KR101487446B1 - Lactobacillus mucosae LM1 with excellent acid-Resistant, bile-resistance, antibacterialactivity and adhere to mucosal surfaces, and composition containing the same - Google Patents

Abstract

The present invention relates to Lactobacillus mucosae LM1 having excellent acid resistance, bile resistance, harmful bacteria controlling functions, and mucosal surface adsorbing functions and to a composition including the same. More specifically, the Lactobacillus mucosae LM1 is a Lactobacillus used for probiotics. Compared to existing Lactobacilli, the Lactobacillus mucosae LM1 has excellent mucosal surface adsorbing functions along with acid resistance, bile resistance, and harmful bacteria controlling functions. According to the present invention, general Lactobacilli have acid resistance, bile resistance, and harmful bacteria controlling functions. However, the Lactobacillus mucosae LM1 according to the present invention not only has the basic properties of the general Lactobacilli but also has excellent mucosal surface adsorbing functions so as to be usefully used for probiotics. The Lactobacillus mucosae LM1 has a high survivability coefficient on the mucosal surface so that intestinal regulation can be effectively performed in the intestines.

Description

[TECHNICAL FIELD] The present invention relates to a lactobacillus mucosa LM1 lactic acid bacterium having excellent acid resistance, brittleness, harmful bacteria inhibiting ability and intestinal adsorption ability, and a composition containing the lactic acid bacteria. composition containing the same}

The present invention relates to Lactobacillus mucosse LM1 ( Lactobacillus sp.), Which is excellent in acid resistance, biliary properties, mucosae More particularly, the present invention relates to Lactobacillus < RTI ID = 0.0 > Mucosal LM1 < / RTI > which is superior in absorption ability to intestinal acid, dyspepsia, ≪ / RTI >

In the human intestines, about 1 kg of microorganisms are inhabited, and the amount of food and the amount of microorganisms are almost the same, and the contents of the feces to be excreted every day occupy about 40% of bacteria except moisture. Microscopic examination of human feces reveals that it is composed of almost homogeneous masses. About 99% of these fungi are anaerobes. In healthy infants who are breastfed, more than 90% of the fecal bacterium is composed of Bifidobacterium , but as age increases, Bifidobacterium is decreased and intestinal bacteria are increased. It is a function of probiotics to help maintain the distribution of intestinal microflora in a healthy state during this normal aging process. Probiotics are strains that are beneficial to the intestinal environment when ingested and reached the intestines. That is, probiotics that reach the intestines and grow in the intestinal mucosa produce lactic acid, which makes the intestinal environment acidic. The number of harmful bacteria that can not survive in the acidic environment is reduced, and beneficial bacteria that grow well in acidic environment are further proliferated, thereby making the intestinal environment healthy.

Most probiotics known to date are lactic acid bacteria and some Bacillus And the like. Russian scientist Elie Mechinikoff has been studying the functioning of lactobacilli and probiotics since the Nobel Prize for discovering that Bulgarians enjoy longevity because of the ingestion of fermented milk fermented with Lactobacillus . In order to be recognized as probiotics, bacteria such as lactic acid bacteria should be able to survive in stomach acid and bile acid to reach the small intestine and proliferate and settle in the intestines and have the ability to adsorb intestinal epithelial cells as well as harmful bacteria .

There have been many reports of lactic acid bacteria having excellent acid resistance and biliary properties or having antimicrobial activity. However, there have been no studies on strains having the property of excellent intestinal cell adsorbability including the above characteristics.

Therefore, in the present invention, lactobacillus < RTI ID = 0.0 > LM1 < / RTI > ( Latobacillus < RTI ID = mucosae LM1) and to develop a composition comprising the lactic acid bacterium and the cultured product of the lactic acid bacterium, so as to be industrially applicable.

Novel Lactobacillus flutarium isolated from kimchi inhibiting the growth of pathogenic bacteria and its use

The object of the present invention is to separate Lactobacillus mucosae LM1 lactic acid bacterium from Lactobacillus mucosae LM1, which has excellent acid-fastness, anti-juice resistance and antibacterial activity as well as excellent adsorption capacity of the intestines, from the small intestine and colon of porcine probiotic lactic acid bacteria, A composition containing water, a composition for use in probiotics, and the like.

In order to achieve the above object, the present invention provides Lactobacillus mucosse LM1 ( Lactobacillus Lactobacillus Lactobacillus, Lactobacillus sp. mucosae LM1) KCCM11442P.

