KR101970148B1 - Novel Lactobacillus sakei KR18 with probiotic activities - Google Patents

Abstract

The present invention relates to a lactobacillus sakei KR18 (KCCM 12350P) with probiotic activity by having excellent milk protein resolution and antibacterial activities; and a composition containing the same. The lactobacillus sakei KR18 (KCCM 12350P) of the present invention is able to: improve probiotic functionality by increasing the survival of lactic acid bacillus in intestines with excellent acid resistance and bile resistance; and be used as a starter to produce fermented milk using milk or soymilk through the curd formation of milk protein and soymilk protein. Moreover, the lactobacillus sakei KR18 (KCCM 12350P) of the present invention has excellent antibacterial properties against pathogenic bacteria and thus, improves intestinal flora by suppressing the proliferation of harmful bacteria when used as a probiotic substance. Moreover, the lactobacillus sakei KR18 (KCCM 12350P) of the present invention has excellent milk protein resolution and can be used as lactic acid bacillus probiotic substance having excellent intestinal attachment ability. The lactobacillus sakei includes a sequence listing represented in sequence number 1 as 16S rRNA and shows a positive reaction to D-galactose and lactose.

Description

Novel Lactobacillus saccharum strain KR18 having a probiotic activity and a composition containing the same - " Novel Lactobacillus sakei KR18 with probiotic activities &

The present invention relates to novel Lactobacillus four K (Lactobacillus sakei) relates to KR18 (KCCM 12350P), and more particularly Salmonella (Salmonella) genus, Staphylococcus aureus (Staphylococcus aureus), and Escherichia coli (E-coli) and so on for the pathogenic bacteria Lactobacillus sakei KR18 (KCCM 12350P) having antibacterial activity and probiotic activity excellent in milk protein degradation ability and compositions containing the same.

Probiotics is a living organism that enters the body and gives a healthful effect. It plays an important role in maintaining the balance of normal intestinal flora and inhibiting the growth of pathogenic bacteria. Most probiotics known to date are lactic acid bacteria. In order for bacteria such as lactic acid bacteria to be recognized as probiotics, they must survive in stomach and bile acid to reach the small intestine and proliferate and settle in the intestines. They should show useful effects in the intestine, There is a need to be.

Probiotics have been shown to improve the intolerance of lactose, lower cholesterol and blood pressure, improve immune function, prevent infection and so on. Probiotic products have been consumed as fermented milk products made from lactic acid bacteria, and recently they have also been developed in the form of granules and powders. Accordingly, it is necessary to develop probiotics having excellent functionality and good usability.

Korean Patent No. 10-1541280 (Registration Date 2015.07.28) discloses that Lactobacillus sp., Which inhibits the epithelial cell adhesion of Escherichia coli 0157: H7 bacterium, which simultaneously produces shiga toxin type 1 and type 2, ) KU4 (KCCM 10975P) and lactic acid bacteria probiotics containing the same.

The present invention relates to an antimicrobial activity against pathogenic bacteria and an excellent And to develop and provide a food composition and a pharmaceutical composition having probiotic activity.

The present invention relates to a Lactobacillus sakei containing 16S rRNA and having a nucleotide sequence as set forth in SEQ ID NO: 1, wherein the lactic acid bacterium is characterized by exhibiting a positive reaction to D-galactose and lactose, to provide.

In the lactic acid bacteria of the present invention, the Lactobacillus sakei may preferably be Lactobacillus sakei KR18 (KCCM 12350P).

In the lactic acid bacteria of the present invention, the Lactobacillus sakei may preferably be one having at least one property selected from among acid resistance, bile resistance, antibacterial activity, milk protein degradation ability and intestinal adhesiveness.

Meanwhile, the present invention provides a food composition characterized by containing Lactobacillus sakei KR18 (KCCM 12350P) as an active ingredient.

In the food composition of the present invention, the food composition may be any one selected from fermented milk of lactic acid bacteria, soybean milk, powdered milk, beverage or health supplement.

