KR20160057855A - Lactic acid bacterium separated from kimchii and having antifungal activity, and compositon including it - Google Patents

Abstract

The present invention relates to a novel strain of lactic acid bacteria separated from Kimchi and a composition comprising the same and, more specifically, to Lactobacillus sakei DSR KF14 having antifungal and antimicrobial activities, to a composition comprising the same, and to antimicrobial uses of the composition. According to the present invention, Lactobacillus sakei DSR KF14 of lactic acid bacteria has antimicrobial activities and antifungal activities, and can be efficiently used for sterilizing, antimicrobial, and antiseptic purposes in various fields such as a food, a feed, a drug, a cosmetic product, etc.

Description

TECHNICAL FIELD [0001] The present invention relates to a lactic acid bacterium having an antifungal activity isolated from kimchi and a composition containing the same. [0002]

The present invention relates to a novel lactic acid bacteria strain isolated from kimchi and a composition containing the same, and more particularly, to a lactic acid bacteria strain having an antibacterial activity and an antibacterial activity, such as Lactobacillus < / RTI > sakei ) DSR KF14 and compositions comprising same, and to the use of such compositions for antimicrobial use.

Preservatives such as food, cosmetics and medicines are additives used to prevent deterioration, deterioration and chemical change of foods, cosmetics and pharmaceuticals and to maintain nutritional value, freshness and quality. They are used to inhibit the growth of microorganisms such as bacteria, fungi and yeast, And additives used to preserve cosmetics and medicines.

Conventional preservatives or preservatives are mainly used as chemical preservatives and preservatives. In the case of children, the daily allowable dose can be easily exceeded, the possibility of inducing diseases such as asthma is caused, and the rapid and chronic toxicity And the possibility of new problems such as mutation induction are emerging.

In recent years, consumers are increasingly interested in health and have a sense of rejection of chemical synthetic food preservatives due to the above problems, and are aiming to be fresher, natural as well as no chemical preservatives. With this demand, there is a growing need for studies on natural preservatives that are safe and economical.

Up to now, mainly synthetic or natural antibiotics have been used to prevent or treat infections caused by such harmful microorganisms. However, the use of synthetic antibiotics is strictly regulated because of its resistance and residuals, and it also destroys intestinal beneficial microflora, thus losing the preventive effect against pathogenic bacteria infection by normal microflora. Although alkaloids, flavonoids, phytoalexins, antimicrobial peptides, organic acids and fatty acids are known as natural antibiotics, most of them are not commercialized due to problems of color, stability, narrow antibacterial spectrum and shape In fact.

With regard to the development of natural preservatives and substances having antifungal activity, there is growing interest in antibiotics produced from microorganisms or microorganisms. Lactobacillus is generally regarded as safe because it has been widely used in food, both east and west, and is generally considered safe.

As antibacterial substances produced by lactic acid bacteria, organic acids such as lactic acid, acetic acid, hydrogen peroxide, bacteriocin and the like have been known. Bacteriocin has the advantage of being safe, but its antibacterial spectrum is so narrow that it has not been widely used as a food preservative because of its disadvantage that it has almost no antimicrobial activity against Gram-negative bacteria, yeast and fungi.

There are several kinds of lactic acid bacteria in Kimchi, and there are reports of antibacterial substances in these lactic acid bacteria. However, there are very few reports on lactic acid bacteria having antifungal activity among kimchi - derived lactic acid bacteria. There is an urgent need to search for microorganisms capable of producing excellent antifungal substances capable of solving the limitations of the substances studied so far and to study the substances produced by the microorganisms.

Korean Patent Registration No. 10-1124056 (published on March 23, 2012) Korea Patent Office Registration No. 10-1381547 (Announcement on Mar. 28, 2014) Patent Registration No. 10-1256727 (published on April 15, 2013)

The inventors of the present invention have found that lactic acid bacteria have antimicrobial activity only on ordinary bacteria while continuing their research to solve the problems of the prior art as described above, Can be separated from the kimchi. Thus, the present invention has been completed.