The present invention also relates to Lactobacillus mucosse LM1 ( Lactobacillus sp.), Which is excellent in acid resistance, mucosae LM1) KCCM11442P culture.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > mucosae LM1) KCCM11442P or a culture thereof.

The present invention also provides an antimicrobial composition characterized in that the antimicrobial composition is for antibacterial use against silver coli, Salmonella, Pseudomonas or Bacillus pathogens.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > mucosae LM1) KCCM11442P or a culture thereof.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > mucosae LM1) KCCM11442P or a culture thereof.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > mucosae LM1) KCCM11442P or a culture thereof.

The present invention also relates to the use of Lactobacillus < RTI ID = 0.0 > mucosae LM1) KCCM11442P or a culture thereof.

As described above, the lactobacillus mucosa LM1 lactic acid bacterium and the composition containing the same according to the present invention, which are excellent in acid resistance, brittle ability, harmful bacteria-suppressing ability and intestinal tract adsorption ability, are excellent in basic acid resistance for exhibiting the efficacy as probiotics, Lactobacillus mucosa LM1 having excellent intestinal adsorptivity as well as ability to inhibit bacteria and harmful bacteria can be identified and thereby a lactic acid bacterium capable of effectively acting in the intestines with a higher survival rate in the intestinal tract than conventional lactic acid bacteria and a composition containing the same Can be provided.

Figure 1. Scanning Electron Microscope of LM1 strain
Figure 2. 16s rDNA gene sequence of LM1 strain
Figure 3. Lactobacillus mucosa LM1 ( Lactobacillus acidity of mucosae LM1)
Figure 4. Lactobacillus mucosal LM1 endotoxicity studies
Figure 5. Comparison of mucin adsorption ability with Lactobacillus mucosa LM1 and other strains
Figure 6. Coagulation activity of Salmonella with Lactobacillus mucosa LM1 and other strains
7. Comparison of cell surface hydrophobicity with Lactobacillus mucosa LM1 and other strains
Figure 8. E. coli inhibition of Lactobacillus Lactobacillus LM1 culture concentrate
ASP: ammonium sulfate precititate

Hereinafter, the present invention will be described in more detail by way of specific examples. However, it is to be understood that the present invention is not limited by the following examples, and that various changes and modifications may be made therein without departing from the spirit and scope of the present invention.

Example  1. Isolation and Identification of Lactic Acid Bacteria

1-1. LM1  Isolation of lactic acid bacteria

In order for the lactic acid bacteria to settle well in the intestines, it is most important to combine them with the mucin surrounding the intestinal wall. Therefore, we tried to select the best bacteria among lactic acid bacteria that bind strongly to mucin.

First, 100 μl of pig-derived mucin (Sigma-Aldrich, USA) was coated on a 96-well polystyrene plate and fixed at 4 ° C. for 12 to 16 hours. Unfixed mucin was removed by washing three times with 10 mM Hepes-Hanks (HH) buffer solution. The small intestine and large intestine of the pigs were harvested and the intestinal contents were recovered, diluted with phosphate buffer (pH 7.0), and injected 200 μl into a mucin-coated 96-well plate (plate). After incubation at 37 ° C for 1 hour, the wells were washed with HH buffer to remove microorganisms that were not adsorbed to mucin or adsorbed weakly. Thereafter, the coated mucin layer was vigorously stirred to recover adsorbed microorganisms, which were diluted in stages and spread on MRS agar plates (Difico, USA). After culturing at 37 ° C for 24 hours, colonies cultured in the medium of the highest dilution were selected. Lactobacillus mucosae LM1 was identified by lactic acid bacteria selected by the sequencing method in Example 2 below . The characteristics of the Lactobacillus mucosae LM1 strain were similar to those of a typical Lactobacillus sp. Strain as a rod-shaped bacterium as shown in the electron microscope (Fig. 1).

1-2. LM1  16S of strain rDNA  Gene analysis

In order to identify the strain selected in Example 1-1, the chromosome of the selected strain was isolated and the 16S rDNA gene was amplified by PCR (Polymerase Chain Reaction) method and recovered. Then, the nucleotide sequence of the recovered 16S rDNA gene was analyzed, and the result is shown in FIG. Since the sequence analyzed using the BLAST (Basic Local Alignment Search Tool) search program of NCBI showed 99% homology with the 16S rDNA sequence of Lactobacillus mucosae , In the invention, the isolated strain was deposited under the Budapest Treaty on August 9, 2013 with the deposit number KCCM 11442P at the Korean Agency for Microbiological Research, the international depository institution.