Lactobacillus sakei KR18 (KCCM 12350P) selected in the present invention is excellent in acid resistance and biliary excretion, thereby enhancing the intestinal viability of the lactic acid bacteria, thereby improving the probiotic function and inhibiting the curdling of milk protein and soy protein Can be used as a starter for the production of fermented milk using milk and soy milk.

In addition, Lactobacillus sakei KR18 (KCCM 12350P) of the present invention exhibits excellent antibacterial properties against pathogenic bacteria, and when used as a probiotic agent, inhibits the proliferation of harmful bacteria and can improve the intestinal flora.

Further, Lactobacillus sakei KR18 (KCCM 12350P) of the present invention is excellent as a milk protein degrading ability and can be utilized as a lactic acid bacteria probiotics having excellent adherence ability.

1 is a microscopic photograph of Lactobacillus sakei KR18 (KCCM 12350P) of the present invention and a photograph of Lactobacillus sakei KR18 (KCCM 12350P) and a comparative strain CSLA ( Lactobacillus acidophilus CSLA) It is a photograph of the experiment result of whether or not the transparent ring showing milk protein decomposition ability is generated.
FIG. 2 is a diagram showing a phylogenetic tree of Lactobacillus sakei KR18 (KCCM 12350P) of the present invention.
FIG. 3 is a photograph showing the optimum growth temperature of Lactobacillus sakei KR18 (KCCM 12350P) of the present invention cultured at 35 ° C and 45 ° C in BCP agar.
FIG. 4 is a graph and a graph showing the growth rate, growth curve, and pH change of Lactobacillus sakei KR18 ( Lactobacillus sakei KR18 (KCCM 12350P)) and the comparative strain CSLA of the present invention.
FIG. 5 is a table and photographs showing experimental results showing the sugar disintegration characteristics of the Lactobacillus sacrifia KR18 ( Lactobacillus sakei KR18 (KCCM 12350P)) using the API kit of the present invention.
FIG. 6 is a photograph of experimental results on the curd formation characteristics of the milk protein and soybean protein of Lactobacillus sakei KR18 ( Lactobacillus sakei KR18 (KCCM 12350P)) of the present invention and the comparative strain CSLA.
FIG. 7 is a photograph of the results of an antibacterial activity test for five pathogenic strains of Lactobacillus sakei KR18 ( Lactobacillus sakei KR18 (KCCM 12350P)) and a comparative strain CSLA of the present invention.
FIG. 8 is a table and a graph showing the results of measurement of milk protein degradation activity of Lactobacillus sacrifia KR18 ( Lactobacillus sakei KR18 (KCCM 12350P)) of the present invention. Milk protein degradation was expressed as free tyrosine content.
9 is an experimental result table and a graph showing the intestinal adherence ability of Lactobacillus sakei KR18 (KCCM 12350P) and the comparative strain L. rhamnosus GG (LGG).

The present invention relates to a Lactobacillus sakei containing 16S rRNA and having a nucleotide sequence as set forth in SEQ ID NO: 1, wherein the lactic acid bacterium is characterized by exhibiting a positive reaction to D-galactose and lactose, to provide.

In the present invention, excellent lactic acid bacteria were screened from the fermented kimchi prepared by the conventional method, and it was judged that the milk protein decomposition ability to produce a transparent ring through a paper disc assay for selected colonies was investigated. Physiological, biochemical and antimicrobial activity tests were conducted to select and isolate the most excellent strains, thereby completing the present invention.

Lactic acid bacteria are bacteria that decompose saccharides such as glucose to produce lactic acid. They are also referred to as lactic acid bacteria, and they ferment sugars to obtain energy. Lactobacillus and Streptococcus sp. Are known to be typical lactobacillus, and they are used in the manufacture of dairy products (yogurt, cheese, etc.), kimchi, brewed foods (such as sake, soybean paste, and soy sauce).

In the lactic acid bacteria of the present invention, the Lactobacillus sakei may be preferably Lactobacillus sakei KR18 (KCCM 12350P).

In the lactic acid bacteria of the present invention, Lactobacillus sakei may preferably be one having at least one property selected from among acid resistance, bile resistance, antibacterial activity, milk protein degradation ability and intestinal adhesiveness.