Accordingly, an object of the present invention is to provide a novel lactic acid bacteria having antifungal and antibacterial activity.

Another object of the present invention is to provide antifungal and antibacterial compositions comprising the antifungal and antibacterial activity of lactic acid bacteria.

Yet another object of the present invention is to provide foods, feeds, medicines and cosmetics containing the antifungal and antibacterial compositions.

To achieve these and other advantages and in accordance with the purpose of the present invention, as embodied and broadly described herein, there is provided a strain of Lactobacillus saccharum DSR KF14 having an antifungal activity and an antibacterial activity.

The present invention also provides antifungal and antibacterial compositions comprising the Lactobacillus saccharum DSR KF14 strain.

The present invention also provides foods, feeds, medicines, and cosmetics comprising the antifungal and antibacterial compositions.

The lactobacillus case DSR KF14 of the present invention is a lactic acid bacterial strain isolated from kimchi and has antifungal activity and antimicrobial activity, and is excellent in acid resistance and bile resistance.

In particular, the lactic acid bacterium of the present invention exhibits antibacterial activity against fungi such as molds and yeast as well as bacteria, and Lactobacillus casei DSR KF14, the culture liquid thereof and the fungal substance are added to foods, feeds, medicines and cosmetics, It can also be developed as a treatment for fungal diseases.

In addition, when the kimchi is immersed using the lactic acid bacteria of the present invention, propagation of pathogenic microorganisms capable of early growth can be suppressed and a flavor of Kimchi can be obtained by lactobacillus sake DSR KF14 fermentation.

One aspect of the present invention relates to a strain of lactic acid Lactobacillus saccharum DSR KF14.

The lactobacillus case DSR KF14 of the present invention can be used as a strain itself or in the form of a culture throughout the invention. The lactobacillus case DSR KF14 of the present invention is safe even when the live bacteria are ingested by a vegetable lactic acid bacterium isolated from kimchi and has an effect of inhibiting the growth of food pathogens and pathogen-inducing yeasts by microorganisms.

That is, the strain according to the present invention is characterized by having an antibacterial activity and an antibacterial activity.

Examples of the fungi include fungi such as yeasts such as Candida spp., Zygosaccharomyces spp., Aspergillus spp., Penicillium spp., Fusarium spp. But is not limited thereto.

Examples of the bacteria include Bacillus sp., Staphylococcus sp., Listeria sp., Salmonella sp., Streptococcus, and the like. It is not.

According to an embodiment of the present invention, the Lactobacillus casei DSR KF14 strain is characterized by having acid resistance and biliary cholesterol.

The acid resistance may mean that the growth is active even at an acidity ranging from pH 1.0 to pH 3.5, preferably pH 2.0 to pH 3.0.

The intrinsic bile resistance may mean that the growth is active at an oxgall concentration of 0.1 to 5 wt%, and preferably 0.3 to 3 wt% of Oggal concentration.

In particular, the strain of the present invention has a strong acid tolerance and survival rate of 90% or more at a pH of 2.5, exhibiting a high survival rate of 100% at 3% by weight of oxgall, and exhibiting strong biteability.

Since the strain of the present invention is excellent in acid resistance and biliary excretion, when the strain is orally ingested, the acid environment in which the pH is lowered to 3 or less, and the digestive enzymes and bile acids secreted from the stomach and small intestine, The probability of reaching a good resolution can be increased.

The strain according to the present invention may be a vegetable lactic acid bacteria isolated from kimchi, but the source of the isolate is not limited thereto.

Another aspect of the present invention relates to an antifungal and antibacterial composition comprising the Lactobacillus saccharum DSR KF14 strain.

Since the Lactobacillus casei DSR KF14 strain has excellent antifungal activity and antibacterial activity, the lactic acid bacterium of the present invention can be effectively used for antibacterial use.

That is, the antifungal and antimicrobial composition of the present invention may be added to foods, feeds, medicines, cosmetics, and the like, but the antifungal and antibacterial composition may be used without limitation as long as the strain of the present invention can be included.

