KR101772870B1 - Novel Lactobacillus plantarum with probiotic activities and use thereof - Google Patents

Novel Lactobacillus plantarum with probiotic activities and use thereof Download PDF

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KR101772870B1
KR101772870B1 KR1020160015171A KR20160015171A KR101772870B1 KR 101772870 B1 KR101772870 B1 KR 101772870B1 KR 1020160015171 A KR1020160015171 A KR 1020160015171A KR 20160015171 A KR20160015171 A KR 20160015171A KR 101772870 B1 KR101772870 B1 KR 101772870B1
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lactobacillus plantarum
mjm60319
mjm60399
mjm60298
strain
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KR1020160015171A
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KR20170093586A (en
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서주원
양승환
사시쿠마
다모다란 카티
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명지대학교 산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/32—Foods, ingredients or supplements having a functional effect on health having an effect on the health of the digestive tract
    • A23V2200/3204—Probiotics, living bacteria to be ingested for action in the digestive tract
    • A23Y2220/67—
    • C12R1/25—

Abstract

The present invention provides a novel Lactobacillus plantarum strain having probiotic activity. The Lactobacillus plantarum strain according to the present invention is excellent in acid resistance, bile acidity or adherent epithelial cell adhesion, and can reach the most part when it is orally administered to a subject, and the propagation of harmful bacteria in the intestines It is useful as a probiotic material because it can be effectively inhibited. In addition, the Lactobacillus plantarum strain according to the present invention has excellent antimicrobial activity and is excellent in β-galactosidase activity to prevent or ameliorate lactose intolerance It can be used as a useful pharmaceutical material. In addition, the specific Lactobacillus plantarum strain according to the present invention is excellent in survival stability in a sodium chloride-resistant or lyophilized process and can maintain a high diverse physiological activity even when it is formulated into a pharmaceutical drug.

Description

Novel Lactobacillus plantarum with probiotic activity and use thereof < RTI ID = 0.0 >

More particularly, the present invention relates to a novel lactic acid bacterium which is isolated from a fermented cow milk curd, which is a traditional Indian food, and is safe for human body, and has an activity required for probiotics such as acid resistance, bile acidity, And relates to novel Lactobacillus plantarum having various functions such as antimicrobial activity, beta -galactosidase activity, sodium chloride resistance, storage stability and the like and pharmaceutical uses thereof.

Humans are living in constant contact with microorganisms. Numerous microorganisms exist inside and outside the human body, which are beneficial to human health, and sometimes cause diseases. These microorganisms are called overgrown bacteria. Most of the gut bacteria in the body are present in the gastrointestinal tract. The mass of microorganisms that coexist with humans is over 1kg in total, and the number is about 100 trillion or more. These microbial populations are called intestinal flora and they form an ecosystem within the intestinal tract. The roles of intestinal flora as well as metabolism, nutrition, energy utilization, regulation of innate immunity and adaptive immunity, development and production of intestinal mucosal epithelium and prevention of intrusion of pathogens are known. The intestinal flora has significant differences in microbial species and strains depending on the individual. The microorganisms originated from the mother, the genotype of the host, and the unique environment of the intestinal flora are shown. In addition, intestinal flora is affected by dietary habits, age, medications, surgery, radiation therapy, stress, and diseases the host has. Because of the variable factors affecting intestinal bacterial flora, it is not easy to determine the relationship with disease, but irritable bowel syndrome, inflammatory bowel disease, obesity, constipation, fatty liver, neonatal necrotizing enteritis and allergic diseases are related to intestinal flora .

Probiotics is a living microorganism that is beneficial to the health of the host when administered in sufficient amounts. Most probiotics known so far are mostly of the kind we know to be lactic acid bacteria. In order to be recognized as probiotics, it should not reach the small intestine, but should proliferate and settle in the intestine. It should have a beneficial effect in the intestine, and should not be toxic or pathogenic. Recently, there is a tendency to try to use probiotics in the area of colonic diseases such as irritable bowel syndrome, antibiotic-associated diarrhea, inflammatory bowel disease, and constipation. Irritable bowel syndrome or antibiotic-associated diarrhea is associated with the use of antibiotics or a history of infectious enteritis that causes changes in the composition of intestinal flora. Therefore, in these cases, attempts have been made to restore the intestinal flora to normal using probiotics. As a representative probiotic, there is Lactobacillus sp. Lactobacillus . Microorganisms of the genus Lactobacillus are lactic acid bacterium which are homozygous or heterozygous fermenting, and are commonly found in the fermentation process of minced meat, dairy products and vegetables of animals including humans. Lactobacillus microorganisms maintain intestinal pH acidic and inhibit the growth of harmful bacteria such as E. coli and Clostridium, and improve diarrhea and constipation, as well as vitamin synthesis, anticancer activity, serum cholesterol lowering [Michael and Philippe, Probiotics and prebiotics: Effects on diarrhea, The journal of nutrition, Volume 137, March 2007, pages 803S-811S; Roberfroid, Prebiotics and probiotics: Are they functional foods ?, American Journal of Clinical Nutrition, Volume 71, June 2000, pages 1682S-1687S].

As the recent income level increases, the intake of high-calorie foods increases in the dietary culture, and the incidence of obesity and various adult diseases is increasing. Among the factors causing adult diseases, the most important factor is high blood cholesterol level due to excessive intake of animal fat, which is a risk index for diseases such as hyperlipidemia, atherosclerosis and lipid-related metabolic syndrome. Based on many studies on the selection of lactic acid bacteria that degrade cholesterol, the mechanism by which the lactic acid bacteria decrease the cholesterol absorption can be generally explained in three ways (see Korean Patent No. 10-0930251). First, lactic acid bacteria colonize the intestinal tract and cholesterol is adsorbed and excreted. Second, lactic acid bacteria adsorb and excrete bile acids. In this case, cholesterol is a precursor of bile acid, so it can use cholesterol to supplement deficient bile acid, which can consequently reduce intestinal cholesterol concentration. And the third mechanism is the action of lactic acid bacteria to decouple conjugated bile acid. The free bile acid produced in the liver binds to taurine or glycine in the small intestine and becomes a conjugated bile acid. Conjugated bile acid has high lipid solubility and facilitates the absorption of lipids such as cholesterol in the small intestine. Therefore, in the case of large amounts of cholesterol, the amount of cholesterol absorbed into the blood increases and the blood cholesterol concentration increases. However, deconjugated bile acid has a low lipid solubility and acts to lower lipid lipid transporters such as cholesterol. Therefore, a strain having a bile salt hydrolase that de-capsulates a conjugated bile acid may lower lipid solubility and lower the absorption of cholesterol in the small intestine. Therefore, it is necessary to develop a strains which are safe for human body and excellent in cholesterol-lowering function, and to be applied as medicines or functional foods. Korean Patent Registration No. 10-0774969 discloses Lactobacillus plantarum that can inhibit cholesterol absorption in the intestinal tract, increase HDL lipoprotein level in the blood, lower LDL lipoprotein, and prevent arteriosclerosis by improving blood lipids. In addition, U.S. Published Patent Application No. 20110117629 discloses Lactobacillus plantarum BB9 capable of attaching to the gastrointestinal tract and removing cholesterol. However, most Lactobacillus sp. Lactic acid bacteria are known to have a low cholesterol level when they are orally administered to the human body due to insufficient acid resistance or biliary cholesterol.

Gastrointestinal infection caused by intestinal pathogens is one of the most important problems in animal husbandry. The most prevalent intestinal pathogenic infections are enterotoxigenic Escherichea coli infection, enteropathogenic Escherichia coli coli), Salmonella, and the like Entebbe Rica (Salmonella enterica) infection in poultry Salmonella Galina Room (Salmonella gallinarum) infection, from gastroenteritis (gastroenteritis) caused the condition leading to systemic infection and induce diarrhea in young pigs and calves. Antibiotics are widely used to prevent infection or promote growth of intestinal pathogens in young livestock. However, since antibiotics are resistant to antibiotics due to the abuse and / or abuse of antibiotics, alternative strategies are needed to control intestinal pathogen infection. Probiotics have emerged as an alternative to antibiotics, and probiotics can inhibit bacterial colonization of intestinal pathogens, stimulate host defense, and enhance intestinal barrier function. Preventing the adherence of pathogenic bacteria to intestinal cells is an important step in preventing pathogen mycosis and pathogen infection, which can be achieved by the use of probiotics.