The lactic acid bacteria of the present invention can be used as a starter for the production of fermented milk using milk and soybean milk by observing the curd formation of milk protein and soybean protein when the following experiment is conducted and it was confirmed that Salmonella sp. Philo nose in Syracuse (Staphylococcus sp.), Bacillus (Bacillus sp.), Escherichia coli and exhibits a superior antimicrobial properties against a pathogen, such as (Escherichia coli) inhibited the growth of a probiotic drive harmful bacteria the effect of improving the intestinal microflora It can be demonstrated. In addition, the cleavage ability of milk protein was confirmed through the production of a transparent ring in the skim milk-containing medium, and it was confirmed that the milk protein degradation ability was excellent by measuring the amount of tyrosine released during the fermentation process, and thus the lactic acid bacteria probiotics It can be used.

Meanwhile, the present invention provides a food composition characterized by containing Lactobacillus sakei KR18 (KCCM 12350P) as an active ingredient.

In the food composition of the present invention, for example, it may be any one selected from fermented milk of lactic acid bacteria, soybean milk, milk powder, beverage or health supplement, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[Example 1: Selection and isolation of Lactobacillus saccharum KR18 excellent in milk protein degradation ability]

In this Example, a strain having excellent milk protein degradation ability was selected and identified from traditional fermented kimchi.

1) Selection of Lactobacillus saccharum KR18 with excellent milk protein degradation and morphological and biochemical characterization

Lactic acid bacteria were isolated from traditional fermented kimchi in Gangwon province. 10 g of each sample was diluted in sterilized water and plated on a MRS (Difco Laboratories, Detroit, MI, USA) solid medium. For a period of time, and 50 colonies were firstly selected for the growth of milky white colonies. The selected colonies were again inoculated into BCP agar, and LCPs were identified as lactic acid bacterium which formed yellow by lactic acid production. Of these, 32 colonies having excellent acid production were separated pure. Then, a 6 mm paper disc was placed on the MRS plate medium mixed with 2% reduced skim milk, and then 10 μl of the separated lactic acid bacteria culture liquid was added dropwise thereto and cultured at 37 ° C for 48 hours. At this time, the isolate producing the transparent ring was temporarily determined to have milk protein degradation ability. For comparison, the commercial strain, CSLA ( Lactobacillus acidophilus CSLA), was used in the experiment. Compare strain CSLA (Lactobacillus acidophilus CSLA) and a clear zone results confirm whether to generate, starting with excellent strain KR18 strain thereby forming a clear zone of greater about 10mm than about 8mm of CSLA exhibits a superior milk proteolytic characteristics as shown in Figure 1 Physiological, biochemical and microscopic observations were performed.

As a result of analyzing the morphological and biochemical characteristics of the isolated strain (KR18), the results were as shown in Table 1 and FIG. 1, and it was confirmed that the isolated strain (KR18) had a rod- Gram stain revealed Gram - positive bacteria. Catalase test showed catalase negative reaction, and it was confirmed that it proliferated regardless of presence or absence of oxygen. Thus, it was found that the isolated strain (KR18) had typical lactic acid bacterial characteristics.

characteristic result Morphology rod Gram staining (+/-) + Catalase test (+/-) - Aerobic / Anaerobic + / +

2) Genetic identification of Lactobacillus saccharum KR18 with excellent milk protein degradation

Genomic identification was carried out by 16S rRNA gene analysis for accurate identification of the isolate (KR18).

For amplification of the 16S rRNA of a colony sample of the isolate strain KR18, a primer for lactic acid bacteria, forward primer 27F (5'-aga gtt tga tcc ctc ag-3 ') and reverse primer 1492R (5'-ggt tac ctt gtt acg act t-3 '), the first cycle was 15 minutes at 95 ° C, 30 cycles were at 20 seconds at 95 ° C, 40 seconds at 50 ° C, 1 minute and 30 seconds at 72 ° C, PCR was carried out for 10 min at RT for 10 min. The PCR products were amplified by electrophoresis, and genomic identification was carried out based on sequencing analysis using NCBI genebank. In case of strains showing 97% or more nucleotide sequence homology in general taxonomy, they can be defined as one species. The analysis results are as follows. Lactobacillus sakei was identified as 99% or more homology as shown in Table 2 below. The phylogenetic tree of the identified strain (KR18) can be shown in FIG. 2, and the 16S rRNA nucleotide sequence used is shown in Table 3 below.