For example, when the composition is added to foods, the growth of harmful microorganisms can be effectively inhibited, the freshness of the food can be maintained, and the storage period and the circulation period can be effectively extended.

The composition may contain, as an active ingredient, a crushed cell wall fraction, a live cell, a dead cell, a dried bacterium or a culture of the lactic acid bacteria of the present invention, and may further contain an excipient or a carrier. The cultivation of the above-mentioned lactic acid bacteria can be carried out according to a suitable culture medium and culture conditions known in the art. Such a culturing process can be easily adjusted according to the strain selected by those skilled in the art. These various methods of culturing are disclosed in various publications (e.g., James et al., Biochemical Engineering, Prentice-Hall International Editions). Suspension cultures and adherent cultures can be divided into batches, fed-batch and continuous-batch cultures according to the culture method. The culture includes a culture medium itself cultured in a liquid medium, a filtrate (centrifuged supernatant) obtained by removing the strain by filtration or centrifugation of the culture medium, and the like. The content of the lactic acid bacteria in the composition may vary depending on the use of the composition and the formulation.

The antifungal and antimicrobial composition refers to a composition having an activity of inhibiting or suppressing the growth of microorganisms or directly killing microorganisms. The microorganisms include, but are not limited to, B. cereus, Staphylococcus aureus Staphylococcus aureus, Listeria monocytogenes, E. coli, Salmonella typimurium, Streptococcus mutans, and the like.

In addition, the composition of the present invention exhibits excellent antimicrobial activity against the transformed yeast and pathogenic fungi isolated from the over-cooked food, and can inhibit abnormal fermentation. Specifically, the yeast can be selected from the group consisting of Candida albicans, Zygosaccharomyces rouxii, and the fungus may be Aspergillus flavus, Penicillium expensum, or Fusarium oxysproum.

The composition of the present invention can be added to foods to be used as natural antimicrobial agents or preservatives. The term " food " means an entire food such as a health functional food, a functional food, a natural food, a processed food, a general food, and particularly when used in fermented foods such as kimchi and dairy products, . ≪ / RTI >

In addition, the lactic acid bacteria of the present invention can be used as a starter for the production of fermented foods.

The fermented food includes cheese, kimchi, fermented reproductive products and the like. The fermented food using the lactic acid bacterium of the present invention can be produced according to a conventional method known in the art. For example, lactic acid bacteria or a culture thereof according to the present invention is treated with a starter to ferment a cereal powder such as brown rice and yulmu at a proper temperature, and then various agricultural products such as white rice, glutinous rice and sorghum are suitably To produce a fermented reproductive product.

In particular, the lactic acid bacteria according to the present invention can be used for producing kimchi. Preferably, the kimchi can be prepared by adding a common kimchi seasoning such as red pepper powder, garlic, ginger, green onion, mungbean and sugar to the salted Chinese cabbage, and then adding a culture of the lactic acid bacteria of the present invention. The lactic acid bacterium culture of the present invention is preferably added in an amount of 0.1 to 5 wt% based on the total weight of the kimchi, and most preferably 1 wt%.

In addition, the composition of the present invention can be manufactured and used in the form of fermented feed, compound feed, pellet, and silage. The fermented feed can be prepared by fermenting an organic material by adding the lactic acid bacteria of the present invention and various microorganisms or enzymes, and the composition feed can be prepared by mixing various kinds of general diets and the lactic acid bacteria of the present invention. The pellet-shaped feed can be produced by formulating the fermented feed or the compounded feed into a pelletizer, and the silage can be produced by fermenting the fermented feed with the lactic acid bacteria according to the present invention.

In addition, the composition of the present invention may be a pharmaceutical composition capable of treating or preventing an infectious disease of harmful microorganisms, and may be manufactured and administered in various formulations and methods. For example, the lactic acid bacteria or culture thereof of the present invention may be mixed with a carrier commonly used in the pharmaceutical field to prepare tablets, troche, capsules, elixir, syrup, ), Powders, suspensions or granules, and 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 dose may be appropriately selected depending on the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, age, sex, condition, etc. of the subject.