It is important to select probiotic lactic acid bacteria based on gastrointestinal tolerance. In order for the probiotic bacteria to be effective, they must be able to survive in the intestinal environment and adhere to intestinal epithelial cells and inhabit large quantities in the intestines [Salminen, S. et al., (1996) Clinical uses of probiotics for stabilizing the gut mucosal barrier: successful strains and future challenges. Antonie Van Leeuwenhoek 70: 347-358; Conway, P. (1996) Selection criteria forprobiotic microorganisms. Asia Pacific J Clin Nutr 5: 10-14]. In addition, safety and pathogenicity testing is required for lactic acid bacteria selected from new sources to be used for probiotic use [Holzapfel and Schillinger 2002; Ji et al. 2013]. Specifically, microorganisms administered to humans or livestock should not produce biogenic amines to use and. Biogenic amines are known not to be inactivated by heat treatment. Also, in order to be used for probiotics use, lactic acid bacteria should not destroy mucosal barriers and should not break micin. The disruption of the mucosal barrier and degradation of the intestinal mucin may cause bacteremia if internal organs are displaced (Zhou et al., 2001). Also, in order to be used for probiotics use, lactic acid bacteria should not possess inherited drug resistance genes. Administration of lactic acid bacteria with inherited drug resistance genes can lead to the development of intestinal microorganisms with drug resistance (Sharma et al. 2014). However, most lactic acid bacteria are known to have inherent resistance to various antibiotics [Saarela et al. 2000; Argyri et al. 2013]. In addition to safety, microorganisms used as probiotics should be able to survive in processing. Lactobacillus cultures for use as probiotics are generally lyophilized and processed into the form of a powder present in the capsule. A cryoprotectant may be used to stabilize the membrane integrity and minimize degradation of lactic acid bacteria during freeze-drying of lactic acid bacteria cultures [Zarate and Nader-Macias 2006; Forssten et al. 2011].

It is an object of the present invention to provide a new lactic acid bacteria which is safe as a foodstuff having various functions beneficial to the human body together with activity required as probiotics.

It is also an object of the present invention to provide various uses of novel lactic acid bacteria.

The inventors of the present invention have found that a myriad of lactic acid bacteria are screened from fermented cow milk curd, which is a traditional food of India, and a specific Lactobacillus plantarum strain is safe as a foodstuff and at the same time, Having various useful functions, and completed the present invention.

In order to solve one object of the present invention, an example of the present invention is a 16S rDNA comprising the nucleotide sequence as set forth in SEQ ID NO: 1 and having an acid resistance, bile acid resistance, adherent epithelial cell, antimicrobial activity, beta-galactosidase The present invention provides a Lactobacillus plantarum strain characterized by having at least one property selected from the group consisting of β-galactosidase activity, bile salt hydrolase activity and sodium chloride resistance.

In order to solve one object of the present invention, an example of the present invention provides a probiotic composition comprising Lactobacillus plantarum MJM60319 (accession number: KACC 92111P) or a culture thereof as an active ingredient .

In order to solve one object of the present invention, an example of the present invention is an Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum MJM60298 (accession number: KACC 92110P ) And Lactobacillus plantarum MJM60399 (accession number: KACC 92112P) or a cultured product of said mixed lactic acid bacteria as an active ingredient .

In order to solve one object of the present invention, an example of the present invention relates to an antibacterial agent comprising Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), a culture thereof, a crushed product thereof or an extract thereof as an active ingredient ≪ / RTI >

In order to solve one object of the present invention, an example of the present invention is an Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum MJM60298 (accession number: KACC 92110P ), And Lactobacillus plantarum MJM60399 (Accession No .: KACC 92112P), a culture of the mixed lactic acid bacteria, a disruption of the mixed lactic acid bacteria or an extract of the mixed lactic acid bacteria, As an active ingredient.

In order to solve one object of the present invention, an example of the present invention relates to a method for producing Lactobacillus plantarum including Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), a culture thereof, a lysate thereof or an extract thereof, A composition for preventing or ameliorating lactose intolerance is provided.

In order to solve one object of the present invention, an example of the present invention is an Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum MJM60298 (accession number: KACC 92110P ), And Lactobacillus plantarum MJM60399 (Accession No .: KACC 92112P), a culture of the mixed lactic acid bacteria, a disruption of the mixed lactic acid bacteria or an extract of the mixed lactic acid bacteria, A composition for preventing or ameliorating lactose intolerance contained as an active ingredient.

The Lactobacillus plantarum strain according to the present invention is excellent in acid resistance, bile acidity or adherent epithelial cell adhesion, and can reach the most part when it is orally administered to a subject, and the propagation of harmful bacteria in the intestines It is useful as a probiotic material because it can be effectively inhibited. In addition, the Lactobacillus plantarum strain according to the present invention has excellent antimicrobial activity and is excellent in β-galactosidase activity to prevent or ameliorate lactose intolerance It can be used as a useful pharmaceutical material. In addition, the specific Lactobacillus plantarum strain according to the present invention is excellent in survival stability in a sodium chloride-resistant or lyophilized process and can maintain a high diverse physiological activity even when it is formulated into a pharmaceutical drug.

Figure 1 shows the morphology of the finally selected strains MJM60319, MJM60298 and MJM60399.
Figure 2 is a diagram showing the phylogenetic tree of strains MJM60319, MJM60298 and MJM60399.
FIG. 3 is a graph showing the results of measurement of the production of biologically-derived amines of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
Fig. 4 is a diagram showing the results of measurement of mucin decomposition of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
FIG. 5 is a graph showing the results of measurement of hemolytic activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
6 is a diagram showing the results of measurement of antimicrobial activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
FIG. 7 is a graph showing the results of resistance evaluation of Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 under oral-gastrointestinal transit conditions.
8 is a photograph showing the adherence ability of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 to Caco-2 cell monolayer.
9 is a photograph showing the results of measurement of beta -galactosidase activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
10 is a graph showing the growth levels of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 under various sodium chloride concentrations.
11 is a graph showing the production ability of D, L-lactate of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.
12 is a graph showing the results of measurement of the survival stability of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 under lyophilization conditions.

Hereinafter, terms used in the present invention will be described.

The term "culture product " used in the present invention means a product obtained by culturing a microorganism in a known liquid medium or solid medium, and includes a microorganism.

The terms " pharmaceutically acceptable "and" pharmaceutically acceptable "in the present invention are intended to mean not significantly irritating the organism and not interfering with the biological activity and properties of the administered active substance.

As used herein, the term "prophylactic " means any act that inhibits the symptoms of a particular disease or delays its progress by administration of the composition of the present invention.

The term "treatment" as used herein refers to any action that improves or alleviates the symptoms of a particular disease upon administration of the composition of the present invention.

The term "improvement" as used in the present invention means all actions that at least reduce the degree of symptom associated with the condition being treated.

The term "administering" as used herein is meant to provide any desired composition of the invention to an individual by any suitable method. The term individual refers to any animal such as a human, a monkey, a dog, a goat, a pig, or a mouse having a disease in which symptoms of a specific disease can be improved by administering the composition of the present invention.

The term " pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit or risk rate applicable to medical treatment, including the type of disease, severity, activity of the drug, The time of administration, the route and rate of excretion of the drug, the duration of the treatment, factors including drugs used simultaneously and other factors well known in the medical arts.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to a novel lactobacillus that is pharmaceutically safe and has various functions beneficial to the human body together with the activity required as probiotics.