Strain No. Identification 16S rRNA 27F 1492R KR18 Lactobacillus sakei 99% 100%

designation 16S rRNA sequence Identification strain (KR18) SEQ ID NO: 1

The inventors of the present invention deposited Lactobacillus sakei KR18 (KCCM 12350P) in the Korean Microorganism Conservation Center.

[Experimental Example 1: Experiment to confirm optimal growth temperature and growth characteristics of novel Lactobacillus saccharum KR18]

In this experiment, experiments were conducted to determine the optimal growth temperature and growth characteristics of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P).

In order to confirm the optimum growth temperature of the isolated strain (KR18), streaking on BCP agar and cultivation at 45 ° C and 35 ° C for 24 hours resulted in almost no growth at 45 ° C, It was found that it was mesophilic because it grows well.

On the other hand, to confirm the growth characteristics of the isolate (KR18), the degree of propagation of the strain was measured by the number of viable cells and pH. As a comparative strain, commercial strain CSLA ( L. acidophilus ) was used. Each strain was inoculated into MRS broth and incubated at 37 ° C for 12 hours. The samples were taken at 2-hour intervals and the growth curve was prepared by measuring the absorbance at 660 nm using a spectrophotometer (X-ma 1200, Human corporation) The change in pH was measured using a pH meter (Orion 3 Star, Thermo Scientific).

 As a result, as shown in FIG. 4, the growth of the strain showed a logarithmic growth period from 6 hours to 10 hours after the incubation, and it was confirmed that the strain was entering a period of stagnation after a gradual increase after 10 hours. The growth patterns of isolates (KR18) and control strains (CSLA) showed similar trends, indicating that the isolate (KR18) had excellent growth ability as the control strain CSLA. In addition, the change of pH showed a rapid decrease between 6 hours and 10 hours in proportion to the growth curve of the isolates and the control bacteria, and it was confirmed that the pH could be applied to the production of fermented milk at the pH of 4.5.

[Experimental Example 2: Test for confirming acid resistance and biliary properties of novel Lactobacillus saccharum KR18]

In this experiment, culture experiments were carried out to confirm the acid resistance and biliary cholesterol of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P).

In order to confirm acid resistance, the isolate strain (KR18) was inoculated on MRS broth and incubated at 37 ° C for 24 hours. The culture broth was inoculated with 2% each of pH 3.0 MRS broth prepared by 1N-HCl correction and incubated at 37 ° C for 3 hours . The cultured samples were decanted into sterilized water and 1 ml was taken, inoculated on MRS agar, and cultured at 37 ° C for 24 hours. The resulting colonies were counted, and the resistance to acidic pH was determined by comparing the number of colonies after initial incubation with that after incubation Respectively. As a result, the survival rate was 60.1% as compared with the initial number of bacteria as shown in Table 4, which was lower than that of L. acidophilus , which is generally known to have an acid resistance of 90% or more at pH 2.5-3.5. I confirmed it. Considering that the general fermented milk has a pH of 4.0 or more, it was judged that KR18 could be used as a fermented milk starter.

Next, the isolate (KR18) was inoculated into the MRS broth and cultured at 37 ° C for 24 hours. The broth was inoculated with 2% each of 0.3% oxgall bile salt-added MRS broth at 37 ° C Time culture method. The cultured samples were decanted into sterilized water and 1 ml was taken, inoculated on MRS agar, and cultured at 37 ° C for 24 hours. The resulting colonies were counted, and the resistance to acidic pH was determined by comparing the number of colonies after initial incubation with that after incubation Respectively. As a result, as shown in Table 4, the number of viable cells was 226.9% as compared with the initial number of bacteria when cultured for 6 hours in the bile extract 0.3% added group. It was confirmed that the growth of isolated strain (KR18) was not inhibited at a concentration of 0.3% of Bile extract. The isolate strain KR18 increased the viability of the lactic acid bacteria in the fermented milk and could enhance the probiotic function.

sample Acid resistance (pH3.0 / 3hr) My bile (bile salt 0.3% / 6hr) L. sakei KR18 60.1% 226.9%

[Experimental Example 3: Confirmation test for the sugar disintegration properties of novel Lactobacillus saccharum KR18]

In this experiment, 49 sugar availability pattern analysis and identification were carried out using the API kit to identify the lactic acid characterization of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P).

