IA Pereira and Glenn R. Gibson, Applied and Environmental Microbiology, 68: 9, 46894693, 2002) , And the survival rate of the strain Enterococcus cepum L11 according to the present invention after 3 hours at pH 3 is 70% or more.

Example  4 : My bile  inspection

The measurement of the biliary properties was carried out as follows. The Enterococcus faecium L11 strain according to the present invention was cultured in an MRS liquid medium at 37 ° C for 48 hours and then plated on MRS plate medium containing different concentrations (0.3, 1%) of Oxgall (Sigma, USA) The growth of the bacteria was observed for 48 hours.

Gilliland et al. (Journal of Dairy Science, 67 (12): 3045-3051, 1984) reported that tolerance to the bile solution to be fed by live bacteria having viable cell viability can be grown in a medium containing 0.3% oxgall, It should be able to grow in medium containing higher oxgall. As shown in the following Table 3, the strain Enterococcus cepum L11 according to the present invention was hardly affected by the medium containing 0.3% oxgall, and the survival rate of the initial number of bacteria was 99.0% in the medium containing 1% oxgall It can be confirmed that it has very strong biliary properties.

bile acid Survival rate (%) 0.3% 99.7 ± 0.1 One % 99.0 ± 0.7

Example  5: Antimicrobial activity measurement

The antimicrobial activity against pathogens was measured according to the disk diffusion method (NCCLS M2-A7, 2000) of the Clinical and Laboratory Standards Institute (CLSI; formerly NCCLS). 100 ml of MRS liquid medium was inoculated with 1% L11 pre-culture solution and cultured at 37 ° C for 48 hours. The culture was centrifuged at 8,000 g, and the supernatant was filtered with a 0.2 μm membrane filter to remove the cells. The supernatant was concentrated 10 times by using a freeze dryer, and the concentrated culture was filtered with a 0.2 μm membrane filter and used as a sample for measuring the antibacterial activity. The inoculum was diluted in sterile physiological saline solution to a concentration of 1.5 × 10 ⁸ CFU / ml on the Mueller Hinton agar medium and uniformly applied to the surface of the solid medium using a cotton swab. Then, 80 쨉 l of the antimicrobial activity measuring sample was placed in a culture medium and incubated in a thermostat at 37 ° C for 18-24 hours. The diameter (mm) of inhibition zone (growth inhibition zone) indicating growth inhibition was measured and shown in Table 4 below.

FIG. 2 shows the antimicrobial activity of the lactic acid bacteria culture solution according to the present invention. FIG. 2a shows antibacterial activity against Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199, 7.0, and FIG. 2B is a photograph showing the antibacterial activity against Enterococcus faecalis ATCC 19433, wherein the left side shows the pH before the adjustment and the right side shows the pH after 7.0.

As a result of the measurement, the antimicrobial substance secreted by the Enterococcus lepromus strain L11 according to the present invention was found to be resistant to methicillin-resistant Staphylococcus aureus (MRSA) ATCC 33592, Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199, The antimicrobial activity of antibiotic-resistant strains such as E. coli was high, and antimicrobial activity was also observed in foodborne pathogens such as Salmonella typhimurium ATCC 13311, Salmonella enteritidis ATCC 13076 and Escherichia coli O157: H7 ATCC 35150.

In order to confirm that the antimicrobial activity of the Enterococcus faecium L11 strain according to the present invention is due to the production of an antimicrobial substance instead of only the acid, the following experiment was conducted. 100 ml of the MRS liquid medium was inoculated with 1% of the pre-culture of Enterococcus faecium L11 and cultured at 37 ° C for 48 hours. The culture broth was centrifuged at 8,000 g, and the supernatant obtained was filtered with a 0.2 μm membrane filter, Respectively. The pH of the culture medium was adjusted to 7.0 to eliminate the antimicrobial activity due to the low pH of the culture medium, and then the concentration of the culture medium was adjusted to 10 times by using a freeze dryer. The concentrated culture was filtered with 0.2 μm membrane filter and used for the antibacterial activity test. The antimicrobial activity test is the same as the above, and the diameter (mm) of the inhibition zone (growth inhibition zone) indicating growth inhibition is observed and shown in Table 4 below.

As a result, it was possible to observe a high level of proliferation inhibition in Enterococcus such as Vancomycin-resistant Enterococcus faecalis (VRE) ATCC 52199, Enterococcus faecalis ATCC 19433, Enterococcus hirae ATCC 8043, Therefore, the Enterococcus faecium L11 strain according to the present invention was found to produce other antimicrobial substances in addition to the acid.

Test strain Inhibition zone (mm) pH 4 pH 7 Gram - positive bacteria Methicillin-resistant S. aureus (MRSA) ATCC 33592 13.8 ± 0.25 0 Staphylococcus aureus ATCC 29213 13.3 ± 0.25 0 Vancomycin-resistant E. faecalis (VRE) ATCC 52199 20.5 ± 0.5 16.5 ± 0.5 Enterococcus faecalis ATCC 19433 19.8 ± 0.75 17.3 ± 0.75 Enterococcus durans ATCC 6056 21 ± 0.5 20.5 ± 0.75 Enterococcus faecium 16.5 ± 0.25 15 ± 1.0 Enterococcus hirae ATCC 8043 22.5 ± 1.5 20 ± 0.5 Enterococcus galliranun JCM 8728 20 ± 0.75 14.3 ± 1.75   Bacillus cereus 12.5 ± 0.5 0 Gram - negative bacteria Extended spectrum-lactamase (ESBL) producing E. coli 14.3 ± 0.25 0 Escherichia coli ATCC 25922 15.8 ± 0.25 0 Escherichia coli O157: H7 ATCC 35150 13.5 ± 0.25 0 Salmonella enteritidis ATCC 13076 15.5 ± 1.0 0 Salmonella typhimurium ATCC 13311 14.5 ± 0.5 0 Pseudomonas aeruginosa ATCC 27853 20.0 ± 0.5 0

Example  6: Measurement of immunological activity

Immunoreactivity was measured by macrophage activity and production of cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) secreted from the macrophages. Experiments were conducted using the same method.