A novel lactic acid bacterium according to the present invention is a 16S rDNA comprising Lactobacillus plantarum containing the nucleotide sequence of SEQ ID NO: 1, Lactobacillus plantarum containing the nucleotide sequence of SEQ ID NO: 2 as 16S rDNA, ( Lactobacillus plantarum ) or a 16S rDNA selected from Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 3. Preferably, the lactic acid bacteria according to the present invention are selected from the group consisting of acid tolerance, bile acid resistance, intestinal epithelial cell adhesion, antibacterial activity, beta -galactosidase activity, bile acid salt hydrolase activity, And stability of survival in the process of freezing or lyophilizing, and the like. In addition, Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 1 as the 16S rDNA is preferably Lactobacillus plantarum MJM60319 (accession number: KACC 92111P). The Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 2 as the 16S rDNA is preferably Lactobacillus plantarum MJM60298 (accession number: KACC 92110P). The Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 3 as the 16S rDNA is preferably Lactobacillus plantarum MJM60399 (accession number: KACC 92112P). remind

Another aspect of the present invention relates to various uses of novel lactic acid bacteria. The present invention provides a probiotic composition, a composition for antimicrobial use, or a composition for preventing or improving lactose intolerance, for various uses of novel lactic acid bacteria. In the present invention, a probiotic composition refers to a microorganism-utilizing product capable of promoting health and promoting health of a living body, and usually includes a product group including a prophylactic agent or a probiotic agent.

The probiotic composition according to one embodiment of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum or a culture thereof containing the nucleotide sequence shown in SEQ ID NO: 1. In addition, the probiotic composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60319 (accession number: KACC 92111P) or a culture thereof as an active ingredient.

The probiotic composition according to an embodiment of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum or a culture thereof containing the nucleotide sequence shown in SEQ ID NO: 2. In addition, the probiotic composition according to one example of the present invention preferably contains Lactobacillus plantarum MJM60298 (accession number: KACC 92110P) or a culture thereof as an active ingredient.

The probiotic composition according to one embodiment of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 3 or a culture thereof. The probiotic composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60399 (accession number: KACC 92112P) or a culture thereof as an active ingredient.

The probiotic composition according to an example of the present invention is a 16S rDNA comprising Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 1, Lactobacillus plantarum containing 16S rDNA, Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: Planta column (Lactobacillus plantarum) and 16S rDNA as Lactobacillus Planta column (Lactobacillus plantarum) a culture of a second mixture of lactic acid bacteria or a mixture of lactic acid bacteria or more kinds selected from the group consisting of available, including the nucleotide sequence set forth in SEQ ID NO: 3 As an ingredient. The probiotic composition according to one embodiment of the present invention is preferably selected from the group consisting of Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum MJM60298 (accession number: KACC 92110P) and Lactobacillus plantarum MJM60399 (accession number: KACC 92112P), or a culture of the mixed lactic acid bacteria as an active ingredient.

Probiotic composition of the invention East agent which can improve the survival stability in the process of formulating the specific Lactobacillus Planta column (Lactobacillus plantarum) strain was lyophilized to culture medium of certain Lactobacillus Planta column (Lactobacillus plantarum) strains or a cryoprotectant. Examples of the cryoprotectant include whey protein concentrate, soy protein concentrate, skim milk powder or maltodextrin powder. The probiotic composition of the present invention preferably further comprises a skim milk powder and a maltodextrin powder.

The culture of lactic acid bacteria in the present invention is a product obtained by culturing a strain of Lactobacillus plantarum in a medium, which may be selected from known liquid or solid medium, for example, MRS liquid medium , MRS agar medium.

The probiotic composition of the present invention can be used as an active ingredient in the selection of acid resistance, bile acid resistance, adherent epithelial cell adhesion, antibacterial activity, beta -galactosidase activity, bile salt hydrolase activity or sodium chloride resistance ( Lactobacillus plantarum ) as an active ingredient, it is possible to improve normalization of intestinal bacterial flora, maintenance of intestinal health, improvement of immune function, dressing action by inhibition of intestinal harmful bacteria, lactic acidosis lactose intolerance, prevention of constipation, reduction of blood cholesterol, irritable bowel syndrome, inflammatory bowel disease, obesity, constipation, fatty liver, allergic diseases and the like.

An antimicrobial composition according to an example of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 1, a culture thereof, a crushed product thereof or an extract thereof. In addition, the antimicrobial composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), a culture thereof, a crushed product thereof or an extract thereof.

An antimicrobial composition according to an embodiment of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum containing the nucleotide sequence of SEQ ID NO: 2, a culture thereof, a crushed product thereof or an extract thereof. The antimicrobial composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60298 (accession number: KACC 92110P), a culture thereof, a crushed product thereof or an extract thereof.

An antimicrobial composition according to an embodiment of the present invention includes 16S rDNA as an active ingredient of Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 3, a culture thereof, a crushed product thereof or an extract thereof. The antimicrobial composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60399 (accession number: KACC 92112P), a culture thereof, a crushed product thereof or an extract thereof.

The antimicrobial composition according to one embodiment of the present invention comprises 16S rDNA, Lactobacillus plantarum containing the nucleotide sequence of SEQ ID NO: 1, Lactobacillus plantarum containing 16S rDNA and Lactobacillus plantarum containing the nucleotide sequence of SEQ ID NO: lanthanide column (Lactobacillus plantarum), and the culture, the mixture of Lactobacillus Planta column (Lactobacillus plantarum) group of two or more mixed lactic acid bacteria, selected from the consisting of the mixture of lactic acid bacteria comprising a nucleotide sequence shown in SEQ ID NO: 3 by 16S rDNA A lysate of a lactic acid bacterium or an extract of the mixed lactic acid bacterium as an active ingredient. The antimicrobial composition according to one embodiment of the present invention is preferably selected from the group consisting of Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum MJM60298 (accession number: KACC 92110P ), And Lactobacillus plantarum MJM60399 (Accession No .: KACC 92112P), a culture of the mixed lactic acid bacteria, a disruption of the mixed lactic acid bacteria or an extract of the mixed lactic acid bacteria, It is included as an active ingredient.

The antimicrobial composition of the present invention can be used as an antidisruptant agent capable of improving survival stability in the process of preparing a specific Lactobacillus plantarum strain by lyophilizing a culture solution of a specific Lactobacillus plantarum strain cryoprotectant). Examples of the cryoprotectant include whey protein concentrate, soy protein concentrate, skim milk powder or maltodextrin powder. The antimicrobial composition of the present invention preferably further comprises a skim milk powder and a maltodextrin powder.

Since the antimicrobial composition of the present invention contains Lactobacillus plantarum having antimicrobial activity against various harmful bacteria, it can be used for the infection of harmful bacteria, the inflammation reaction caused by infection of harmful bacteria, Diseases caused by infection, decay caused by harmful bacteria, and the like. Examples of such harmful bacteria include Salmonella typhimurium , Salmonella choleraesuis , Salmonella gallinarum , Escherichia E. coli , Staphylococcus aureus, and the like.

The composition for preventing or ameliorating lactose intolerance according to one embodiment of the present invention may be a 16S rDNA, a Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 1, a culture thereof, Extract as an active ingredient. The composition for preventing or ameliorating lactose intolerance according to one embodiment of the present invention is preferably a lactobacillus plantarum MJM60319 (accession number: KACC 92111P), a culture thereof, Extract as an active ingredient.

The composition for preventing or ameliorating lactose intolerance according to an example of the present invention may be a 16S rDNA, a Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 2, a culture thereof, Extract as an active ingredient. The antimicrobial composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60298 (accession number: KACC 92110P), a culture thereof, a crushed product thereof or an extract thereof.

The composition for preventing or ameliorating lactose intolerance according to one embodiment of the present invention is a 16S rDNA comprising a Lactobacillus plantarum containing the nucleotide sequence shown in SEQ ID NO: 3, a culture thereof, Extract as an active ingredient. The antimicrobial composition according to one embodiment of the present invention preferably contains Lactobacillus plantarum MJM60399 (accession number: KACC 92112P), a culture thereof, a crushed product thereof or an extract thereof.