Each isolate (KR18) was inoculated on MRS agar and cultured at 37 ° C for 24 hours. The colonies were aseptically collected and transferred to API 50 CHL medium (BioMeriex Co., Lyon, France) The suspension was dispensed in an API 50 CH strip and cultured at 37 ° C for 48 hours. Strip readings were negative (-) and positive (+) due to the color change of each tube, and were identified using the API 50 CHL database. As a result, as shown in FIG. 5, it was confirmed that the isolated strain (KR18) showed a positive reaction in D-galactose and lactose.

[Experimental Example 4: Experiment to confirm curd formation characteristics of milk protein and soybean protein of new lactobacillus sacrifice KR18]

Experiments were conducted to determine the curd formation characteristics of the fermented milk protein and the soybean protein of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P).

The isolate strain KR18 was cultured in MRS broth at 37 ℃ for 24 hours and then sterilized at 85 ℃ for 10 minutes with 1% (w / w) skim milk and 8% (w / w) Respectively. As a control, CSLA ( L. acidophilus ), a commercial strain, was cultured in MRS broth for 12 hours and then inoculated 1% in a similar manner. The culture was incubated at 37 DEG C for 12 hours to observe the degree of formation of curd and determine the possibility of use as a starter for the production of fermented milk.

The results of incubation were as shown in Table 5 and FIG. 6, and soybean protein began to form curds from 5 hours after culturing and curd formation was observed after 8 hours from milk protein. The isolate strains (KR18) showed slightly slower acid production than the CSLA (commercial strains), but showed a similar level of curd formation as a whole at 12 hours of culture, indicating that they could be used as starters for the production of fermented milk using milk and soy milk Respectively.

division 10% skim milk Soybean oil (TS 8%) CSLA KR18 CSLA KR18 Curd formation O O O O pH 3.92 4.32 3.68 3.67 Acidity (%) 0.94 0.74 0.50 0.55

[Experimental Example 5: Test for confirming antimicrobial activity of novel Lactobacillus saccharum KR18]

In this experiment, an experiment was conducted to confirm the antibacterial activity of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P).

Antimicrobial activity test of the strain (KR18) are Paper disk agar by using a diffusion method S. typhimurium (Salmonella typhimurium), L. monocytogenes to Ness (Listeria monocytogenes), Staphylococcus aureus (Staphylococcus aureus), Bacillus cereus (Bacillus cereus), E. coli (Escherichia coli) for 24 hours in a 37 ℃ after inoculation each tryptic soy broth (Oxoid, UK) with pathogenic microorganisms, such as the culture was used as a test strain. The isolated lactic acid bacteria culture broth was inoculated into MRS broth and pre-cultured at 37 ° C for 12 hours and used as a sample. CSLA ( Lactobacillus acidophilus CSLA) were preincubated in the same manner and compared. Each test strain was inoculated on a tryptic soy agar plate and poured into a pre-made tryptic soy agar plate for 30 minutes. Then, the sterilized paper disk was placed on the sterilized soybean agar plate, and 50 μl of each lactic acid bacterial culture sample was added dropwise and cultured at 37 ° C. for 24 hours And the formation of a clear zone was confirmed. The formation of the inhibitory ring formed inhibitory rings for all five pathogenic bacteria. The positive inhibitory effect was greater than 10 mm, while the inhibitory effect was less than 10 mm.

