KR101933711B1 - Kimchi containing fermentation composition using novel lactic acid bacteria Lactobacillus plantarum DGK-17(KACC 81028BP) isolated from Kimchi and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a kimchi containing a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi and a preparation method thereof, and more particularly to a kimchi containing lactobacillus planta DGK-17 KACC 81028BP) was used to prevent the deterioration of the quality of kimchi caused by the administration of lactic acid-producing bacteria as well as the antimicrobial activity against the colonic cancer and antimicrobial activity against food poisoning bacteria at low concentrations. .

Description

Kimchi containing a fermented product prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi and a method of producing the kimchi containing the fermentation product of Lactobacillus plantarum DGK-17 (KACC 81028BP) from Kimchi and manufacturing method thereof}

The present invention relates to a kimchi containing a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi and a preparation method thereof, and more particularly to a kimchi containing lactobacillus planta DGK-17 KACC 81028BP) was used to prevent the deterioration of the quality of kimchi caused by the administration of lactic acid-producing bacteria as well as the antimicrobial activity against the colonic cancer and antimicrobial activity against food poisoning bacteria at low concentrations. .

Korea has the highest incidence of colorectal cancer in the world. Analysis of data from the World Health Organization (WHO) International Cancer Research Institute shows that the incidence of colorectal cancer in Korea is the highest among 184 countries surveyed, more than double the world average.

Increased colon cancer in modern humans includes high-calorie meats-oriented westernized eating habits, irregular eating habits, irritating food intake, and lack of exercise.

In particular, the eating habits have the greatest effect on the intestinal environment, that is, the balance of the intestinal bacteria. The immune system of the human body protects the beneficial bacteria and prevents the growth of the harmful bacteria so that the intestinal bacteria are balanced. When the immune system collapses, the harmful bacteria are overproduced and the toxins accumulate, causing digestive diseases and the like.

Colorectal cancer has no specific early symptoms, and most of the cases are already difficult to treat because of the large amount of cancer that has already developed.

Methods for treating colorectal cancer include chemical therapy for administering drugs such as anticancer drugs and radiation therapy for killing cells using radiation. However, since these treatments also act on the death of normal cells, hair loss, pain, anorexia, And serious side effects such as systemic fatigue are occurring.

In addition, due to the destruction of the immune function caused by erythrocyte destruction and erythrocyte destruction, serious diseases such as herpes zoster and herpes are incidentally generated, so that they can not escape the pain of cancer treatment other than the life extension effect.

The only way to solve this problem is to find ways to prevent and treat cancer by using food-borne materials.

In this way, there is a growing interest in health-promoting foods derived from plants and microorganisms that have immunity and anticancer efficacy that can be prevented and cured.

Lactic acid bacteria are defined as fermented crude oil or milk products with lactic acid bacteria. Fermented milk is prepared by adding lactic acid starter to crude oil, which is a complete food, and fermenting at a constant temperature. Lactobacillus acidophilus, Bifidobacteria, Streptococcus thermophilus, most of which are probiotics.

Probiotics is a microorganism that is safe as a GRAS (Generally Recognized As Safe), has acid resistance and biliary properties, and survives in the gastrointestinal tract. It is reported to settle in the intestines of the host and give the host a healthful effect.

The effects of probiotics include antibiotic-related diarrhea, intestinal infection by pathogenic bacteria, improvement of gastrointestinal tract diseases such as irritable bowel syndrome, reduction of atopic dermatitis, improvement of hypertension, decrease of blood cholesterol level and improvement of blood lipid status, anti-obesity effect, Studies on various functionalities such as anticancer effect on the liver have been carried out.

Kimchi which is a traditional fermented food in Korea is one of the most easily accessible foods. It has various and abundant lactic acid bacteria in fermentation process of kimchi, It is a report. Various health functionalities have been reported through research on the efficacy of kimchi, and various foods using kimchi lactic acid bacteria have been developed.

Korean Patent Registration No. 10-0526743 discloses a health supplement containing Kimchi as a main ingredient and a preparation method thereof, which is prepared by inoculation and fermentation of kimchi lactic acid bacteria in powdered cabbage and powdered into capsules and tablets.