In addition, the composition of the present invention can be manufactured and used in the form of usual cosmetics such as lotion, cream, gel and the like.

Hereinafter, the present invention will be described in detail with reference to Examples.

However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited by the following examples.

< Example  1> Isolation of Antimicrobial and Antifungal Activity Strain from Kimchi

<1-1> Isolation of lactic acid bacteria from kimchi

Various kinds of kimchi prepared by conventional kimchi manufacturing method were used as lactic acid bacteria isolated kimchi samples. Samples of the kimchi were collected and diluted 10 times with 0.85% saline, and each of them was diluted 10 times with 0.85 ml of saline, and 10 ml of MRS agar medium (MRS agar, Difco .; 10 g of bovine peptone, 10 g of beef extract, 5 g of yeast extract, , 2 g of citric acid, 2 g of dibasic potassium phosphate, 5 g of sodium acetate, 0.1 g of manganese sulfate, 0.05 g of magnesium sulfate and 15 g of agar / 1 l of distilled water). Plates were then incubated for 1 day in a 37 &lt; 0 &gt; C incubator. Each of the resulting colonies was inoculated on MRS agar plates and cultured at 37 ° C for 1 day to isolate lactic acid bacteria colonies.

<1-2> Screening of lactic acid bacteria having antibacterial and antifungal activity

a. Culture of strain

The lactic acid bacteria colonies isolated in Example <1-1> were cultured in an MRS liquid medium at 37 ° C. for 24 hours, and then the culture medium for the third culture was centrifuged, suspended in 10% skim milk, and stored at -20 ° C. Respectively.

Among the 2000 kinds of lactic acid bacteria isolated in Example <1-1>, in order to select lactic acid bacteria that inhibits the growth of harmful microorganisms, three kinds of harmful microorganisms, Bacillus cereus KCCM 12142 and Staphylococcus aureus, Staphylococcus aureus KCCM11335 and Escherichia coli KCCM11234 were inoculated into a nutrient medium (5.0 g of bacto peptone, 1.0 g of beef extract, 2.0 g of yeast extract, 5.0 g of sodium chloride, 15 g of agar / 1 l of distilled water) And cultured at 37 DEG C for 24 hours. The cultured cells were collected from the medium, washed twice with phosphate buffer (PBS, pH 7.4), suspended in 20% glycerol, and stored at -20 ° C.

Candida albicans KCTC17485 was cultured for 2 days at 25 ° C to 30 ° C using YPD (Yeast Peptone Dextrose) liquid medium (Difco Laboratories, USA). The cultured medium was centrifuged, suspended in 20% glycerol, and stored at -20 ° C.

Aspergillus flavus KCTC16682 was cultured in a PDA medium (potato dextrose agar, Difco Laboratories, USA) at 25 ° C to 30 ° C for 3 to 5 days.

Spores of the fungi were recovered to carry out experiments to confirm antifungal activity. The fungi were inoculated on a PDA medium and cultured at 25 ° C to 30 ° C for 3 to 5 days until spores were produced. The resulting spores were extracted with sterile water containing Tween 80 (Polyoxyethyelene Sorbitan Monooleate) at 0.05% (v / v). Extracted spores were stored at -80 ° C using a glycerol / water (20:80, v / v) solution.

b. Isolation of lactic acid bacteria having antibacterial and antifungal activity

The harmful microorganisms cultured in the third subculture on the nutrient liquid medium were inoculated 1% in the nutrient agar medium, hardened on the plate, and dried for 20 minutes. In the YPD liquid medium, the third strain of transformed yeast was inoculated 1% in YPD agar medium, hardened on the plate, and dried for 20 minutes.