1-3. Fermentation Characteristics of Selected Lactic Acid Bacteria

The fermentation characteristics of the sugar fermentation were investigated using API 50CHL kit (Bio Merioux, France) according to the supplier's experimental method. The sugar fermentation characteristics of the bacteria isolated in Example 1-1 are as shown in Table 1. The fermentation characteristics of the isolated strains were also identified as Lactobacillus mucosae , and the results of the identification of the 16S rDNA gene could be reaffirmed.

The sugar fermentation characteristics of the lactic acid bacteria selected in the present invention Party Decomposition Party Decomposition Glycerol - Esculin + Erythritol - Salicin - D-Arabinose - Cellobiose - L-arabinose + Maltose + Ribose + Lactose + D-Xylose + Melibiose + L-Xylose - Saccharose + Adonitol - Trehalose - β-Methyl-Xylose + Inulin - Galactose + Melezitose - D-Glucose + D-Raffinose + D-Fructose - Amidon - D-Mannose - Glycogen - L-Sorbose - Xylitol - Rhamnose - β-Genitobiose - Dulcitol - D-Lyxose - Inositol - D-Tagatose - Mannitol - D-Fucose - Sorbitol - L-Fucose - alpha -Methyl-D-Mannoside - D-arabitol - alpha -Methyl-D-Glucoside - L-Arabitol - N-acetyl-glucosamine - Gluconate + Amygdalin + 2-Keto-Gluconate + Arbutin - 5-Keto-Gluconate +

Example  2. Lactobacillus Mukose LM1 ( Lactobacillus mucosae LM1 )of Acid resistance  And My bile  Research

2-1. Acid resistance  Research

In order for lactobacilli to exert their potency as a probiotic in the intestine, they must pass over low pH after ingestion. In order to investigate the acid resistance of Lactobacillus mucosa LM1, the cells were inoculated in MRS broth medium (Difico, USA) and cultured at 37 ° C for 16-24 hours. Then, MRS liquid medium adjusted to pH 3.0 and 1.8 with HCl was inoculated with 1% (v / v) of Lactobacillus mucosa LM1 culture and then cultured at 37 ° C. After being sampled for 2 hours at the beginning of culture and at intervals of 30 minutes, the cells were diluted in sterilized physiological saline and plated on MRS plate medium. After culturing at 37 ° C for 48 hours to 72 hours, the number of colonies on the plate medium was counted, Were measured. As shown in FIG. 3, at pH 1.8, the number of viable cells gradually decreased with the lapse of incubation time. However, at pH 3 close to the above physiological pH, the number of bacteria did not substantially decrease even after 2 hours of culture as in the case of neutral pH control. This may be the basis for confirming that even if Lactobacillus mucosa LM1 lactic acid bacteria is ingested, it is excellent in acid resistance enough to withstand gastric pH.

2-2. My bile  Research

The biliary properties of Lactobacillus mucosa LM1 according to the present invention were examined. Considering that the intestinal bile acid concentration was around 0.1%, MRS liquid medium was prepared so that the concentrations of porcine bile salts were 0, 0.1, 0.3 and 0.5%, respectively. Lactobacillus mucosa LM1 according to the present invention was inoculated into a sterilized MRS liquid medium and cultured at 37 ° C for 16-24 hours, and then 0.1% (v / v) was inoculated into the MRS liquid medium containing bile salt . After inoculation, samples were collected every 30 minutes while culturing at 37 ° C, and then diluted in sterilized physiological saline, and plated on MRS plate medium. After incubation at 37 ° C for 48 hours to 72 hours, the number of colonies And the viable cell count was measured. As shown in FIG. 4, the bacterium number was rapidly reduced in the high concentration of bile salt (0.5%) only for 30 minutes after the incubation. However, since the lactic acid bacterium was maintained at a proper concentration of 0.3% It can be a basis for confirming that the LM1 can survive sufficiently in human or animal fields.

Example  3. Lactobacillus Mukose LM1 ( Lactobacillus mucosae LM1 ) Minister of Adsorption capacity  Research

The lactic acid bacteria must have a high survival rate in the intestinal tract in order to effectively effect a rectal effect in the intestines, and the intestinal mucosal adsorption ability should be excellent for a high survival rate. Lactobacillus can also be used to control the immune system of humans and animals through intestinal epithelial cells after adsorption to the intestinal tract.