Korean Patent No. 10-0977849 discloses a green juice composition containing Kimchi lactic acid bacteria and Korean Patent Publication No. 10-2016-0120875 discloses a fermented fruit drink using Kimchi lactic acid bacteria And a fermented fruit drink produced thereby.

Anticancer activity using lactic acid bacteria and probiotics has been reported so far, but its effect is very small.

In the case of kimchi, a method of additionally adding a probiotic agent to increase the number of lactic acid bacteria or to impart specific functionality has been proposed, but there is a limit to adversely affect the taste and quality of kimchi by the secondary fermentation of the probiotic agent.

Since the onset and prevention of cancer is very closely related to the diet, research that promotes health functionalities without deteriorating the taste and quality of everyday foods such as kimchi is a very necessary study at present.

Korea Patent No. 10-0526743 Korean Patent No. 10-0977849 Korean Patent Publication No. 10-2016-0120875

It is an object of the present invention to overcome the above-mentioned problems, and it is an object of the present invention to provide a fermented composition obtained by culturing a fermented composition obtained by culturing Lactobacillus plantarum DGK-17 (KACC 81028BP) And a method for manufacturing the same.

Another object of the present invention is to provide a kimchi excellent in anticancer activity against colon cancer and antibacterial activity against food poisoning bacteria even at a low concentration and a method for producing the same.

The fermentation composition prepared from the novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) of the present invention for solving the above-mentioned problems is prepared by mixing a seed culture of Lactobacillus plastarium DGK-17 (KACC 81028BP) At a concentration of 0.5 to 2% (v / v) in a medium containing 5 to 15 (w / v)% of at least one selected from the group consisting of wheat flour and fruit powder at 150 to 300 rpm, Cultivating the culture medium for a time, preparing the present culture medium, and concentrating the present culture medium.

The fermentation composition is centrifuged at 8,000 to 12,000 rpm for 5 minutes to 15 minutes at 4 ° C to recover the supernatant. The pH of the supernatant is adjusted to 6.5 to 7.5 and then lyophilized. And then concentrated under reduced pressure.

The kimchi containing the fermented composition is characterized by comprising 0.5 to 10% by weight of the fermented composition among 100% by weight of the whole kimchi.

The kimchi containing the above fermented composition is characterized by having anticancer activity.

The kimchi containing the fermented composition is characterized in that it has anticancer activity against the HCT116 colon cancer cell line.

The kimchi containing the fermented composition is characterized by having an antimicrobial activity against food poisoning bacteria.

The kimchi containing the fermented composition is characterized by having an antimicrobial activity against at least one of Pseudomonas aeruginosa 03K711 and Klebsiella pneumoniae 05K279.

A method for producing a kimchi comprising a fermented composition prepared from a Kimchi-derived lactic acid bacteria Lactobacillus plantarum DGK-17 (KACC 81028BP) of the present invention for solving the above-mentioned problems is characterized in that lactobacillus plantarum DGK-17 (KACC 81028BP (S100) for producing a fermented composition by culturing the fermented composition in a fermented state, and a step (S200) for preparing a kimchi by immersing the fermented composition in the fermented composition.

The fermentation composition preparation step (S100) comprises a seed culture step (S110) of producing a seed culture by laminating the Lactobacillus plantarum DGK-17 (KACC 81028BP) in an MRS medium and

The seed culture medium is inoculated at 0.5 to 2% (v / v) to a medium containing 5 to 15 (w / v)% of at least one of cereal powder, pulp powder and fruit powder, (S120) for culturing the present culture broth cultured at 40 ° C for 12 to 24 hours, and concentrating the culture broth (S130).

The concentration step (S130) comprises a centrifugation step (S131) of centrifuging the present culture broth at 8,000 to 12,000 rpm at 4 DEG C for 5 minutes to 15 minutes to recover the supernatant, and a pH adjusting step of adjusting the pH of the supernatant to 6.5 to 7.5 (S132); and a formulation step (S133) of preparing a pH-adjusted supernatant.

The formulation step (S133) may include a freeze-drying step (S133-1) of lyophilizing the pH-adjusted supernatant by pulverization or a low-temperature decompression step (S133-2) of concentrating the pH-adjusted supernatant under reduced pressure at 10 to 50 ° C And performs either one of the above.

