KR100434450B1 - Anticancerous green onion kimchi and process for preparation thereof - Google Patents

Abstract

The present invention relates to an anti-cancer functional kimchi and a method of manufacturing the same. The present invention, kimchi, which is soaked in saline instead of anchovy, has excellent taste as well as antimutagenic and anticancer effects. Sesame kimchi has an excellent anti-cancer effect. In addition, the present invention sake kimchi is packed in a flexible aluminum wrapper with a gas absorbent and stored at 10 ° C for excellent preservation. This is a very useful invention in the food industry.

Description

Anti-cancer functional green kimchi and manufacturing method thereof

The present invention relates to anti-cancer functional kimchi and a method of manufacturing the same. More specifically, the present invention relates to a high-preservation, good taste, excellent anti-cancer effect, and a method for producing the kimchi prepared according to the standardized recipe for preparing kimchi after pickling leek in saline.

The leek is colder than the leek, so it can produce high quality leek in the place where sunlight can be received a lot in the winter. Due to the characteristics of these leeks, kimchi is mainly favored for cultivation of leek, and in the southern part of Gyeongsang-do, it is mixed with rich anchovy fish sauce in spring and autumn to enhance the appetite. On the contrary, due to the rich flavor of the sauce and the unique aroma of green onions, Pa Kimchi is not available in the popular Kimchi-ro, which is enjoyed by anyone like cabbage kimchi and kakdugi. , Park Gun Young: Local specialties Kimchi and its characteristics in Busan, Gyeongnam. Korean Journal of Nutrition and Nutrition, 24 (5), 41-49 (1995), Han Sook-Hee, Lee Sook-Hee, Kyung-Im Lee, and Kun-Young Park: Standardization of Kimchi, Gyeongsang-do. Journal of the East Asian Society of Dietary Life, 5 (2), 32-40 (1995)]. Pusan National University Master's Thesis (1999)] is just beginning.

As Kimchi, Korea's most representative food, is becoming known throughout the world, antimutagenic effects caused by lactic acid bacteria produced by fermentation with vitamin C, β-carotene and flavonoids in kimchi, Anticancer, immune enhancing effect [Hertog, MGL, Hollman, PCH and Katan, MB ： Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands. J. Agric. Food Chem., 40, 2379-2383 (1992), Kyung-Im Lee, Gun-Young Park: Anti-cancer effect of green and yellow vegetables. Journal of Life Science, 3, 143-152 (1993), Sun Mi Lee: Antimutagenic and Anticancer Effects of Kale and Its Mechanism. Pusan National University PhD Thesis (1995), Kazuki, S., Seikichi, K .., Misao, M., Zwe-Ling, K., and Hiroshi, H .: Antimutagenicity of dialyzates of vegetables and fruits. Agric. Biol. Chem., 52 (6) 1369-1375 (1988)] and its functional excellence and anti-mutation and antioxidant effects of vegetables used as ingredients of kimchi such as red pepper powder and garlic [Lee Jun-young, Lee Na-young, Lee Hyun-cheol, Jung Seon-sik] : Effect of retinoic acid, β-carotene and ascorbic acid on the mutagenesis of anticancer antibiotics in the Ames test. Journal of the Korean Cancer Society, 24, 504-515 (1992), Park Gun-young, So-hee Kim, Signer, Hae-Young Jung: Garment Mutation Inhibition and Growth Inhibitory Effect of HT-29 Colon Cancer Cells. Korean Journal of Food Science and Technology, 23, 370-374 (1991), Eun-Sil Kim, Hee-Jung Chun: A Study on the Anticancer Effect of Garlic on DMBA Carcinogenicity in Hempster. Korean Journal of Nutrition and Nutrition, 22 (4), 398-404 (1993), Ryu, SH, Jeon, YS, Kwon, MJ, Moon, JW, Lee, YS and Moon GS: Effect of kimchi extracts to reactive oxygen species in skin cell cytotoxicity. J. Korean Soc. Food Nutr. , 26, 814-821 (1997), Mun, SI, Ry, HS, Lee, HJ and Choi, JS: Further screening for antioxidant activity of vegetable plants and its active priniciples from Zanthoxylum schimifolim. J. Korean Soc. Food Nutr. , 23, 466-471 (1994), Seung Hoo Lim, Tae Hyo Kim: In vitro Biological Activity of Allicin and Ethanol Extracts from Korean Garlic. Korean Journal of Food Science and Technology, 29 (2), 348-354 (1997), Kun-Young Park, Hong-Sik Choi: Antimutagenicity and Anticancer Activity of Kimchi. Paper presented at the 1 st Kimchi Symposium. Korean Food Science Society, 205-225 (1994)] is also known and studied a lot. Most of these studies on kimchi are the functional research on the most common cabbage kimchi, and there is almost no research on the functionality of leek itself as well as green kimchi.