The spores extracted above were inoculated into the PDA medium by 1%, hardened on the plate, and dried for 20 minutes. Thereafter, a hole having a diameter of 9 mm was drilled in a cockbearer, and 100 쨉 l of each of the separated lactic acid bacteria culture supernatant was dripped. The diameter of the inhibitory rings produced after incubation at 37 ° C for 24 hours (yeast at 25-30 ° C for 48 hours, fungus at 25-30 ° C for at least 48 hours) was measured. The strain LAB1 which selected the most excellent strains among the lactic acid bacteria forming the inhibitory ring and finally suppressed the growth of the harmful microorganisms was selected (Table 1).

Size of microbial growth inhibition loop of isolated lactic acid bacteria (unit: mm) Indication strain
Isolated lactic acid bacteria LAB1 LAB2 LAB3 LAB4 Candida albicans
( Candida albicans KCTC 17485) 10 - - - Aspergillus Flaubs
( Aspergillus flavus KCTC16682) 12 - 12 - Bacillus cereus
( Bacillus cereus KCCM12142) 22 20 20 24 Staphylococcus aureus
( Staphylococcus auerus KCCM 11335) 20 20 15 25 Escherichia coli ( E. coli KCCM11234) 19 15 15 20

<1-3> Identification of selected lactic acid bacteria

a. Morphological characteristics

The morphological characteristics of LAB1 were observed using a microscope. Morphological and biochemical characteristics were investigated according to Bergy's manual of systematic bacteriology and Gram staining was performed.

As a result, it was confirmed that the isolated strain was a gram-positive strain and had a bacterium form. The colonies were light yellow opaque and the surface of the colonies was smooth (Table 2).

characteristic Lactobacillus platarium DSR KF14 Gram + Colony form Rod Colony color yellow Motility - Catalase (Catalase) - Fermentation form homo Spore - Growth at 16 ℃ + Growth in 4% NaCl +

b. Biochemical property

LAB1 was identified by an API system (La Balme-les-Grottes, France). First, the sterilized platinum-fungus colonies were each taken and suspended in 2 ml of sterile distilled water. The bacterial solution was suspended in 5 ml of sterilized distilled water at a concentration of MccFaland Standard Solution No. 2 supplied by API 50CH kit (BioMerieux, France). The supernatant was added to the liquid medium of the API 50CH kit and homogenized, and then 200 μl of each of the 50 tubes of the API 50CH kit was inoculated. The tube was covered with mineral oil and then cultured at 30 DEG C for 48 hours. The culture solution was analyzed by an API system to identify 49 types of carbohydrate fermentation patterns, and the results were entered into an ATB identification computer system.

As a result, the LAB1 lactic acid bacteria selected in Example <1-2> were found to belong to the Lactobacillus genus having the following carbohydrate fermentation pattern (Table 3).

Control group - D-Manno + Salincin + Gentio Vioz + Glycerol - L-Solbos - Cellobiz + D-Turanz - Earl's tall - Rannoz - Malts + D-rings - D-arabinose - Dissytol - Laktos + D-Tagatoz - L-arabinose + Inositol - Melibiose + D-fucose - Riboze + Mannitol + Saka Rose + L-fucose - D-xylose + Sorbitol - Trehalose - D-arabitol - L-xylose - D-mannoside - Inulin - L-arabitol - Adonitol - D-glucoside + Melrietto + Gluconate + Xyloside - Glucosamine + D-Rapinoz - 2-Setto-gluconate - Galactos + Amigalline + Amidon - 5-Seto-gluconate + D-glucose + Arbutin + Glycogen - D-Prostose + Esculin + Xylitol +

c. 16S rDNA sequencing

The base sequence of 16S rDNA of the selected LAB1 lactic acid bacteria was analyzed according to a conventional method known in the art. As a result, it was found that the 16S rDNA sequence (SEQ ID NO: 1) of the LAB1 lactic acid bacteria was 99% identical to the 16S rDNA sequence of Lactobacillus sacae. Therefore, the present inventors designated the LAB1 lactic acid bacteria as "Lactobacillus casei DSR KF14 " and deposited it on the Korean Culture Center of Microorganisms on Oct. 30, 2014 (Accession No .: KCCM11595P).