The kimchi manufacturing step S200 includes preparing a pickled Chinese cabbage (S210), a seasoning preparation step (S220) of mixing the fermented composition and any seasonings, and a step S230 of filling the spices prepared in the pickled Chinese cabbage. And a fermentation step (S240) of fermenting the kimchi mixed with the sauce.

The seasoning preparation step (S220) is characterized by mixing 0.5 to 10% by weight of the fermented composition among 100% by weight of the whole kimchi.

As described above, according to the present invention, there is provided a kimchi comprising a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) and a preparation method thereof, (KACC 81028BP) is cultivated by using a fermented composition to prevent the deterioration of the quality of kimchi caused by the lactic acid-producing bacteria.

Further, according to the present invention, there is provided a kimchi containing a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi according to the present invention and a method for producing the same, It has an excellent antimicrobial activity.

1 is a graph depicting the inhibitory effect of selected fermentation compositions on selected target microorganisms according to an embodiment of the present invention.
FIG. 2 is a SEM photograph showing changes in cell morphology when treated with P. aeruginosa 03K711 and K. pneumoniae 05K279 of the fermentation composition.
FIG. 3 is a graph comparing the microbial community-forming ability of DGK-17. FIG.
FIG. 4 shows the MTT assay of HCT 1116 cell, a colon cancer cell line treated with the fermentation composition of the present invention.
Figure 5 is a photograph showing Chromatin staining and apoptosis by Hoechest 33342;
Fig. 6 shows the results of gelatin zymography showing the effect of reducing cancer metastasis.
FIG. 7 is a ROS staining photograph showing that a large amount of ROS is generated in mitochondria and cancer cells are killed.
8 is an immunoblot analysis showing the change in intracellular protein treated with a crude sample.
9 shows (a) photographs of kimchi according to the concentration and fermentation days of the fermentation composition and (b) MTT assay for colon cancer cell line HCT-116.
10 is a graph showing IC ₅₀ values and changes of colorectal cancer according to the concentration of the fermentation composition and the number of days of fermentation.
11 to 19 are photographs showing the visual change of the kimchi for 0 to 8 days, respectively.

Specific features and advantages of the present invention will be described in detail below with reference to the accompanying drawings. The detailed description of the functions and configurations of the present invention will be omitted if it is determined that the gist of the present invention may be unnecessarily blurred.

The present invention relates to a kimchi containing a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi and a preparation method thereof, and more particularly to a kimchi containing lactobacillus planta DGK-17 KACC 81028BP) was used to prevent the deterioration of the quality of kimchi caused by the administration of lactic acid-producing bacteria as well as the antimicrobial activity against the colonic cancer and antimicrobial activity against food poisoning bacteria at low concentrations. .

A method for producing a kimchi comprising a fermented composition prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi according to the present invention comprises culturing Lactobacillus plantarum DGK-17 (KACC 81028BP) A fermentation composition preparation step (S100) for preparing a composition, and a kimchi preparation step (S200) for immersing the kimchi in the fermented composition.

The fermentation composition preparation step (S100) is a step of preparing a fermented composition for inputting and mixing in the preparation of kimchi, which comprises culturing Lactobacillus planta DGK-17 (KACC 81028BP) And a concentration step for formulation.

More specifically, the fermentation composition preparation step (S100) comprises a seed culture step (S110) of producing a seed culture by sown-culturing Lactobacillus plantarum DGK-17 (KACC 81028BP) in an MRS medium and a seed culture step (V / v) to a medium containing 5 to 15 (w / v)% of at least one of the powder, the pulp, the pulverulent powder and the fruit powder at 150 to 300 rpm, (S120) for culturing the present culture for 24 hours, and a concentration step (S130) for concentrating the present culture.

The culture medium for culturing Lactobacillus plantarum DGK-17 (KACC 81028BP) in the fermentation composition preparation step (S100) is not limited as long as it provides nutrients necessary for microorganism growth, but is more preferably economical, It is also possible to use a complex natural medium which is easy to use, or to use a mixture of a complex natural medium and a synthetic medium.