As a result, as shown in FIG. 7, the isolate strain (KR18) exhibited antibacterial activity against all five pathogenic strains tested and exhibited superior antimicrobial properties to CSLA against four other pathogenic bacteria except listeria. Generally, lactic acid bacteria are known to inhibit the growth of pathogenic bacteria by lowering the acidity by producing lactic acid, and to produce antimicrobial substances such as bacteriocin. Therefore, the isolate strain (KR18), which has excellent antimicrobial activity, is expected to have the effect of suppressing the growth of harmful microorganisms and improving the intestinal microflora when used as a probiotic.

[Experimental Example 6: Experiment to confirm the milk protein decomposition (free tyrosine content) of novel Lactobacillus sacrifix KR18]

Experiments were conducted to confirm the milk proteolytic potency of the selected isolate strain Lactobacillus sakei KR18 (KCCM 12350P) by the amount of free tyrosine produced during fermentation.

The milk protein digestibility of isolate strain KR18 was determined by using 10% reduced skim milk as the medium and the free tyrosine content. Samples were taken at 4 - hour intervals while incubated at 37 ℃ for 24 hours after inoculating 1% of subcultured strains in sterilized reduced skim milk medium. The sample was filtered through whatman (No. 2) filter paper, and 5 mL of the filtrate was equilibrated with 0.44 M trichlroacetic acid (TCA). After allowing to stand for 30 minutes, filter with whatman (No. 42) filter paper, mix 1 mL of filtrate and 2.5 mL of 0.44 M sodium carbonate (Na ₂ CO ₃ ), add 200 μL of 1 N folin reagent and shake well for 5 minutes to remove blue It was developed. The absorbance at 540 nm was measured with a spectrophotometer (X-ma 1200), and the amount of tyrosine released was calculated as the value of tyrosine standard.

As shown in FIG. 8, when the isolated strain (KR18) was cultured at 37 ° C for 24 hours using a 10% reduced skim milk medium, the content of the released tyrosine was the highest in the range of 8 hours to 16 hours, It was confirmed that the protein showed excellent resolution of the milk protein as the paper disc assay.

[Experimental Example 7: Experiment for confirming adhesion of new Lactobacillus intestine cells]

In this experiment, experiments were conducted to confirm the ability of selected strains of Lactobacillus sakei KR18 (KCCM 12350P) to bind to the intestinal cells.

The adherence of intestinal cells is a probiotics and is considered to be a representative experiment to judge the suitability of the host for the intestinal environment. For the experiment, Caco-2 cell line, which is similar to the human small intestinal epithelium, was used and compared with commercial strains such as L. rhamnosus GG (LGG; Valio Ltd, Finland) · Since we have been screening, we also confirmed the relative adherence of LGG strain as a control in this experiment.

First, the Caco-2 cell line culture was incubated at 37 ° C in a 5% CO ₂ environment. The Caco-2 cell line between the 39th and the 41st cleavage site was used for adhesion measurements. caco-2 cells were loaded into 12 cell culture plate wells at a concentration of 5 × 10 ² ml and cultured for 72 hours to form a single cell layer filled with 80%. Caco-2cell was cultured in MEM medium without antibiotics for 2 hours and then mixed with 0.5 ml of the same volume of PBS solution containing lactic acid bacteria to be measured so that the final bacterial concentration was 2 × 10 ⁸ CFU / ml Respectively. The solution was then added to each tissue culture plate well and incubated for 2 hours at 37 ° C in 5% CO ₂ environment. After that, the single cell layer was washed twice with sterilized PBS (pH 7.2), fixed with 10% formalin solution, stained with crystal violet, and observed under a microscope. The viable cell count divided by initial cell count) x 100.

As a result, as shown in FIG. 9, when the LGG adherence of the LGG was 1.0, the isolated strain (KR18) showed a relatively low level of 0.7798.

As a result of the above results, the novel Lactobacillus sakei KR18 (KCCM 12350P) is excellent in acid resistance and biliary cholesterol, thereby enhancing the intestinal viability of the lactic acid bacterium, and exhibiting an effect of improving the probiotic function D-galactose and Lactose, and observed the curd formation of milk protein and soy protein. Thus, it could be used as a starter for the production of fermented milk using milk and soy milk.