A composition for preventing or ameliorating lactose intolerance according to one embodiment of the present invention is a 16S rDNA comprising Lactobacillus plantarum , 16S rDNA comprising the nucleotide sequence shown in SEQ ID NO: 1, Lactobacillus Planta column (Lactobacillus plantarum) and 16S rDNA of SEQ ID NO: 2 or more species selected from Lactobacillus sample group consisting of a lanthanide column (Lactobacillus plantarum) comprising a base sequence described in the 3 mixed lactic acid bacteria, wherein comprising the nucleotide sequence A culture of a mixed lactic acid bacterium, a lysate of the mixed lactic acid bacteria or an extract of the mixed lactic acid bacteria as an active ingredient. The composition for preventing or ameliorating lactose intolerance according to one embodiment of the present invention is preferably Lactobacillus plantarum MJM60319 (accession number: KACC 92111P), Lactobacillus plantarum ), A mixture of two or more mixed lactic acid bacteria selected from the group consisting of MJM60298 (accession number: KACC 92110P) and Lactobacillus plantarum MJM60399 (accession number: KACC 92112P), a culture of the mixed lactic acid bacteria, Or an extract of the mixed lactic acid bacteria as an active ingredient.

The composition for preventing or ameliorating lactose intolerance according to the present invention is characterized in that in the process of preparing a specific Lactobacillus plantarum strain by lyophilizing a culture solution of a specific Lactobacillus plantarum strain, And a cryoprotectant capable of improving the cryoprotectant. Examples of the cryoprotectant include whey protein concentrate, soy protein concentrate, skim milk powder or maltodextrin powder. The composition for preventing or improving lactose intolerance of the present invention preferably further comprises a skim milk powder and a maltodextrin powder.

The composition for preventing or ameliorating lactose intolerance of the present invention contains Lactobacillus plantarum which is excellent in beta -galactosidase activity, so that lactose is hydrolyzed to form galactose And glucose.

In the present invention, the probiotic composition, the antibacterial composition, and the composition for preventing or improving lactose intolerance may be used as a pharmaceutical composition, a food additive, a food composition (especially a health functional food) or a feed additive Can be embodied. In addition, the content of the active ingredient, such as lactic acid bacteria, may be adjusted in various ranges depending on the specific form of the composition, the purpose of use, and the manner of use.

In the pharmaceutical composition according to the present invention, the content of the novel lactic acid bacteria, the culture thereof, the lysate thereof or the extract thereof is not particularly limited and may be 0.01 to 99% by weight, 50% by weight, more preferably 1 to 30% by weight. In addition, the pharmaceutical composition according to the present invention may further contain, in addition to the active ingredient, an additive such as a pharmaceutically acceptable carrier, excipient or diluent. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention 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. In addition, the pharmaceutical compositions of the present invention relates to novel lactic acid bacteria, their culture, their debris or probiotic activity in addition to its extracts, antimicrobial activity or beta-one kinds of the active ingredients a known having a galactosidase the (β-galactosidase) activity Or more. The pharmaceutical composition of the present invention can be formulated into a formulation for oral administration or parenteral administration by a conventional method, and can be formulated into a pharmaceutical composition such as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients. 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 ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Further, it can be suitably formulated according to each disease or ingredient, using appropriate methods in the art or by the method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA. The pharmaceutical composition of the present invention may be administered orally or parenterally to a mammal including a human according to a desired method. Examples of the parenteral administration method include external dermal application, intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, Intravenous injection or intra-thoracic injection. The dosage of the pharmaceutical composition of the present invention is not limited as long as it is a pharmacologically effective amount and is not limited as long as it depends on the body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, Varies. The typical daily dose of the pharmaceutical composition of the present invention is not particularly limited, but is preferably 0.1 to 3000 mg / kg, more preferably 1 to 2000 mg / kg, based on the active ingredient, once a day Or may be administered in divided doses.

The content of the novel lactic acid bacteria, the culture thereof, the lysate thereof or the extract thereof as an active ingredient in the food composition according to the present invention is preferably 0.01 to 99% by weight, more preferably 0.1 to 50% by weight, But is not limited thereto. The food composition of the present invention may be in the form of a pill, a powder, a granule, an infusion, a tablet, a capsule, or a liquid preparation. Examples of the food include meat, sausage, bread, chocolate, candy, snack, Other noodles, gums, dairy products including ice cream, various soups, drinks, tea, functional water, drinks, alcoholic beverages and vitamin complexes. In addition to the active ingredient, the food composition of the present invention may contain a pharmaceutically acceptable carrier, various flavors or natural carbohydrates as an additional ingredient. In addition, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The above-mentioned natural carbohydrates are sugar alcohols such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Natural flavors such as tau Martin and stevia extract, and synthetic flavors such as saccharin and aspartame may be used as the flavor.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following examples are intended to clearly illustrate the technical features of the present invention and do not limit the scope of protection of the present invention.

1. Isolation and Identification of Lactic Acid Bacteria

(1) Isolation of candidate lactic acid bacteria having probiotic activity

The candidate lactobacilli were screened from fermented cow milk curd. Fermented cow milk curd is a southern Indian traditional food that has been consumed safely by humans for at least 50 years. Screening of the candidate Lactobacillus was performed using a method known to those of ordinary skill in the art of biology and a selective MRS medium (Difco, USA) for lactic acid bacteria. The MRS medium supports the growth of lactic acid bacteria. Specifically, a diluted solution of fermented cow milk curd was plated on selective MRS medium for lactic acid bacteria and cultured at 37 ° C for about 24 hours. Then, the candidate strains that formed the colonies were separated, cultured on the medium, and then the strains were purified.

(2) Selection and identification of the final lactic acid bacteria

Probiotic lactic acid bacteria should inhibit the growth of Gram-negative bacteria such as E. coli and other enteric bacteria and may inhibit the growth of hemolytic activity, bioamine production, mucin degradation, should not be active in an in vitro safety assessment such as antibiotic susceptibility. Three final lactic acid bacteria were selected according to the above criteria, and the strain names of the three final lactic acid bacteria selected were designated as "MJM60319", "MJM60298" and "MJM60399", respectively. Figure 1 shows the morphology of the finally selected strains MJM60319, MJM60298 and MJM60399. Phylogenetic analysis was then performed on the three final lactic acid bacteria selected. First, the genomic DNA of the strain was isolated according to the manufacturer's instructions using a genomic DNA isolation kit (GeneALL, Seoul, Republic of Korea). PCR was then performed using forward primer 27F ((5'-AGAGTTTGATCCTGGCTCAG-3 ') and reverse primer 1492R (5'-GGTTACCT TGTTACGACTT-3') to amplify 16S rDNA of the strain. (5'-AGAGTTTGATCCTGGCTCAG-3 ') and 785F (5'-GGATTAGATACCCTGGTA (SEQ ID NO: 2)) as a primer were analyzed by using SolGent sequencing company (Republic of Korea) The 16S rDNA of the strain MJM60319 was found to have the contiguous base sequence of SEQ ID NO: 1, the 16S rDNA of the strain MJM60298 had the contiguous base sequence of SEQ ID NO: 2, and the 16S rDNA of the strain MJM60399 rDNA was found to have the contiguous nucleotide sequence of SEQ ID NO: 2. The 16S rDNA nucleotide sequences of the strains MJM60319, MJM60298 and MJM60399 were then compared to the GeneBank nucleotide database using the BLAST program and the MEGA 6 software (Tamura et al. To To complete the phylogenetic tree of strains MJM60319, MJM60298 and MJM60399.

Figure 2 is a diagram showing the phylogenetic tree of strains MJM60319, MJM60298 and MJM60399. As shown in FIG. 2, the 16S rDNA sequences of strains MJM60319, MJM60298 and MJM60399 showed 99% or more homology with 16S rDNA of Lactobacillus plantarum . Through this, strains of strains MJM60319, MJM60298 and MJM60399 were identified as Lactobacillus plantarum .