Mucin adhesion, hydrophobicity, auto-aggregation, and co-aggregation with pathogenic bacteria have been widely used for the analysis of lactic acid bacteria in the intestinal tract. And in the present invention, the intestinal adsorptive capacity of Lactobacillus mucosa LM1 was compared with other strains using several methods described above.

3-1. Lactobacillus Mukose LM1 And other strains Mushin Adsorption capacity ( mucin adhesion ) compare

Lactobacillus lucosus GG (LGG bacteria), which is excellent in absorption capacity of intestinal tract and used as a yoghurt bacterium worldwide, was examined for the intestinal adsorption of Lactobacillus mucosa LM1.

Porcine mucin (type III, bound sialic acid 0.5-1.5%) from Sigma-Aldrich (USA) was used. The strains used in the experiment were incubated at 37 ° C for 12 hours and then used in the experiment. The intestinal mucosal capacity was 100 μl of mucin at a concentration of 1-10 μg / ml in a 96-well plate, and the mixture was stirred at 4 ° C. for 1 hour to uniformly coat the wells. The remaining untreated mucin solution was carefully removed with a pipet and stored at 4 ° C.

The strains to be used for the experiment were cultured in the MRS liquid medium for 24 hours to 48 hours, and then the cells were recovered by centrifugation at 3,000 × g. The recovered cells were washed twice with phosphate buffer (10 mM, pH 7.0) and suspended in phosphate buffer at a concentration of 5.0 × 10 ⁸ / μl to 9.0 × 10 ⁸ / μl. 200 μl of the suspension solution was added to a mucin-coated well and incubated at 37 ° C for 30 minutes to 2 hours. Then, the culture solution not adsorbed to mucin was removed by pipette. After 200 μl of Triton-X 100 (0.5% v / v) was injected into the wells, the wells were washed three times with sterilized citrate buffer to remove mucin The adsorption strain was recovered. After recovering the recovered strain, the strain was plated on MRS plate medium and cultured at 37 ° C. for 24 to 48 hours. The number of strains suspended in the phosphate buffer solution was regarded as 100%, and the number of lactic acid bacteria adsorbed on the mucins was expressed in terms of% in FIG. As a result, LGG strain showed a mucin adsorption rate of 54.5%, and Lactobacillus mucosa LM1 according to the present invention had 84.8%, showing the best mucin adsorption rate among comparative strains (FIG. 5).

3-2. Lactobacillus Mukose LM1 Of pathogenic bacteria Cohesion ability ( co - aggregation ) compare

Lactic acid bacteria is a very important factor for suppressing pathogenic bacteria in the intestines and cohesion with pathogenic bacteria. In the present invention directed to Salmonella bacteria (indicator strain) typhimurium (Salmonella typhimurium KCCM 40253 strain was used.

Lactobacillus mucosa LM1 and salmonella were inoculated into MRS liquid medium and Nutrient liquid medium (Difco, USA), respectively, and cultured at 37 ° C for 24 hours to 48 hours. Then, the cells were recovered by centrifugation at 10,000 × g for 15 minutes Respectively. This was suspended again in a PBS buffer to adjust the viable cell count to a level of 1.0 × 10 ⁸ / mL to 5.0 × 10 ⁸ / mL. Lactobacillus mucosa LM1 and salmonella suspension were each mixed at 2 ml each. The mixture was stirred for 10 seconds, then stored at 37 ° C for 24 hours, and then coagulated for 2, 16, 20 and 24 hours. Co-aggregation was calculated according to the method of Kos et al. (2003) as follows.

- A _X : OD ₆₀₀ of Lactobacillus muocet LM1 suspension

- A _Y : OD ₆₀₀ of the salmonella suspension

- A _{X + Y} : OD ₆₀₀ of Lactobacillus mucosa LM1 and salmonella mixture

As shown in FIG. 6, the agglutination rate of the lactic acid bacteria used in the experiment gradually increased with the incubation time. After 24 hours of incubation, the LGG lactic acid bacteria exhibited a coagulation rate of 24.0% with Salmonella, and Lactobacillus Johnson PF01 johnsonii PF01) showed 26.8% of salmonella coagulation. On the other hand, Lactobacillus mucosa LM1 according to the present invention exhibited an aggregation ratio of 36.5% with Salmonella, and exhibited a better agglutination rate than other strains known to have excellent intestinal absorption ability.