For example, the composite natural medium may include one or more of cereal powder, soybean flour, and fruit powder, wherein the cereal is rice, glutinous rice, brown rice, wheat, barley, corn, oats, Mung beans, kidney beans, peas, peanuts, and the above fruits may include, but are not limited to, grapes, citrus fruits, oranges, and tomatoes. More preferably, soybean powder rich in nutrients is used.

In the fermentation composition preparation step (S100), it is preferable to autoclave the medium to increase the culture efficiency before culturing.

The concentration step (S130) is a step of preparing and concentrating the culture liquid, and the culture liquid can be concentrated by using lyophilization or concentration under reduced pressure at a low temperature.

The concentration step (S130) comprises a centrifugation step (S131) of centrifuging the present culture at 8,000 to 12,000 rpm at 4 DEG C for 5 to 15 minutes to recover the supernatant, and a pH adjustment step of adjusting the pH of the supernatant to 6.5 to 7.5 Step S132 and a formulation step S133 of formulating a pH-adjusted supernatant.

In the centrifugation step (S131), the culture medium is centrifuged at 4,000C for 5 minutes to 15 minutes at 8,000-12,000 rpm, and the separation efficiency is high at the rotation speed, time and temperature.

In the pH control step (S132), the pH of the supernatant is adjusted to 6.5 to 10.0, in consideration of the characteristics of the lactic acid bacterium having weak acidity and activity under the weak alkaline condition, during freeze drying or low- 7.5.

The formulation step (S133) may include a freeze-drying step (S133-1) of lyophilizing the pH-adjusted supernatant by pulverization or a low-temperature decompression step (S133-2) of concentrating the pH-adjusted supernatant under reduced pressure at 10 to 50 ° C You can do either one.

Conventional methods and conditions can be used for lyophilization. In the case of concentration under reduced pressure, the lyophilization is carried out at a relatively low temperature of 10 to 50 DEG C, whereby the death and disappearance of the lactic acid bacteria can be prevented at the temperature condition.

That is, the pH-adjusted supernatant through the pH-controlling step (S132) may be formed into a powdery form through a lyophilization step (S133-1) or through a low-temperature depressurization-concentrating step (S133-2) to remove the decompressed concentrate or the reduced- Can be formulated in the form of a dried powder.

In the present invention, the term " kimchi " refers generally to those generally known in the art which are prepared by adding seasonings to the main ingredients of ordinary kimchi.

The main ingredients of the kimchi may vary depending on the kind of kimchi to be produced. For example, the main ingredients of kimchi may be Chinese cabbage, radish, mustard, or heat.

In addition, the seasonings may include materials such as wave, garlic, ginger, liquid milk, sugar, salt, red pepper powder and the like. These materials may also vary depending on the kind of kimchi to be manufactured and the main ingredients.

The kimchi prepared by adding seasonings to the main ingredients may be, for example, Chinese cabbage kimchi, mustard kimchi, non-kimchi, kakdugi, etc. However, the present invention does not necessarily limit the kinds of kimchi. That is, the step of producing the kimchi according to the present invention may be one which is produced by a method generally known in the art.

In the present invention, a kimchi is prepared using the Chinese cabbage kimchi as an example, and the kimchi preparation step (S200) of the present invention for producing a Chinese cabbage kimchi comprises preparing a pickled cabbage (S210), a seasoning preparation (S220), a fermentation step (S230) of fermenting the seasoned kimchi, and a fermentation step (S240) of fermenting the fermented kimchi.

The prepared Chinese cabbage preparation step (S210) is a preparation step (S211) for cutting the Chinese cabbage after cutting the outer lobes and shims (S211), a washing step (S212) for washing the selected Chinese cabbages, (S213) for washing the pickled Chinese cabbage one to five times (S214), and a dehydration step (S215) for dehydrating for 30 minutes to 3 hours after the washed pickled Chinese cabbages are loaded on the dehydrated container But are not limited thereto.

The seasoning preparation step (S220) is a step of mixing the fermented composition and any seasonings.