Test organisms were obtained as follows. 100 ml of MRS liquid culture medium was inoculated with 1% of the culture solution of Enterococcus faecium L11 according to the present invention and cultured at 37 DEG C for 48 hours. The culture broth was centrifuged at 8,000 xg and the cells were suspended in PBS (sodium phosphate buffered saline, pH 7.4). After recovering only the cells, they were heated at 121 ° C for 15 minutes, centrifuged at 8,000 × g for 10 minutes, and diluted with PBS.

Macrophage is a typical immune cell. It is an important primary immune cell that plays a role of phagocytosis of viruses or foreign substances penetrating the body and stimulates other immune organs in the body by generating various action factors . To measure macrophage activity, 2 ml of 3% thioglycollate medium (TG medium) was injected intraperitoneally in ICR (Imprinting Control Region) mice, and the induced macrophages were recovered within 48 hours. Then, RPMI (Roswell Park Memorial Institute) 1640 medium. Cells were adjusted to 110 ⁶ cells / mL, and 200 μL aliquots were added to 96-well plates. This culture (37 ℃, 2 hours) to form a macrophage monolayer was removed, and then the supernatant was dispensed to 180 μL of RPMI 1640 medium to each well and the addition of L11 dead cells Sample 20 μL 37 ℃ in 5% CO ₂ incubator, And cultured in a 5% CO ₂ incubator for 1 day. After incubation, we measured the activity of macrophages by performing the assay of 2- (4-iodophenyl) -3- (4-nitrophenyl) -5- (2,4-disulfophenyl) -2H-tetrazolium monosodium salt reagent Respectively. 10 μg / ml of lipopolysaccharide (LPS, Sigma Co.) was used as a positive control sample and PBS was used as a negative control sample (FIG. 3) to evaluate the activity of the sample. Enterococcus faecium L11 had a higher activity than the negative control (PBS), and the highest concentration (1 × 10 ⁸ cfu / ml) of the enzyme was 67%.

IL-6, IL-8 (CXCL8), IL-12 and TNF-α are the anti-inflammatory cytokines that are produced in macrophages. IL-6 and TNF-α play a key role in inducing an acute phase reaction, which effectively causes host defense by various methods (Peter, Life science publishing Co., 233-246. 2006) . Renshaw et al. (The Journal of Immunology 169 (9): 4697-4701, 2002) showed that a decrease in secretion of IL-6 and TNF-α secreted from macrophages indicates a marked decrease in immune function, In the study, cytokine IL-6 and TNF-α were measured. The amount of IL-6 and TNF-a produced and secreted in macrophages was measured using mouse macrophages and L11 germ samples derived from 3% TG medium. IL-6 and TNF-α ELISA kit (BD Opt EIA, BD Biosciences, USA) were used for cytokine assay. The results of the cytokine IL-6 test are shown in Fig. 4, and the test results of TNF-a are shown in Fig.

4 and 5, enterococcus leposterol L11 according to the present invention was found to induce cytokine IL-6 and TNF-? Compared to the negative control (PBS), and in the case of TNF-? Cytokine production was higher than that of control (LPS).

Korea Microorganism Conservation Center (Domestic) KFCC11572P 20131204

Claims (10)

Enterococcus faecium L11 strain of Accession No. KFCC11572P. The method according to claim 1,
Wherein said strain is isolated from chicken. ≪ RTI ID = 0.0 > 11. < / RTI > The method according to claim 1,
Wherein said strain induces the production of cytokines IL-6 and TNF- [alpha]. A prophylactic composition comprising the Enterococcus faecium L11 strain of claim 1 or a culture thereof as an active ingredient. By one or more pathogenic microorganisms selected from the group consisting of acute diarrhea, enteritis, gastroenteritis, constipation, abdominal pain, abdominal distension, cholera and food poisoning comprising the Enterococcus faecium L11 strain of claim 1 or a culture thereof as an active ingredient A pharmaceutical composition for preventing or treating a disease that occurs. delete By one or more pathogenic microorganisms selected from the group consisting of acute diarrhea, enteritis, gastroenteritis, constipation, abdominal pain, abdominal distension, cholera and food poisoning comprising the Enterococcus faecium L11 strain of claim 1 or a culture thereof as an active ingredient A food composition for preventing or ameliorating a disease that occurs. A lactic acid bacterial preparation comprising the Enterococcus faecium L11 strain of claim 1 or a culture thereof as an active ingredient. A fermented milk comprising the Enterococcus lepthae L11 strain of claim 1 or a culture thereof as an active ingredient. A feed additive comprising the Enterococcus faecium L11 strain of claim 1 or a culture thereof as an active ingredient.

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