3-3. Lactobacillus Mukose LM1  Hydrophobicity of cell surface ( hydrophobicity ) Research

Because the cell surface hydrophobicity is highly correlated with the adsorption rate of the intestinal tract, the organoleptic capacity of the bacteria can be investigated (Munoz-Provecio et al., 2009). Generally, hexadecane is used as a solvent for measuring the degree of hydrophobicity of the cell surface. In the present invention, the cell wall hydrophobicity of Lactobacillus mucosa LM1 was also compared with other strains using the organic solvent. Lactobacillus mucosa LM1 was inoculated in 10 ml of MRS liquid medium, and cultured at 37 ° C for 24 to 48 hours, followed by centrifugation at 10,000 xg for 15 minutes to recover the cells. This was suspended in PBS buffer and adjusted to OD ₆₀₀ (turbidity at ₆₀₀ nm wavelength) to about 0.4 (= A ₀ ). 3 ml of the suspension was mixed with 1 ml of hexa tencene and stirred for 1 minute and left for 20 minutes so that the aquous phase and the solvent phase were separated from each other. 1 ml of the upper layer of the water layer was carefully taken and OD ₆₀₀ was measured and calculated as follows.

- A0: Initial OD ₆₀₀

- A1: After reacting with hexaethene, OD _{600 of} water layer

As shown in FIG. 7, the cell surface hydrophobicity of LGG strain was 42%, while the cell surface hydrophobicity of Lactobacillus mucosa LM1 according to the present invention was 74%, which is very higher than other strains. It was confirmed that the intestinal absorption rate of Lactobacillus mucosa LM1 strain was very high.

3-4. Lactobacillus Mukose LM1 Of harmful bacteria Inhibition  Research

Probiotics Lactic acid bacteria are known to inhibit the growth of other microorganisms by lowering the surrounding pH by producing organic acids such as lactic acid, and to produce antimicrobial substances such as bacteriocin depending on the strain.

Therefore, the antibacterial activity against the Escherichia coli Lactobacillus mucosa LM1 of the present invention against E. coli K88 was first examined. Lactobacillus mucosa LM1 was inoculated into MRS broth medium (Difico, USA) and cultured at 37 ° C for 24 hours and 36 hours. The culture broth was centrifuged at 10,000 rpm for 10 minutes to remove the cells. In order to eliminate the antimicrobial activity due to the low pH of the culture solution from which the cells were removed, ammonium sulfate was added to the culture solution at a concentration of 80% to precipitate the proteinaceous component. The precipitated protein components were recovered by centrifugation at 15,000 rpm for 10 minutes, followed by dialysis with 50 mM phosphate buffer (pH 7.0) to remove salts. Protein components were recovered and diluted before use in the antimicrobial activity test. After punching holes in the Tryptic soy solid medium with the indicator bacteria (Escherichia coli) plated, 200 μl of the protein solution recovered from the 24 hour and 36 hour culture medium of Lactobacillus mucosa LM1 was added and cultured at 37 ° C for 18 hours. As a result, E. coli growth inhibition rings were observed in both the 24-hour and 36-hour culture concentrates (FIG. 8), indicating that Lactobacillus LM1 lactic acid bacteria produced proteinaceous antimicrobial substances.

Next, we examined whether or not other harmful bacteria were inhibited in addition to E. coli. The indicator bacteria used in the experiment were E. coli , Salmonella ( Salmonella spp. enteritidis ) and gram-negative bacteria including Staphylococcus Gram-positive bacteria such as aureus , and microorganisms ranging from fungi were used as indicator bacteria.

As a result of investigating the inhibition of indicator bacteria by the same method (agar diffusion method) as that of E. coli, Salmonella typhimurium , Pseudomonas aeruginosa ), and Listeria monocytogenes ( Listeria monocytogenes , Staphylococcus aureus , and Streptococus mutans , or fungi such as Candida albicans .

Therefore, the proteinaceous antimicrobial substance secreted by Lactobacillus Lactobacillus LM1 lactic acid bacteria itself or Lactobacillus Lactobacillus LM1 lactic acid bacteria may be useful for inhibiting Gram negative bacteria such as Escherichia coli, which is involved in human and animal diseases.