In this case, the raw materials included in the above-mentioned seasonings are not limited, and may include base raw materials such as red pepper powder, green tea, sugar, and anchovy sauce, and solid raw materials such as rice wine, garlic, ginger, onion, May be manufactured by a first stage producing step (S221) of mixing the base raw materials first, and a secondary season preparing step (S222) of mixing the solid raw materials with the prepared base raw materials.

Since the fermentation composition is mixed with the solid raw material in mixing the fermented composition with any of the seasonings, the fermentation composition is introduced into the step of mixing the base material, that is, the primary seasoning step (S221) Can be mixed.

In the above-mentioned in-putting step (S230), the sauce is put on the pickled cabbage, and the sauteed sauce is filled and sealed in the package after fermentation, or filled and sealed before fermentation to store and deliver.

The fermentation step (S240) is a step of fermenting the kimchi seasoned with sauce, and may be fermented at 5 to 25 캜 in consideration of the time of ingestion of kimchi, production of lactic acid bacteria, and the like, but is not limited thereto.

For example, the kimchi preparation step (S200) of the present invention comprises 60 to 70% of Chinese cabbage, 8 to 10% of iris, 2 to 4% of anchovy sauce, 2 to 4% of garlic, 0.3 to 1% of ginger 1 to 3% of onion, 1 to 3% of onion, 3 to 5% of glutinous rice pulp, 0.5 to 1.5% of xanthan gum, 1 to 2% of white sugar, 0.1 to 0.5% of glutinous rice, 0.05 to 0.5% % Of leek, 1.5 to 3% of leek, and 0.05 to 10% of the fermented composition.

More specifically, the fermentation composition of the present invention may be contained in an amount of 0.05 to 10% by weight based on 100% by weight of the whole kimchi. If the content is less than 0.05% by weight, the effect of adding the fermentation composition is difficult to achieve. The flavor inherent to kimchi may be deteriorated, or corruption and odor may occur.

Hereinafter, the kimchi containing the fermented composition prepared from the novel lactic acid bacteria Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi according to the present invention is characterized in that it is produced by the above-mentioned production method.

The fermentation composition is prepared by adding 0.5 to 2% of a seed culture of Lactobacillus plantarum DGK-17 (KACC 81028BP) to a medium containing 5 to 15 (w / v)% of at least one of cereal powder, (v / v) and cultured at 150 to 300 rpm at 30 to 40 DEG C for 12 to 24 hours to prepare a culture solution, and concentrating the culture solution.

The fermentation composition is concentrated by centrifuging the present culture at 8,000 to 12,000 rpm at 4 ° C for 5 to 15 minutes to recover the supernatant, adjusting the pH of the supernatant to 6.5 to 7.5, lyophilization or heating at 10 to 50 ° C Followed by low-temperature decompression.

The kimchi containing the fermented composition is prepared by adding 0.5 to 10% by weight of the fermented composition among 100% by weight of the whole kimchi. The kimchi may have excellent taste and functionality in the weight range.

The fermentation composition has anticancer activity and antibiotic activity against food poisoning bacteria at low concentrations of 50 μg / mL to 100 μg / mL. More specifically, the fermented composition has an anticancer activity against HCT116 colon cancer cell lines. Pseudomonas aeruginosa 03K711 And Klebsiella pneumoniae 05K279.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1) Used strain and medium

Lactobacillus plantarum DGK-17 (hereinafter abbreviated as DGK-17) (hereinafter abbreviated as DGK-17) used in the present invention is a strain which is isolated and identified from kimchi and stored at the functional food research institute of Daegu University. Has antimicrobial effect against food poisoning bacteria and anticancer effect against colorectal cancer. In addition, the above-mentioned lactic acid bacteria strain has been deposited with the National Institute of Agricultural Science and is permanently preserved under the accession number KACC 81028BP on Oct. 11,

2) Preparation of fermentation composition

As an example of preparing the fermented composition using the above DGK-17, soybean powder was finely ground and diluted with distilled water to prepare a soybean flour liquid having a concentration of 10% and autoclaved.

The DGK-17 was seeded on MRS for 12 hours, and the culture was inoculated into the 10% soybean flour liquid solution autoclaved at 1% (v / v) and cultured at 200 rpm and 37 ° C for 16 hours to prepare a culture solution .