 On the other hand, the strain of the present invention showed excellent antibacterial properties against pathogenic bacteria, and it was confirmed that the effect of improving the intestinal flora can be demonstrated by inhibiting the proliferation of harmful bacteria when used as a probiotic agent. In addition, it can be seen that a high content of free tyrosine can be utilized as a probiotic of lactic acid bacteria having excellent milk protein degradation ability and excellent adherence ability.

The inventors of the present invention deposited Lactobacillus sakei KR18 (KCCM 12350P) in the Korean Microorganism Conservation Center.

Name of depository: Korea Microorganism Conservation Center (overseas)

Accession number: KCCM12350P

Funding date: 20181029

<110> seoulfnb <120> Novel Lactobacillus sakei KR18 with probiotic activities <130> YP-18-238 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1433 <212> RNA <213> Lactobacillus sake <400> 1 actctcgttt agattgaagg agcttgctcc tgattgataa acatttgagt gagtggcgga 60 cgggtgagta acacgtgggt aacctgccct aaagtggggg ataacatttg gaaacagatg 120 ctaataccgc ataaaaccta acaccgcatg gtgtagggtt gaaagatggt ttcggctatc 180 actttaggat ggacccgcgg tgcattagtt agttggtgag gtaaaggctc accaagaccg 240 tgatgcatag ccgacctgag agggtaatcg gccacactgg gactgagaca cggcccagac 300 tcctacggga ggcagcagta gggaatcttc cacaatggac gaaagtctga tggagcaacg 360 ccgcgtgagt gaagaaggtt ttcggatcgt aaaactctgt tgttggagaa gaatgtatct 420 gatagtaact gatcaggtag tgacggtatc caaccagaaa gccacggcta actacgtgcc 480 agcagccgcg gtaatacgta ggtggcaagc gttgtccgga tttattgggc gtaaagcgag 540 cgcaggcggt ttcttaagtc tgatgtgaaa gccttcggct caaccgaaga agtgcatcgg 600 aaactgggaa acttgagtgc agaagaggac agtggaactc catgtgtagc ggtgaaatgc 660 gtagatatat ggaagaacac cagtggcgaa ggcggctgtc tggtctgtaa ctgacgctga 720 ggctcgaaag catgggtagc aaacaggatt agataccctg gtagtccatg ccgtaaacga 780 tgagtgctag gtgttggagg gtttccgccc ttcagtgccg cagctaacgc attaagcact 840 ccgcctgggg agtacgaccg caaggttgaa actcaaagga attgacgggg gcccgcacaa 900 gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacatc 960 ctttgaccac tctagagata gagctttccc ttcggggaca aagtgacagg tggtgcatgg 1020 ttgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttat 1080 tactagttgc cagcatttag ttgggcactc tagtgagact gccggtgaca aaccggagga 1140 cgtgctacaa 1200 tggatggtac aacgagttgc gagaccgcga ggtttagcta atctcttaaa accattctca 1260 gttcggattg taggctgcaa ctcgcctaca tgaagccgga atcgctagta atcgcggatc 1320 agcatgccgc ggtgaatacg ttcccgggcc tgtacacacc gcccgtcaca ccatgagagt 1380 ttgtaacacc caaagccggt gaggtaaccc ttcggggagc cagccgtcta agg 1433

Claims (5)

A Lactobacillus sakei containing 16S rRNA as a nucleotide sequence as set forth in SEQ ID NO: 1, wherein the Lactobacillus sakei is selected from the group consisting of Lactobacillus casei (D-galactose) and Lactose Lactobacillus sakei ) KR18 (KCCM 12350P).
delete The method according to claim 1,
The Lactobacillus sakei can be obtained by, for example,
Lactobacillus sakei KR18 (KCCM 12350P) having at least one property selected from acid resistance, biliary properties, antibacterial activity, milk protein degradation ability and intestinal adhesiveness.
A food composition characterized by containing Lactobacillus sakei KR18 (KCCM 12350P) as an active ingredient.
5. The method of claim 4,
The food composition may contain,
Lactic acid bacteria, fermented milk of lactic acid bacteria, soybean milk, powdered milk, or health supplements.

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