&Lt; 1-4 >

a. Test for antibacterial activity

To test the antimicrobial activity of the Lactobacillus casei DSR KF14 strain, a third subculture was carried out on a MRS liquid medium. Nutrient agar was supplemented with pathogenic bacterium Bacillus cereus KCCM 12142, Staplyococcus aureus aureus KCCM11335, Listeria monocytogenes KCCM40307, E. coli KCCM11234, Salmonella typimurium KCTC2057, and Streptococcus mutans KCTC3065 were each inoculated at 1% It was hardened on the plate and then dried for 20 minutes. The culture supernatant of Lactobacillus casei DSR KF14, which had a diameter of 9 mm, was poured through a hole with a diameter of 9 mm and then incubated at 37 ° C for 24 hours to measure the size of the inhibitory loop (Table 4).

b. Antifungal test

For the anti-yeast test of the Lactobacillus casei DSR KF14, Candida albicans KCTC 17485 and Zygosaccharomyces rouxii yeast were cultured in a third subculture in a YPD liquid medium, and 1% was inoculated in a YPD agar medium and hardened in a plate And dried for 20 minutes. 50 占 퐇 of a culture supernatant of Lactobacillus casei DSR KF14, which had been punched through a hole having a diameter of 9 mm, was poured into a cockbearer, and the mixture was incubated at 25 占 폚 for 48 hours to measure the size of the inhibitory circle. .

To test for the antifungal activity of the Lactobacillus casei DSR KF14, spores were prepared by inoculating Aspergillus flavus KCTC16682, Penicillium expensum KCTC6434, and Fusarium oxysproum KCTC16909 fungi into the PDA medium by the above method. The extracted spores were inoculated 1%, hardened on the plate, and dried for 20 minutes. 100 μl of a culture supernatant of Lactobacillus casei DSR KF14, which had been punched through a hole having a diameter of 9 mm, was poured into a cockbearer, and the suspension was cultured at 25 ° C for at least 48 hours. The size of the inhibitory loop was measured and shown in Table 4 .

As shown in Table 4 below, the selected lactic acid bacteria Lactobacillus casei DSR KF14 exhibited antibacterial activity against yeast and fungi in comparison with the same kind of lactic acid bacteria, and thus it was confirmed that the antibacterial spectrum was broad.

Antifungal and antibacterial test results (Unit: mm) Indication strain Lactobacillus SA
DSR KF14 Lactobacillus casei KCCM40264 Lactobacillus casei KCCM40275 Candida albicans KCTC17485 13 - - Zygosaccharomyces rouxii 13 - - Aspergillus flavus KCTC16682 12 - 13 Penicillium expensum KCTC6434 13 - - Fusarium oxysproum KCTC16909 14.5 10 - Bacillus cereus KCCM12142 38 30 30 Staphylococcus auerus KCCM11335 37 39 29 Listeria monocytogenes KCCM40307 38 39 38 E. coli KCCM11234 41 35 31 Salmonella typhimurium KCTC2057 30 22 25 Streptococcus mutans KCTC3065 24 18 20

<1-5> Acid resistance  Test

Lactobacillus saccharum DSR KF14, Lactobacillus saccharum KCCM40264 and Lactobacillus saccharum KCCM40275 were added to 5 ml of an MRS liquid medium adjusted to pH 2.5, 3.0, 3.5, 4.0, 4.5, 5.0 using lactic acid Respectively. The cells were incubated at 25 DEG C for 72 hours to observe whether the cells were growing or not.

As a result, as shown in Table 3, it was confirmed that the selected lactic acid bacteria Lactobacillus casei DSR KF14 had a survival rate of 90% at a pH of 2.5 and was active for growth.

Results of acid resistance test (unit: survival rate (%)) pH 2.5 3.0 3.5 4.0 4.5 5.0 Lactobacillus case DSR KF14 90 95 99 100 100 100 Lactobacillus casei KCCM40264 20 50 88 90 100 100 Lactobacillus casei KCCM40275 55 75 90 98 100 100

<1-6> My bile  Test

Lactobacillus casei DSR KF14, Lactobacillus casei KCCM40264 and Lactobacillus casei KCCM40275 were inoculated in 10 ml of MRS liquid medium adjusted to 0.3%, 0.5%, 1% and 3% of oxgall, respectively. Then, the growth of the strain at the artificial bile concentration was observed while culturing at 37 캜 for 2 hours.