Harmful bacteria inhibition spectrum of Lactobacillus mucosa LM1 Indicator Inhibition Escherichia coli K88 + Salmonella typhimurium KCCM 40253 + Salmonella enteritidis KCCM 12021 - Klebsiella pneumoniae KCCM 11418 - Pseudomonas aeruginosa KCCM 11266 + Listeria monocytogenes ATCC 19114 - Streptococcus agalactiae ATCC 13813 - Streptococcus mutans ATCC 25175 - Staphylococcus aureus KCCM 40510 - Enterococcus faecalis VRE CCARM 0011 - Bacillus cereus KCTC 1661 + Candida albicans KCTC 7122 -

Example  4. Lactobacillus Mukose LM1 Use of

The present invention also provides foods, medicaments and feed additives containing Lactobacillus mucosa LM1 lactic acid bacteria, which are superior to conventional lactic acid bacteria used as probiotics, in terms of absorption capacity, fixation rate and survival rate.

When the lactobacillus mucosa LM1 lactic acid bacterium of the present invention is used as a food additive, the above-mentioned lactic acid bacterium may be directly added or used together with other food or food ingredients, and may be suitably used according to a conventional method. Examples of the food to which the lactic acid bacteria can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, and it is preferably prepared from fermented milk, yogurt, cheese, milk beverage or milk powder, more preferably a food additive or a health supplement. The amount of the active ingredient to be mixed can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). Generally, the lactobacillus mucosa LM1 lactic acid bacterium of the present invention is added to 1 x 10 ⁷ to 10 x 10 ⁸ (bacterium) of the present invention per 1 g (or ml) Preferably in an amount of 1 × 10 ⁸ to 5 × 10 ⁸ . The effective dose of the strain of the present invention may be used in accordance with the effective dose of the composition, but may be less than the above range for health and hygiene purposes or long-term intake intended for health control, It can be used in an amount exceeding the range described above.

When a composition comprising the lactobacillus mucosa LM1 lactic acid bacterium of the present invention is used for the production of a pharmaceutical composition, it may further comprise a suitable carrier, excipient or diluent commonly used. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The composition containing Lactobacillus mucosa LM1 according to the present invention may be formulated into oral compositions such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterilized injection solutions And the like. More specifically, it can be prepared by using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, etc., which is generally used in the case of formulation.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ) Or lactose, gelatin and the like. Lubricants such as magnesium stearate and talc other than simple excipients may also be used.

Examples of liquid formulations for oral use include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

When the lactobacillus mucosa LM1 lactic acid bacterium of the present invention is used as a feed additive, the lactic acid bacterium may be used as it is, or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The feed additive of the present invention may be added with additives such as known carriers or stabilizers which are acceptable for pharmaceutical, food or feed as well as the above-mentioned lactic acid bacteria. If necessary, various feeds such as vitamins, amino acids and minerals, antioxidants, antibiotics , An antibacterial agent and other additives may be added, and the form thereof may be a proper state such as powder, granule, pellet, suspension and the like. When the feed additive of the present invention is supplied, it can be supplied singly or mixed with feed.

When the lactobacillus mucosa LM1 lactic acid bacterium of the present invention is used as a preparation or a prodrug, the composition may contain the lactic acid bacterium itself or a culture of the lactic acid bacterium as an active ingredient, and may further contain an excipient or a carrier. have. The composition includes a culture medium itself cultured in a liquid medium, a filtrate obtained by removing the strain by filtration or centrifugation of the culture medium (centrifuged supernatant), and the like. The content of the Lactobacillus mucosa LM1 strain in the composition may vary depending on the use of the composition and the formulation. The dressing or probiotic composition according to the present invention can be manufactured and administered in a variety of formulations and methods. For example, the culture of Lactobacillus < RTI ID = 0.0 > Mucosa LM1, < / RTI > is mixed with a carrier and flavoring commonly used in the pharmaceutical field to form tablets, troche, capsules, elixir, May be formulated and administered in the form of a syrup, powder, suspension, granule, or the like. Examples of the carrier include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents and the like. The administration method may be oral, parenteral or application, but is preferably orally administered. The administration dose can be appropriately selected according to the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, age, sex, condition, etc. of the subject.