(3) Consignment information of lactic acid bacteria

The inventors of the present invention deposited a patent on Lactobacillus plantarum strain MJM60319 on December 1, 2015 to the National Institute of Agricultural Science and Technology, National Institute of Agricultural Science (National Institute of Agricultural Science and Technology, 370, Wonsan-gu, Jeonju, KACC 92111P. In addition, the inventors of the present invention have deposited a patent on Lactobacillus plantarum strain MJM60298 on December 1, 2015, at the National Institute of Agricultural Science and Technology (National Institute of Agricultural Science and Technology, National Institute of Agricultural Science and Technology, 370, Wonsan-gu, Jeonju, Korea) It was granted accession number KACC 92110P. In addition, the inventors of the present invention have deposited a patent on Lactobacillus plantarum strain MJM60399 on December 1, 2015, at the National Institute of Agricultural Science and Technology (National Institute of Agricultural Science and Technology, Agricultural Biotechnology Bank, Jeonju, Korea) I have been granted accession number KACC 92112P.

2. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , In vitro safety evaluation of MJM60298 and MJM60399

(1) Evaluation of biogenic amine formation

L-lysine, L-tryptophan, L-tyrosine, L-argnine, L-ornithine and L-histidine were added to the decarboxylase culture medium. Bover-Cid and Holzapfel (Bover-Cid S, Hozapfel WH.) Improved screening procedure for biogenic amine production by lactic acid bacteria. International Journal of Food Microbiology, 1999). Lactobacillus rhamnosus , a commercial probiotic strain, was used as a comparative strain, and Lactobacillus brevis having a bioamine production activity as a positive control strain was used. The composition of Bover-Cid and Holzapfel media is as follows.

Compostion  of Bover - Cid  and Holzapfel  medium

Tryptone: 0.5 wt%; yeast extract: 0.5 wt%; beef extract: 0.5 wt%; sodium chloride: 0.25 wt%; glucose: 0.05 wt%; tween 80: 0.1 wt%, MgSO 4 : 0.02 wt%; MnSO 4 : 0.005 wt%; FeSO4: 0.004 wt%; Ammonium citrate: 0.2 wt%; Thiamine: 0.001 wt%; dipotassium phosphate: 0.2 wt%; calcium carbonate: 0.01 wt%; pyridoxal 5-phosphate: 0.005 wt%; Amino acid (1 of the amino acid listed above): 1 wt%; bromocresol purple: 0.006 wt%; agar 2 wt%; pH: 5.3

FIG. 3 is a graph showing the results of measurement of the production of biologically-derived amines of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. The control photograph shown in Fig. 3 shows the results of experiments using a decarboxylase medium without amino acid. "MJM60319" term used in Figure 3 denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60319, "MJM60298" denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60298, "MJM60399" is Lactobacillus Flags Lactobacillus plantarum strain MJM60399. As shown in FIG. 3, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were found not to produce a biogenic amine.

(2) Evaluation of mucin degradation

Mucin degradation of the test strain was confirmed using agarose medium supplemented with 0.3% mucin. The medium was tryptone 7.5 g / l; casitone 7.5 g / l; yeast extract 3.0 g / l; meat extract 5.0 g / l; 5.0 g / l NaCl; 3.0 g / l K 2 HPO 4 .3H 2 O; 0.5 g / l KH 2 PO 4 ; MgSO 4 · 7H 2 O 0.5 g / ℓ; 0.5 g / l cysteine HCl; resazurin 0.002 g / l; D - (+) - glucose 3 g / l; 0.5 g / l of purified HGM and 1.5 g / l of agarose as a basic component. The pH of the medium was adjusted to about 7.2 using 2N NaOH aqueous solution. 10 [mu] L of the 24-hour culture medium of the test strain was inoculated on an agarose medium in Petri dish and incubated at 37 [deg.] C for 72 hours anaerobically. The medium was then stained with a colorant (0.1% Amido black in 3.5 M acetic acid) for 30 minutes and washed with 1.2 M acetic acid until a mucin lysis zone appeared around the colonies. The mucinolytic activity was defined as the size of the mucinolytic region. Lactobacillus rhamnosus , a commercial probiotic strain, was used as a comparative strain and a Bacillus sp. Strain having a mucinolytic activity as a positive control strain was used.

Fig. 4 is a diagram showing the results of measurement of mucin decomposition of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. "MJM60319" term used in Fig. 4 denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60319, "MJM60298" denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60298, "MJM60399" is Lactobacillus Flags Lactobacillus plantarum strain MJM60399. As shown in FIG. 4, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were confirmed to not undergo mucin digestion.

(3) Evaluation of hemolytic activity

Hemolytic activity of the test strains was assessed using a Columbia blood agar base (BD) containing 5% defibrinated sheep blood. When a clearance zone appears around the colony in the blood agar medium, it means that the hemolytic activity is positive, and when the clearance zone does not appear, the hemolytic activity is negative. Lactobacillus rhamnosus , a commercial probiotic strain, was used as a comparative strain in the evaluation of hemolytic activity, and Bacillus sp. Strain ID4 having hemolytic activity as a positive control strain was used.

FIG. 5 is a graph showing the results of measurement of hemolytic activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. "MJM60319" also the term used in 5 denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60319, "MJM60298" denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60298, "MJM60399" is Lactobacillus Flags Lactobacillus plantarum strain MJM60399. As shown in FIG. 5, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were found to show no hemolytic activity.

(4) Antimicrobial susceptibility evaluation

The susceptibility of the experimental strains Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 to antibiotics was determined by the inhibition of strain growth by various antibiotics. Several commercially available antibiotics were loaded on a filter paper disk of 8 mm diameter to a concentration of 100 μg per disc. The discs loaded with antibiotics were then placed on MRS agar plates each plated on the test strains and incubated at 37 ° C for 24-36 hrs. The larger the growth inhibition area of the test strain on the filter paper disk, the greater the susceptibility of the test strain to antibiotics. Table 1 below shows the results of the susceptibility test for the antibiotics of the experimental strains Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399.

Antibiotic action mechanism Antibiotic type Growth Inhibition Area of Experimental Strain MJM60319 MJM60298 MJM60399 Inhibition of protein synthesis Chloramphenicol 35 40 25 Tetracyclin 18 21 12 Lincomycin 25 50 25 Erythromycin 3.9 46 31 Streptomycin (NR) 1.5 16 R Inhibition of cell wall synthesis Penicillin 2.0 13 17 Ampicillin 47 50 35 Inhibition of bacterial DNA gyrase Novobiocin 24 35 17 Inhibition of bacterial DNA dependent protein synthesis Rifampicin 23 35 23

* NR: Antibiotic susceptibility is not required due to inherent resistance in EFSA, 2012; R: Resistant

The susceptibility of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 to antibiotics was assessed by the European Food Safety Authority (EFSA) recommendation [EFSA (2012) Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA Journal. 10: 2740.]. Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 are chloramphenicol, tetracycline, Lincomycin, Erythromycin, Streptomycin, Penicillin, Ampicillin, Novobiocin, and Lactobacillus plantarum according to the recommendations of the European Food Safety Authority (EFSA) Rifampicin, and other antibiotics.

3. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , Antimicrobial activity test of MJM60298 and MJM60399

(1) Experimental method

Antimicrobial activity tests of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were carried out using harmful microorganisms of human, animal and food. The culture broth of the test strain was filtered with cellulose acetate filimeter (pore size 0.2 탆), and the filtrate was concentrated 10 times using a centrifugal vacuum evaporator (Buchi, Switzerland). The concentrated culture broth was mixed with sterilized distilled water Lt; / RTI > Then, 100 占 퐇 of the diluted concentrated culture medium was loaded on a paper disk of 8 mm diameter and dried. Thereafter, the paper disk was placed on the LB agar plate coated with the harmful microorganism and incubated for 18 hours, and the inhibition region was confirmed. Lactobacillus rhamnosus , a commercial probiotic strain, was used as a comparative strain, and chloramphenicol, streptomycin and tetracycline were used as positive control drugs, respectively. The amount of positive control drug was 100 μg per paper disc. Specifically, chloramphenicol is an Escherichia coli O138 KCTC2615, Escherichia coli O1 KCTC2441, Escherichia coli Were applied to the growth inhibition test of K99 and Staphylococcus aureus KCCM11335, Streptomycin was applied to a growth inhibition test of Salmonella gallinarum and Salmonella typhimurium KCTC2931 KCTC2514, tetracycline is applied to the growth inhibition test of Salmonella choleraesuis KCTC2932. The experiment was repeated three times to check the reproducibility.