The culture solution was centrifuged at 10,000 rpm for 10 minutes at 4 ° C to separate precipitates. The supernatant was removed, pH was adjusted to pH 6.5 to 7 using 1M NaOH and lyophilized to obtain a powdered fermented composition. The fermented composition was stored at -20 < 0 > C.

3) Antimicrobial activity of fermented composition against food poisoning bacteria

3-1) Selection of target microorganisms

Two clinical isolates were P. aeruginosa 03K711, K. pneumoniae 05K279 and E. coli (Human) as food-borne pathogens, S. typhimurium ATCC 14028, Bacillus subtilis KCTC 6633, Bacillus cereus ATCC 14579, L. monocytogenes ATCC 19116, ATCC 19118, ATCC 19111 and ATCC 19166), and S. aureus KCTC1916 as a skin pathogen was prepared to confirm the antimicrobial activity.

As shown in Table 1, the growth of the microorganisms showed inhibition in Mueller Hinton Agar (MHA medium) prepared from the supernatant of DGK-17, and the antimicrobial activity of DGK-17 was inhibited by P. aeruginosa 03K711 and K. pneumoniae 05K279 , And the study focused on the above bacteria.

3-2) DGK With a pH of -17 In the bile salt  Resistance (Resistance)

Resistance (tolerance) to pH and bile acid salts was determined to show if it could withstand the intestinal environment.

Table 2 shows the resistance to acid and bile salt of DGK-17 exposed at 37 ° C for 4 hours at low pH and high bile salt concentration.

At pH 3, it showed higher resistance than pH 2. The bile salt resistance decreased with increasing bile acid concentration, but it showed that DGK-17 had sufficient resistance even at low pH and high bile salt content.

3-3) Antibiotic susceptibilities

Table 3 shows the results of the Antibiotic Susceptibility test of DGK-17. {R (resistant) is susceptible, S (susceptible) is susceptible.}

DGK-17 was shown to be susceptible to the glycolpeptide vancomycin, while aminoglycoside family streptomycin, antifungal amphotericin B, miconazole, cyclic polypeptide bacitracin, polyketide antibiotic tetracycline and b-lactam antibiotic Ampicillin.

3-4) For selected target microorganisms of fermented composition Inhibition

In order to show the inhibitory effect of the fermentation composition on P. aeruginosa 03K711 and K. pneumoniae 05K279, the A ₆₀₀ was measured three times and averaged.

Figure 1 shows a graph depicting the inhibition of selected target microorganisms of a fermentation composition according to an embodiment of the present invention.

Addition of the fermentation composition in the initial growth phase resulted in a decrease in the growth of P. aeruginosa 03K711 and K. pneumoniae 05K279, but increased in the absence of the fermentation composition.

3-5) Cell morphology of target microorganisms treated with fermentation composition

Fig. 2 is a SEM photograph showing changes in cell morphology when treated with P. aeruginosa 03K711 and K. pneumoniae 05K279 of the fermentation composition.

A shows a non-treated group K. pneumoniae 03K711 showing a smooth shape and B showing irregular shape showing K. pneumoniae 03K711 treated with a fermented composition.

C shows a non-treated group P. aeruginosa 05K279 showing a smooth shape, and D shows an irregular, twisted shape showing P. aeruginosa 05K279 treated with a fermented composition.

From the above results, it was confirmed that when the fermentation composition was added, cell membrane lysis of P. aeruginosa 03K711 and K. pneumoniae 05K279 was induced.

3-6) Auto-aggregation and co-aggregation

Figure 3 shows the ability of DGK-17 to colonize microorganisms, measured by autoaggregation and co-aggregation.

Statistical significance was measured by Student's t-test (* p <0.05, ** p <0.01, *** p <0.001).

DGK-17 showed a tendency to coaggregate more co-aggregate with K. pneumoniae 05K279 than P. aeruginosa 03K711.

4) Anticancer effect of fermented composition on colorectal cancer

4-1) MTT  assay

Figure 4 shows the MTT assay of HCT 1116 cell, a colon cancer cell line treated with the fermentation composition of the present invention.