As shown in the following Table 6, the selected lactic acid bacteria Lactobacillus casei DSR KF14 was active in oxgall 0.3% and 0.5% and exhibited a high survival rate of 100% even in 3% Strong.

Results of my biliary test (unit: survival rate (%)) Oxgall concentration (%) 0.3 0.5 One 3 Lactobacillus case DSR KF14 100 100 100 100 Lactobacillus casei KCCM40264 95 85 50 40 Lactobacillus casei KCCM40275 98 95 80 70

< Manufacturing example  1> Production of Kimchi Using Lactic Acid Bacteria of the Present Invention

The cabbage was harvested with salt so that the cabbage had a salinity of 2.3%. (2.5 wt.%), Garlic (2 wt.%), Ginger (0.5 wt.%), Green onion (2 wt.%), Rape seeds (9 wt.%) And sugar Kimchi seasoning was mixed. Thereafter, the plant lactobacillus lactobacillus sake of the present invention as a starter was added in an amount of 1 wt% (about 15 x 10 strains) based on the total weight of the kimchi. Lactobacillus saccharum cells were cultured in MRS medium at 37 ° C for 24 hours, centrifuged at 10,000 g for 5 minutes (4 ° C), and the pellet was washed twice with sterilized water. Thereafter, the prepared kimchi was aged at 10 DEG C for 6 days.

< Manufacturing example  2> Production of feed using lactic acid bacteria of the present invention

2.0% of the Lactobacillus saccharum DSR KF14 lactic acid bacteria of the present invention was inoculated into a medium containing 2.0% glucose, 1.0% peptone, 1.0% yeast extract, 0.2% phytic acid, 0.05% magnesium sulfate, 0.05% cysteine, And aerobically cultured at 37 ° C for 2 days. The culture medium of the strain and the excipient degreasing solution were simply mixed at a weight ratio of 1: 1 and then dried at 40 ° C or lower. The dried mixture was pulverized to prepare a feed composition. At this time, the prepared feed composition was prepared so that the lactic acid bacterium of the present invention contained 1 x 10 &lt; 6 &gt; cfu / g or more and mixed with a conventional feed additive.

< Manufacturing example  3> Production of Pharmaceuticals Using Lactic Acid Bacteria of the Present Invention

2.0% of the lactic acid bacteria Lactobacillus saccharum DSR KF14 of the present invention was inoculated into a vegetable lactic acid bacteria medium containing 20% by weight of the plant, 10% by weight of Chinese cabbage, 3% by weight of glucose, 2% After rapid fermentation at -70 ° C or lower, the pharmaceutical composition was prepared by freeze-drying at -60 ° C for 36 hours. At this time, the temperature of the hot plate for increasing the efficiency of moisture evaporation during lyophilization was set to 30 ° C, and the pharmaceutical composition prepared at this time included lactic acid bacteria of the present invention at 1 × 10 6 cfu / g or more.

Each of the above-prepared pharmaceutical compositions was molded into tablets, pills, and capsules according to a conventional method to prepare pharmaceutical preparations.

< Manufacturing example  4> Production of Cosmetic Products Using Lactic Acid Bacteria of the Present Invention

The ingredients were weighed in order according to the compounding ratio below, uniformly mixed, and the remaining amount was filled with purified water to prepare a softening longevity (skin) containing the lactic acid bacterium of the present invention.

Flexible longevity weight (%)

Purified water balance

Glycerin 21,3

Butylene glycol 2

Citric acid 0.01

Ethanol 10

Preservative 0.0001

Pigment 0.0001

Fragrance 0.0001

Lactic acid fermentation broth 10

The lactic acid bacteria Lactobacillus saccharum DSR KF14 according to the present invention has antifungal activity as well as antifungal activity, and thus can be effectively used for sterilization, antibacterial and preservation purposes in various fields such as foods, feeds, medicines and cosmetics.