(2) Measurement results

6 is a diagram showing the results of measurement of antimicrobial activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. The results of measurement of antimicrobial activities of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 are shown in Table 2 below. As shown in FIG. 6 and Table 2, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were all Escherichia coli O138 KCTC2615, Escherichia coli O1 KCTC2441, Escherichia coli K99, Staphylococcus aureus KCCM11335, Salmonella gallinarum KCTC2931, inhibit the growth of harmful bacteria such as Salmonella typhimurium KCTC2514, Salmonella choleraesuis KCTC2932 effectively. From this, it can be seen that Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 produce antimicrobial substances which inhibit the growth of harmful bacteria.

Harmful microorganisms The growth inhibition area (cm) of the test strain culture solution or the test drug against the harmful microorganism MJM60319 MJM60298 MJM60399 L. rhamnosus Antibiotic E. coli O1 KCTC2441 1.6 1.8 1.9 1.1 2.1 (C) E. coli O138 KCTC2615 1.3 1.4 1.2 1.1 2.6 (C) E. coli K99 1.4 1.4 1.1 1.1 2.5 (C) Salmonella gallinarum KCTC2931 3.1 3.2 2.7 2.7 1.4 (S) Staphylococcus aureus KCCM11335 1.7 1.8 1.7 1.7 2.6 (C) Salmonella typhimurium KCTC2514 1.7 2.0 1.6 1.6 1.8 (S) Salmonella choleraesuis KCTC2932 1.7 2.1 1.6 1.6 1.5 (T)

C: chloramphenicol; S: Streptomycin; T: tetracycline

4. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 in oral-gastrointestinal transit conditions

(1) Experimental method

[Bove, P., Gallone, A., Russo, P., Capozzi, V., Albenzio, M., Spano, G., and Fiocco, D. 2012. Probiotic features of Lactobacillus plantarum mutant strains. Applied microbiology and biotechnology 96 (2): 431-441. Oro-gastrointestinal transit tolerance assay was performed by slightly modifying the method described in doi: 10.1007 / s00253-012-4031-2.

The test strain (1 × 10 9 / ml) was dissolved in an electrolyte solution (NaCl, 6.2 g / l; KCl, 2.2 g / l; CaCl 2 , 0.22g / ℓ; NaHCO 3, 1.2g / ℓ) were incubated for 10 minutes at [Marteau, P., Minekus, M. , Havenaar, R., and Huis in't Veild, JH 1997. Survival of lactic acid Cells cultured under oral stress conditions were centrifuged (1,800 × g, 5 min), and the cells were cultured under the conditions of oral stress. And then gastric stress was applied by incubating the cells in the stomach electrolyte solution containing 0.3% pepsin and pH 3 for 1 hour. After that, the cells were cultured under gastric stress conditions Cells were separated by centrifugation and incubated overnight at 37 ° C containing 0.1% pancreatin and 0.3% bile salts (even oxgall) Small intestine electrolyte solution (NaCl, 5g / ℓ; KCl , 0.6g / ℓ; CaCl 2, 0.25g / ℓ). Was incubated for 120 minutes at [. Marteau et al 1997] In addition, the test strains in PBS buffer, with no stress After the respective stresses were applied, the culture solution was sampled and the viability of the cells was evaluated by colony forming units (CFU) values. Specifically, the sampled cell culture medium was diluted with an appropriate amount of water to prepare a suspension The colony forming units (CFU) were calculated by incubating the suspension on an MRS agar medium and incubating for 48 hours at 37 ° C. The number of viable strains surviving each stress treatment was calculated as Log 10 CFU 10 of colony forming units).

(2) Measurement results

FIG. 7 is a graph showing the results of resistance evaluation of Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 under oral-gastrointestinal transit conditions. "MJM60319" also the term used in 7 denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60319, "MJM60298" denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60298, "MJM60399" is Lactobacillus Flags Lactobacillus plantarum strain MJM60399. In addition, the survival percentages of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 were shown in Table 3 when the Oro-gastrointestinal transit tolerance assay (OGT assay) was completed. As shown in FIG. 7 and Table 3, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 showed high resistance to oral-gastrointestinal transit conditions. Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 are estimated to have a high survival rate, considering that most of the lactic acid bacteria are not viable and die in oral-gastrointestinal transit conditions. In addition, survival rates in oral-gastrointestinal transit conditions of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 can be further improved by appropriate protective agents or microencapsulation.

Test strain Survival cells after OGT assay% MJM60319 5.7 MJM60298 6.1 MJM60399 6.9

5. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 on human epithelial-like Caco-2 cell lines Attachment  evaluation

(1) Experimental method

The Caco-2 cell line was distributed from the microbial resource center (KCTC) and used for the epithelial cell adhesion test. Experiments were performed according to the method described in [Tuo et al., 2013].

First, Caco-2 cells were added to MEM (Minimum Essential Media) medium (Gibco, USA) supplemented with 20% (v / v) inactivated fetal bovine serum (FBS) and 1 unit of penicillin-streptomycin antibiotic Incubated for 21 days in an incubator with a temperature of 37 < 0 > C and a CO 2 concentration environment of 5%.

Subsequently, well-differentiated Caco-2 cells were plated in 12-well plates at a concentration of 5 × 10 2 cells / ml, cultured for 3 days while exchanging the medium daily, and a monolayer filled with 80% / RTI > Caco-2 cells in a monolayer were reacted with 0.25% trypsin-EDTA (Gibco, USA) solution at 37 ° C for 5 minutes to obtain single cells. The cells were injected into a hemocytometer, Respectively.

Next, single layer Caco-2 cells were cultured in MEM medium without antibiotics for 2 hours. The culture broth was mixed with the FBS solution containing 0.5 ml of the test strain at the same volume to give a final bacterial concentration of 1 × 10 6 CFU / ml. Then, the mixed solution was inoculated into a 12-well plate (Well Plates) and cultured in a 5% CO 2 environment at 37 ° C for 2 hours. Thereafter, the single cell layer was washed three times with sterilized PBS (pH 7.2) solution to remove unattached test strains. Thereafter, 1 ml of 0.05% (v / v) Triton-X100 was added to the wells to separate the attached test strains and Caco-2 cells, and the colony forming units (CFU) of the attached test strains were calculated Adhesion ability was evaluated. The percentage of adhesion of the test strain to the Caco-2 cell line was calculated using the following formula.

Figure 112016012946261-pat00001

(2) Measurement results

8 is a photograph showing the adherence ability of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 to Caco-2 cell monolayer. 8, the scale bar indicates 10 mu m.

As shown in FIG. 8, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 showed excellent adhesion to intestinal epithelial cells. These results indicate that the novel strains according to the present invention are excellent in adhesion to intestinal epithelial cells It was confirmed that the intestinal environment can be improved.

6. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399

(1) Experimental method

[Guo, C. F., Zhang, L. W., Han, X., Li, J. Y., Du, M., Yi, H. X., Feng, Z., Zhang, Y. C., and Xu, X.R. 2011. A sensitive method for qualitative screening of even salt hydrolase-active lactobacilli based on thin-layer chromatography. Journal of dairy science 94 (4): 1732-1737. doi: 10.3168 / jds.2010-3801] was slightly modified to qualitatively measure the bile salt hydrolase activity of the test strains.