Korea Microorganism Conservation Center (overseas) KCCM11442P 20130809

<110> DANKOOK INDUSTRY-ACADEMIC COOPERATION FOUNDATION <120> Lactobacillus mucosae LM1 with excellent acid-Resistant,          bile-resistance, antibacterialactivity and adhere to mucosal          surfaces, and composition containing the same <130> P13-0037 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1456 <212> DNA <213> Lactobacillus mucosae LM1 <400> 1 agtcgacgcg ttggcccaac tgattgaacg tgcttgcacg gacttgacgt tggtttacca 60 gcgagtggcg gacgggtgag taacacgtag gtaacctgcc ccaaagcggg ggataacatt 120 tggaaacaga tgctaatacc gcataacaat ttgaatcgca tgattcaaat ttaaaagatg 180 gcttcggcta tcactttggg atggacctgc ggcgcattag cttgttggta gggtaacggc 240 ctaccaaggc tgtgatgcgt agccgagttg agagactgat cggccacaat ggaactgaga 300 cacggtccat actcctacgg gaggcagcag tagggaatct tccacaatgg gcgcaagcct 360 gatggagcaa caccgcgtga gtgaagaagg gtttcggctc gtaaagctct gttgttagag 420 aagaacgtgc gtgagagcaa ctgttcacgc agtgacggta tctaaccaga aagtcacggc 480 taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttatccg gatttattgg 540 gcgtaaagcg agcgcaggcg gtttgataag tctgatgtga aagcctttgg cttaaccaaa 600 gaagtgcatc ggaaactgtc agacttgagt gcagaagagg acagtggaac tccatgtgta 660 gcggtggaat gcgtagatat atggaagaac accagtggcg aaggcggctg tctggtctgc 720 aactgacgct gaggctcgaa agcatgggta gcgaacagga ttagataccc tggtagtcca 780 tgccgtaaac gatgagtgct aggtgttgga gggtttccgc ccttcagtgc cgcagctaac 840 gcattaagca ctccgcctgg ggagtacgac cgcaaggttg aaactcaaag gaattgacgg 900 gggcccgcac aagcggtgga gcatgtggtt taattcgaag ctacgcgaag aaccttacca 960 ggtcttgaca tcttgcgcca accctagaga tagggcgttt ccttcgggaa cgcaatgaca 1020 ggtggtgcat ggtcgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080 cgcaaccctt gttactagtt gccagcattc agttgggcac tctagtgaga ctgccggtga 1140 caaaccggag gaaggtgggg acgacgtcag atcatcatgc cccttatgac ctgggctaca 1200 cacgtgctac aatggacggt acaacgagtc gcgaactcgc gagggcaagc taatctctta 1260 aaaccgttct cagttcggac tgcaggctgc aactcgcctg cacgaagtcg gaatcgctag 1320 taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380 acaccatgag agtttgcaac acccaaagtc ggtggggtaa cccttcgggg agctagccgc 1440 ctaaggtggg gcagat 1456

Claims (9)

Acid, bile star, excellent Secretary of adsorption capacity and E. coli (Escherichia coli), rat typhoid bacteria (Salmonella typhimurium), Pseudomonas aeruginosa (Pseudomonas aeruginosa) and Selena funny bacteria (Bacillus cereus) suppression of the superior Lactobacillus Mu kose LM1 on ( Lactobacillus mucosae LM1) KCCM11442P. The method according to claim 1,
The Lactobacillus mucosa LM1 ( Lactobacillus mucosae LM1) LM1 KCCM11442P is Lactobacillus mu kose (Lactobacillus mucosae LM1) KCCM11442P, characterized in that consisting of the nucleotide sequence of SEQ ID NO: 1. Acid, bile star, excellent Secretary of adsorption capacity and E. coli (Escherichia coli), rat typhoid bacteria (Salmonella typhimurium), Pseudomonas aeruginosa (Pseudomonas aeruginosa) and Selena funny bacteria (Bacillus cereus) suppression of the superior Lactobacillus Mu kose LM1 on ( Lactobacillus mucosae LM1) Culture medium of KCCM11442P. ( Lactobacillus mucosae LM1) KCCM11442P or a culture thereof having excellent inhibitory ability against Escherichia coli , Salmonella typhimurium , Pseudomonas aeruginosa and Bacillus cereus . Wherein the antimicrobial composition delete Lactobacillus LM1 ( Lactobacillus mucosae LM1) KCCM11442P or a culture thereof. Lactobacillus LM1 ( Lactobacillus mucosae LM1) KCCM11442P or a culture thereof. Lactobacillus LM1 ( Lactobacillus mucosae LM1) KCCM11442P or a culture thereof. Lactobacillus LM1 ( Lactobacillus mucosae LM1) KCCM11442P or a culture thereof.

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