Korea Microorganism Conservation Center (overseas) KCCM11595P 20141030

<110> DAESANG FNF CORPORATION <120> LACTIC ACID BACTERIUM SEPARATED FROM KIMCHII AND HAVING          ANTIFUNGAL ACTIVITY AND COMPOSITON INCLUDING IT <130> 14-9903 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1531 <212> DNA <213> Lactobacillus sakei <400> 1 cagatcgac gcactctcgt tagattgaag aagcttgctt ctgattgata acatttgagt gagtggcgga 120 cgggtgagta acacgtgggt aacctgccct aaagtggggg ataacatttg gaaacagatg 180 ctaataccgc ataaaaccta gcaccgcatg gtgcaaggtt gaaagatggt ttcggctatc 240 actttaggat ggacccgcgg tgcattagtt agttggtgag gtaaaggctc accaagaccg 300 tgatgcatag ccgacctgag agggtaatcg gccacactgg gactgagaca cggcccagac 360 tcctacggga ggcagcagta gggaatcttc cacaatggac gaaagtctga tggagcaacg 420 ccgcgtgagt gaagaaggtt ttcggatcgt aaaactctgt tgttggagaa gaacgtattt 480 gatagtaact gatcaggtag tgacggtatc caaccagaaa gccacggcta actacgtgcc 540 agcagccgcg gtaatacgta ggtggcaagc gttgtccgga tttattgggc gtaaagcgag 600 cgcaggcggt ttcttaagtc tgatgtgaaa gccttcggct caaccgaaga agtgcatcgg 660 aaactgggaa acttgagtgc agaagaggac agtggaactc catgtgtagc ggtgaaatgc 720 gtagatatat ggaagaacac cagtggcgaa ggcggctgtc tggtctgtaa ctgacgctga 780 ggctcgaaag catgggtagc aaacaggatt agataccctg gtagtccatg ccgtaaacga 840 tgagtgctag gtgttggagg gtttccgccc ttcagtgccg cagctaacgc attaagcact 900 ccgcctgggg agtacgaccg caaggttgaa actcaaagga attgacgggg gcccgcacaa 960 gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacatc 1020 ctttgaccac tctagagata gagctttccc ttcggggaca aagtgacagg tggtgcatgg 1080 ttgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttat 1140 tactagttgc cagcatttag ttgggcactc tagtgagact gccggtgaca aaccggagga 1200 aggtggggac gacgtcaaat catcatgccc cttatgacct gggctacaca cgtgctacaa 1260 tggatggtac aacgagtcgc gagaccgcga ggtttagcta atctcttaaa accattctca 1320 gttcggattg taggctgcaa ctcgcctaca tgaagccgga atcgctagta atcgcggatc 1380 agcatgccgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accatgagag 1440 tttgtaacac ccaaagccgg tgaggtaacc ttcgggagcc agccgtctaa ggtgggacag 1500 atgattaggg tgaagtcgta acaaggtaac c 1531

Claims (7)

Lactobacillus sakei strain DSR KF14 (accession number: KCCM11595P) having antifungal and antibacterial activity. [Claim 2] The method according to claim 1, wherein the strain has acid resistance and biliary cholesterol. Lactobacillus &lt; RTI ID = 0.0 &gt; sakei) DSR KF14 strain. The method of claim 1, wherein the fungi are selected from the group consisting of Candida, Zygosaccharomyces, Aspergillus, Penicillium, and Fusarium. &Lt; RTI ID = 0.0 &gt; Lactobacillus &lt; / RTI &gt; sakei) DSR KF14 strain. An antifungal and antibacterial composition comprising the strain of any one of claims 1 to 3 or a culture thereof. A food comprising the antifungal and antimicrobial composition of claim 4. A cosmetic comprising the antifungal and antimicrobial composition of claim 4. A method for producing a fermented food product using the strain of any one of claims 1 to 3 or a culture thereof as a starter.