First, MRS reaction liquid medium was prepared by using sodium phosphate buffer (pH 7.4) containing 5 mM taurodeoxycholic acid (TDCA), glycocholic acid (GCA) and taurocholic acid (TCA) In addition, the test strain was cultured overnight, washed twice with PBS solution (pH 7.2), and suspended in PBS solution (pH 7.2) to prepare a test strain suspension. Thereafter, the test strain suspension was mixed with an equal volume of MRS reaction liquid medium and incubated at 37 DEG C for 16 hours. Thereafter, the mixed culture was lyophilized and dissolved in 1 ml of methanol. 3 μl of the methanol suspension was spotted on a silica gel 60 (20 cm × 20 cm), and a mobile phase (composed of a solvent system having hexane: methylethlyketone: glacial acetic acid having a volume ratio of 56: 36: 8) [Chavez, MN, and Krone, CL 1976. Silicic acid thin-layer chromatography of conjugated and free bile acids. Journal of lipid research 17 (5): 545-547]. Thereafter, the TLC plate was dried, sprayed with 10% aqueous sulfuric acid solution, heated at 110 ° C. for 10 minutes, and free cholic acid, deoxycholic acid, and conjugated bile acid Respectively.

(2) Measurement results

The results of the measurement of the activity of the bile acid hydrolytic enzymes of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 are shown in Table 4 below. As shown in the following Table 4, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 showed bile salt hydrolase activity, and TDCA, TCA and GCA were decompounded to form free cholic acid or de And converted to deoxycholic acid.

Test strain TDCA hydrolytic activity TCA hydrolytic activity GCA hydrolytic activity MJM60319 + + + MJM60298 + + + MJM60399 + + +

7. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 beta-galactosidase ( β-galactosidase ) Active measurement

The test strain was inoculated on MRS agar medium containing X-Gal (20 mg / ml in DMSO) on the surface and cultured at 37 DEG C for 16 hours. The presence of blue color in the colonies of the test strain indicates the presence of beta-galactosidase activity.

9 is a photograph showing the results of measurement of beta -galactosidase activity of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. "MJM60319" term used in Figure 9 denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60319, "MJM60298" denotes a Lactobacillus Planta column (Lactobacillus plantarum) strain MJM60298, "MJM60399" is Lactobacillus Flags Lactobacillus plantarum strain MJM60399. As shown in FIG. 9, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 showed beta-galactosidase activity. Thus, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 can be used to alleviate lactose intolerance in pets such as cats.

8. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 < RTI ID = 0.0 >

(1) Experimental method

Was inoculated into an MRS liquid medium containing sodium chloride at a concentration of 1 to 10% by weight and inoculated with 1 ml of the test strain culture medium (OD 0.5) and cultured at 37 캜 for 24 hours. In addition, the test strain culture solution was inoculated to the MRS liquid medium containing no sodium chloride and cultured, and then used as a control. After incubation, the absorbance of the culture was measured at a wavelength of 600 nm using a 96-well plate reader (Infinite M200® PRO, Tecan Austria GmbH, Untersbergstr). Growth of the test strain was expressed as% relative growth based on the control group and calculated using the following equation.

Figure 112016012946261-pat00002

Ac is the absorbance value of the control group, and At is the absorbance value of the sample cultured in the MRS liquid medium with various sodium chloride concentrations.

(2) Measurement results

10 is a graph showing the growth levels of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 under various sodium chloride concentrations. "MJM60319" refers to Lactobacillus plantarum strain MJM60319, "MJM60298" refers to Lactobacillus plantarum strain MJM60298, "MJM60399" refers to Lactobacillus plantarum strain, Lactobacillus plantarum strain MJM60399. As shown in FIG. 10, all of the Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 showed a growth pattern that decreased with increasing sodium chloride concentration. Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 can be estimated to have excellent sodium chloride tolerance with a survival rate of 60% or more even at a sodium chloride concentration of 8%. Sodium chloride tolerance is one of the important technical features of strains because it helps to maintain stability and viability under conditions for producing bacteria for probiotics.

9. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 to evaluate the ability to produce D, L-lactate

The test strain was inoculated into the MRS liquid medium and cultured at 37 DEG C for 24 hours. Then, the supernatant was collected from the culture of the test strain, and the amounts of D-lactate and L-lactate were analyzed using the D, L-lactate assay kit (Megazyme, Ireland, Cat. No. K-DLATE).

11 is a graph showing the production ability of D, L-lactate of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399. 11, "MJM60319" represents Lactobacillus plantarum strain MJM60319, "MJM60298" represents Lactobacillus plantarum strain MJM60298, "MJM60399" represents Lactobacillus plantarum strain, Lactobacillus plantarum strain MJM60399. As shown in FIG. 11, Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 produced both D-lactate and L-lactate and produced more D-lactate than L-lactate. However, the differences in the yields of D-lactate and L-lactate are within acceptable ranges.

10. Lactobacillus Planta Rum  ( Lactobacillus plantarum ) Strain MJM60319 , MJM60298 and MJM60399 in freeze-drying conditions

The survival rate of the initial test strain of the test strain culture was adjusted to 1 x 10 10 CFU / ml and 5% (w / v) skim milk powder was added to the cryoprotectant, After adding 5% (w / v) of maltodextrin powder, it was frozen at -80 ° C and lyophilized for 18 hours under reduced pressure (5 mbar). After lyophilization, the sample was rehydrated with sterilized water. Thereafter, the CFU of the test strain was measured, and the survival number of the test strain was calculated as a relative survival value in comparison with the survival count of the initial test strain.

12 is a graph showing the results of measurement of the survival stability of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 under lyophilization conditions. "MJM60319" refers to Lactobacillus plantarum strain MJM60319, "MJM60298" refers to Lactobacillus plantarum strain MJM60298, "MJM60399" refers to Lactobacillus plantarum strain, Lactobacillus plantarum strain MJM60399, and "CP" represents a cryoprotectant. 12, when 5% (w / v) of skim milk powder and 5% (w / v) of maltodextrin powder were added as a cryoprotectant, Lactobacillus sp. The survival stability of Lactobacillus plantarum strains MJM60319, MJM60298 and MJM60399 was greatly improved.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. Therefore, the scope of the present invention should be construed as including all embodiments falling within the scope of the appended claims.