As a control, oxaliplatin (25 μM), a drug used for colorectal cancer treatment, was used. In the MTT assay treated with a crude sample, an IC ₅₀ value of 100 μg was observed for HCT 1116 cells, a colon cancer cell line.

(4-2) Hoechest  Chromatin staining by 33342 and apoptosis  observe

The crude sample was treated with Hoechest 33342 stain to observe the effect of apoptosis. The result was similar to that of oxaliplatin, the control group. These results also demonstrate a strong anti-cancer effect. (Shown in Fig. 5)

(4-3) Gelatin zymography  Reduction of cancer metastasis through

Gelatinase activity of the crude sample was measured and the amounts of MMP complexes and MMP-9, Pro-MMP-2 and MMP-2 were decreased compared to the control. This result shows that the fermentation composition has a reduced effect of darkening (see FIG. 6).

(4-4) ROS  staining

As a result of evaluating reactive oxygen species (ROS) staining for active oxygen production by treating a crude sample, a large amount of ROS was produced in the mitochondria and the cancer cells were killed (see FIG. 7).

(4-5) Western blotting

A crude sample was processed to analyze the changes in intracellular proteins by immunoblot analysis. Caspase 3, Cytochrome C (Cyt. C), Bcl2, and Bax, which are markers of apoptosis pathway related to apoptosis, were observed.

As a result, expression of Caspase 3, Cytochrome C (Cyt. C) and Bax was increased, and Bcl2 was decreased. Thus, apoptosis was observed (as shown in FIG. 8)

5) Manufacture of Kimchi

Were mixed at a blending ratio according to the following Table 4 to prepare a kimchi. In Comparative Example 1, the fermentation composition was not added. In Examples 1 to 3, 7.5 g, 15 g, and 30 g of the fermentation composition were added per 1 kg of the kimchi.

6) Depending on the concentration of fermented composition added to kimchi MTT  assay

The kimchi was prepared according to Example 1 of Table 4, except that the concentrations of the fermentation composition were varied to 50, 100 and 200 ㎍ / mL, respectively, and MTT assay was performed according to the concentration of the fermentation composition.

9 shows (a) photographs of kimchi according to the concentration and fermentation days in the fermented composition and (b) MTT assay for colon cancer cell line HCT-116 (@@@ P <0.001, <0.05 vs Control)

The survival rate of colorectal cancer cell line HCT-116 after 5 days of fermentation at 25 ° C was significantly decreased after 5 days of fermentation, and the survival rate of colon cancer cell line HCT-116 was remarkably decreased. And decreased cell survival rate.

As a result, it was confirmed that the fermentation composition of the present invention had an excellent anti-cancer effect even at a low concentration of 50 μg / mL.

7) IC according to the addition amount of fermented composition added to kimchi ₅₀ Change in value

The kimchi was prepared according to Example 1 of Table 4, except that the concentration of the fermentation composition was changed to 50, 100 and 200 ㎍ / mL, respectively, and then the IC ₅₀ value and the change according to the concentration and fermentation days of the fermentation composition were determined Respectively.

Table 5 shows IC ₅₀ values and changes according to the concentration of the fermentation composition and the number of days of fermentation, and FIG. 10 is a graphical representation thereof.

Overall, the higher the concentration of the fermentation composition, the lower the IC ₅₀ value, and the decrease with the flow of fermentation days decreased from 0 to 5 days, but there was no significant change thereafter.

8) pH change according to fermentation days of Kimchi

Table 6 shows the pH changes of kimchi prepared according to Table 4 according to the days of fermentation.

It was difficult to ascertain the increase or decrease of pH depending on the number of days of fermentation, and it was confirmed that it was kept constant as a whole.

9) Sensory evaluation

The sensory evaluation of the kimchi prepared according to Table 4 was carried out for a total of 20 persons, each of 10 skilled persons in their 20s. The evaluation was carried out according to the 5 point scaling method. In other words, the score was given as 1 point: very bad, 2 points: poor, 3 points: normal, 4 points: good, and 5 points: very good. The evaluation items were evaluated by taste, smell, color, refreshing feeling, texture, overall taste, and six items.