<110> Myongji University Industry and Academia Cooperation Foundation <120> Novel Lactobacillus plantarum with probiotic activities and use          the <130> DP-15-1024 <160> 3 <170> KoPatentin 3.0 <210> 1 <211> 1490 <212> DNA <213> Unknown <220> <223> 16S rDNA of Lactobacillus plantarum MJM60319 <400> 1 gggtgctata atgcaagtcg aacgaactct ggtattgatt ggtgcttgca tcatgattta 60 catttgagtg agtggcgaac tggtgagtaa cacgtgggaa acctgcccag aagcggggga 120 taacacctgg aaacagatgc taataccgca taacaacttg gaccgcatgg tccgagtttg 180 aaagatggct tcggctatca cttttggatg gtcccgcggc gtattagcta gatggtgggg 240 taacggctca ccatggcaat gatacgtagc cgacctgaga gggtaatcgg ccacattggg 300 actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc acaatggacg 360 aaagtctgat ggagcaacgc cgcgtgagtg aagaagggtt tcggctcgta aaactctgtt 420 gttaaagaag aacatatctg agagtaactg ttcaggtatt gacggtattt aaccagaaag 480 ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggat 540 ttattgggcg taaagcgagc gcaggcggtt ttttaagtct gatgtgaaag ccttcggctc 600 aaccgaagaa gtgcatggga aactgggaaa cttgagtgca gaagaggaca gtggaactcc 660 atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag gcggctgtct 720 ggtctgtaac tgacgctgag gctcgaaagt atgggtagca aacaggatta gataccctgg 780 tagtccatac cgtaaacgat gaatgctaag tgttggaggg tttccgccct tcagtgctgc 840 agctaacgca ttaagcattc cgcctgggga gtacggccgc aaggctgaaa ctcaaggaat 900 tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagctac gcgaagaacc 960 ttaccaggtc ttgacatact atgcaaatct aagagattag acgttccctt cggggacatg 1020 gatacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc 1080 aacgagcgca acccttatta tcagttgcca gcattaagtt gggcactctg gtgagactgc 1140 cggtgacaaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg 1200 gctacacacg tgctacaatg gatggtacaa cgagttgcga actcgcgaga gtaagctaat 1260 ctcttaaagc cattctcagt tcggattgta ggctgcaact cgcctacatg aagtcggaat 1320 cgctagtaat cgcggatcag catgccgcgg tgaatacgtt cccgggcctt gtacacaccg 1380 cccgtcacac catgagagtt tgtaacaccc aaagtcggtg gggtaacctt ttaggaacca 1440 gccgcctaag gtgggacaga tgattagggt gaagtcgtaa caagtaaacc 1490 <210> 2 <211> 1486 <212> DNA <213> Unknown <220> <223> 16S rDNA of Lactobacillus plantarum MJM60298 <400> 2 gctataatgc aagtcgaacg aactctggta ttgattggtg cttgcatcat gatttacatt 60 tgagtgagtg gcgaactggt gagtaacacg tgggaaacct gcccagaagc gggggataac 120 acctggaaac agatgctaat accgcataac aacttggacc gcatggtccg agtttgaaag 180 atggcttcgg ctatcacttt tggatggtcc cgcggcgtat tagctagatg gtggggtaac 240 ggctcaccat ggcaatgata cgtagccgac ctgagagggt aatcggccac attgggactg 300 agacacggcc caaactccta cgggaggcag cagtagggaa tcttccacaa tggacgaaag 360 tctgatggag caacgccgcg tgagtgaaga agggtttcgg ctcgtaaaac tctgttgtta 420 aagaagaaca tatctgagag taactgttca ggtattgacg gtatttaacc agaaagccac 480 ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt ccggatttat 540 tgggcgtaaa gcgagcgcag gcggtttttt aagtctgatg tgaaagcctt cggctcaacc 600 gaagaagtgc atcggaaact gggaaacttg agtgcagaag aggacagtgg aactccatgt 660 gtaggggga aatgcgtaga tatatggaag aacaccagtg gcgaaggcgg ctgtctggtc 720 tgtaactgac gctgaggctc gaaagtatgg gtagcaaaca ggattagata ccctggtagt 780 ccataccgta aacgatgaat gctaagtgtt ggagggtttc cgcccttcag tgctgcagct 840 aacgcattaa gcattccgcc tggggagtac ggccgcaagg ctgaaactca aaggaattga 900 cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagctacgcg aagaacctta 960 ccaggtcttg acatactatg caaatctaag agattagacg ttcccttcgg ggacatggat 1020 acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080 gagcgcaacc cttattatca gttgccagca ttaagttggg cactctggtg agactgccgg 1140 tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat gacctgggct 1200 acacacgtgc tacaatggat ggtacaacga gttgcgaact cgcgagagta agctaatctc 1260 ttaaagccat tctcagttcg gattgtaggc tgcaactcgc ctacatgaag tcggaatcgc 1320 tagtaatcgc ggatcagcat gccgcggtga atacgttccc gggccttgta cacaccgccc 1380 gtcacaccat gagagtttgt aacacccaaa gtcggtgggg taacctttta ggaaccagcc 1440 gcctaaggtg ggacagatga ttagggtgaa gtcgtaacaa gttaaa 1486 <210> 3 <211> 1467 <212> DNA <213> Unknown <220> <223> 16S rDNA of Lactobacillus plantarum MJM60399 <400> 3 ggcggcgtgc tatacatgca agtcgaacga actctggtat tgattggtgc ttgcatcatg 60 atttacattt gagtgagtgg cgaactggtg agtaacacgt gggaaacctg cccagaagcg 120 ggggataaca cctggaaaca gatgctaata ccgcataaca acttggaccg catggtccga 180 gtttgaaaga tggcttcggc tatcactttt ggatggtccc gcggcgtatt agctagatgg 240 tggggtaacg gctcaccatg gcaatgatac gtagccgacc tgagagggta atcggccaca 300 ttgggactga gacacggccc aaactcctac gggaggcagc agtagggaat cttccacaat 360 ggacgaaagt ctgatggagc aacgccgcgt gagtgaagaa gggtttcggc tcgtaaaact 420 ctgttgttaa agaagaacat atctgagagt aactgttcag gtattgacgg tatttaacca 480 gaaagccacg gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc 540 cggatttatt gggcgtaaag cgagcgcagg cggtttttta agtctgatgt gaaagccttc 600 ggctcaaccg aagaagtgca tcggaaactg ggaaacttga gtgcagaaga ggacagtgga 660 actccatgtg tagcggtgaa atgcgtagat atatggaaga acaccagtgg cgaaggcggc 720 tgtctggtct gtaactgacg ctgaggctcg aaagtatggg tagcaaacag gattagatac 780 cctggtagtc cataccgtaa acgatgaatg ctaagtgttg gagggtttcc gcccttcagt 840 gctgcagcta acgcattaag cattccgcct ggggagtacg gccgcaaggc tgaaactcaa 900 aggaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agctacgcga 960 agaaccttac caggtcttga catactatgc aaatctaaga gattagacgt tcccttcggg 1020 gacatggata caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag 1080 tcccgcaacg agcgcaaccc ttattatcag ttgccagcat taagttgggc actctggtga 1140 gactgccggt gacaaaccgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg 1200 acctgggcta cacacgtgct acaatggatg gtacaacgag ttgcgaactc gcgagagtaa 1260 gctaatctct taaagccatt ctcagttcgg attgtaggct gcaactcgcc tacatgaagt 1320 cggaatcgct agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac 1380 acaccgcccg tcacaccatg agagtttgta acacccaaag tcggtggggt aaccttttag 1440 gaaccagccg cctaaggtgg gacagat 1467

Claims (16)

Lactobacillus plantarum MJM60319 (accession number: KACC 92111P).
delete A probiotic composition comprising Lactobacillus plantarum MJM60319 (Accession No: KACC 92111P) strain or a culture thereof as an active ingredient.
The probiotic composition according to claim 3, further comprising a skim milk powder or a maltodextrin powder.
One or more strains selected from the group consisting of Lactobacillus plantarum MJM60298 (accession number: KACC 92110P) or Lactobacillus plantarum MJM60399 (accession number: KACC 92112P) and Lactobacillus plantarum ( Lactobacillus plantarum ) MJM60319 (accession number: KACC 92111P) or a culture of the mixed lactic acid bacterium as an active ingredient.
delete delete The probiotic composition according to claim 5, further comprising a skim milk powder or a maltodextrin powder.
A composition for antibacterial use comprising Lactobacillus plantarum MJM60319 (Accession No .: KACC 92111P) strain, a culture thereof, a lysate thereof or an extract thereof as an active ingredient.
The antimicrobial composition according to claim 9, further comprising a skim milk powder or a maltodextrin powder.
One or more strains selected from the group consisting of Lactobacillus plantarum MJM60298 (accession number: KACC 92110P) or Lactobacillus plantarum MJM60399 (accession number: KACC 92112P) and Lactobacillus plantarum ( Lactobacillus plantarum ) MJM60319 (accession number: KACC 92111P), a culture of the mixed lactic acid bacterium, a lysate of the mixed lactic acid bacteria or an extract of the mixed lactic acid bacterium as an active ingredient.
delete delete The antimicrobial composition according to claim 11, further comprising a skim milk powder or a maltodextrin powder.
A composition for preventing or ameliorating lactose intolerance comprising Lactobacillus plantarum MJM60319 (Accession No: KACC 92111P) strain, a culture thereof, a lysate thereof or an extract thereof as an active ingredient.
One or more strains selected from the group consisting of Lactobacillus plantarum MJM60298 (accession number: KACC 92110P) or Lactobacillus plantarum MJM60399 (accession number: KACC 92112P) and Lactobacillus plantarum Lactose intolerance prevention or prevention of lactose intolerance comprising the mixed lactic acid bacteria comprising Lactobacillus plantarum MJM60319 (Accession No: KACC 92111P), the culture of the mixed lactic acid bacteria, the lysate of the mixed lactic acid bacteria or the extract of the mixed lactic acid bacteria as an active ingredient, / RTI &gt;
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