In Comparative Example 1, the fermentation composition was not added. In Examples 1 to 3, 7.5 g, 15 g, and 30 g of the fermentation composition were added per 1 kg of the kimchi. The kimchi was stored at 25 ℃ for 0 ~ 8 days and sensory evaluation was performed once a day.

Figs. 11 to 19 show the visual changes of the kimchi for 0 to 8 days, respectively.

Table 7 shows the sensory evaluation results of the taste.

Table 8 shows the sensory evaluation results for odor.

Table 9 shows the sensory evaluation results of the sensation of refreshing.

Table 10 shows the results of sensory evaluation for color.

Table 11 shows the results of sensory evaluation for texture.

Table 12 shows the sensory evaluation results for the overall preference.

As a result of analyzing sensory evaluation results, taste, smell, refreshing feeling, color, texture, overall taste were not significantly different in all items. However, the taste, smell, refreshing sensation, and overall acceptability in the detailed items showed good sensory evaluation results in the order of Sample 3> Sample 2> Sample 1> untreated. On the contrary, color and texture were the opposite in the order of sample 3 <sample 2 <sample 1 <control untreated. These results suggest that the inoculation effect of lactic acid bacteria Lactobacillus plantarum DGK-17 contained in the fermentation composition may have caused some influence on the ripening of kimchi.

In addition, it was confirmed from the sensory evaluation results that the taste and the quality of the kimchi were not deteriorated by the addition of the lactic acid bacteria and the fermented composition.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken as limiting the scope of the present invention. The present invention can be variously modified or modified. The scope of the invention should, therefore, be construed in light of the claims set forth to cover many of such variations.

National Academy of Agricultural Sciences KACC81028BP 20161011

Claims (14)

delete delete delete delete delete delete delete delete A method for producing a kimchi comprising a fermented product produced from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi,
A fermentation product preparation step (SlOO) for culturing Lactobacillus planta DGK-17 (KACC 81028BP) to produce a fermented product;
And a kimchi preparation step (S200) of immersing the kimchi into the fermented product,
The fermentation product preparation step (SlOO)
A seed culture step (S110) in which a Lactobacillus planta DGK-17 (KACC 81028BP) is seeded in an MRS medium to produce a seed culture liquid;
The seed culture was inoculated in a medium containing 5 to 15% (w / v)% of soybean powder so as to be in a concentration of 0.5 to 2% (v / v) and cultured at 150 to 300 rpm at 30 to 40 ° C for 12 to 24 hours. (S120), which comprises:
(S130) of concentrating the present culture liquid,
The concentration step (S130)
A centrifuging step (S131) of centrifuging the present culture at 8,000 to 12,000 rpm at 4 DEG C for 5 minutes to 15 minutes to recover the supernatant;
Adjusting the pH of the supernatant to 6.5 to 7.5 (S132); and
and a formulation step (S133) of formulating a pH-adjusted supernatant,
The formulation step (S133)
(S133-1) in which the pH-adjusted supernatant is lyophilized and pulverized, or the low-temperature reduced-pressure concentration step (S133-2) in which the pH-adjusted supernatant is concentrated under reduced pressure at 10 to 50 ° C,
The kimchi
HCT116 colon cancer cell line and has an antimicrobial activity against any one or more of Pseudomonas aeruginosa 03K711 and Klebsiella pneumoniae 05K279
A method for producing a kimchi comprising a fermented product prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 derived from kimchi.
delete delete delete 10. The method of claim 9,
The kimchi manufacturing step (S200)
Preparation of pickled cabbage (S210); and
A seasoning preparation step (S220) of mixing the fermented product and any seasonings;
A step S230 of filling the sauce made with pickled Chinese cabbage;
And a fermentation step (S240) of fermenting the kimchi mixed with the sauce
A method for producing a kimchi comprising a fermented product prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 derived from kimchi.
14. The method of claim 13,
The seasoning preparation step (S220)
Characterized in that 0.5 to 10% by weight of the fermentation product in 100% by weight of the whole Kimchi is mixed
A method for producing a kimchi comprising a fermented product prepared from a novel lactic acid bacterium Lactobacillus plantarum DGK-17 derived from kimchi.

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