The inventors of the present invention reduced the amount of fish sauce and reduced the amount of fish sauce so that the kimchi is not only local kimchi that is limited to the southern region, but also produced the kimchi that uses salt and fish sauce together. Based on the taste of the food, ingredients were selected through physicochemical experiments and sensory tests to improve the taste and preservation. In addition, to determine the antimutagenicity and anticancer function of pakimchi kimchi through the in vitro experiments and to confirm these functions in vivo in vivo chemokimchi administered to the Balb / c mice transplanted sarcoma-180 tumor cells to change the weight of each organ The present invention by examining the activity change and glutathione content change of glutathione S-transferase (GST: nmol protein / min), a phase II enzyme present in the liver, the effect of inhibiting solid cancer growth, NK cell immune activity Was completed.

Accordingly, it is an object of the present invention to provide a method of preparing green onion kimchi with excellent taste and preservation while anti-cancer by slicing green onions with brine having different kinds of salts and mixing subsidiary materials.

Another object of the present invention to provide a kimchi prepared by the above method.

The above object of the present invention is the physicochemical properties of each green kimchi prepared by marinating green onions in salted and salted anchovy sauce mixed with red pepper and anchovy marinated and seasoned with anchovy sauce mixed with red pepper powder. And the sensory test to confirm that the sauerkraut prepared by pickling in saline has excellent flavor, and the sauerkraut prepared by the saline pickling method in various packing papers and stored at various temperatures to investigate the physicochemical properties of the sauerkraut. And packaging containers were examined. For the sensory test, the saline pickled green kimchi, the pickled green kimchi, and the cabbage kimchi extract of general cabbage kimchi were investigated for antimutagenicity and anticancer activity in vitro , and the saline pickled green kimchi showed a significantly high anticancer effect. In vivo micronucleus test, solid cancer growth inhibition test, spleen NK cell activity test, activity of glutathione S-transferase (GST: nmol protein / min), a phase II enzyme present in liver, and Glutathione content change experiment was also achieved by obtaining the same results as in vitro experiments.

Hereinafter, the configuration of the present invention will be described.

Figure 1 shows the results of measuring the change in pH and acidity while fermenting the kimchi pickled and brine pickled green kimchi at 15 ℃ each.

Figure 2 shows the results of measuring the change of Lactobacillus sp. ( Lactobacillus sp. ) Strains and Leuconostoc sp. Strain while fermenting each of the anchovy pickled and brine pickled green kimchi at 15 ℃.

Figure 3 shows the results of the sensory test of each of the anchovy pickled and salted green kimchi.

Figure 4 shows the results of measuring the change in pH, acidity while fermenting kimchi at various storage temperatures.

Figure 5 shows the results of measuring the change of Lactobacillus sp. ( Lactobacillus sp. ) Strain and Leuconostoc sp. Strain while fermenting the kimchi at various storage temperatures.

Figure 6 shows the results of measuring the change in pH and acidity while fermenting kimchi wrapped in vinyl and aluminum 10 ℃.

Figure 7 shows the results of measuring the volume expansion change while packing the kimchi with aluminum or aluminum with a gas absorber and fermentation at 10 ℃.

Figure 8 shows the results of measuring the pH and acidity while packing kimchi into aluminum, aluminum and plastic bottles with a gas absorbent, respectively, and fermentation at 10 ℃.

9 shows oral administration of pakimchi methanol extract to mice and after injection of mitomycin C, a micronucleus-inducing substance, counts micronucleus induction.

Figure 10 shows the results of measuring the activity of natural killer (NK) cells in the spleen of mice administered orally with the kimchi methanol extract.

Fig. 11 shows the effect of pakimchi methanol extract on the activity of glutathione-S-transferase in liver tissue of Balb / c mice transplanted with sarcoma-180 cells.

Figure 12 shows the effect of methanol extract of green kimchi on the glutathione content in liver tissue of Balb / c mice transplanted with sarcoma-180 cells.

The present invention comprises the steps of marinating green onions in anchovy or salted with saline, and mixing the subsidiary materials such as red pepper powder, salt at an appropriate ratio to produce fermented kimchi; Investigating the pH, acidity, and lactic acid bacteria of each of the prepared kimchi marinated green kimchi and sauerkraut pickled in saline to investigate the physicochemical properties and to perform a sensory test; Determining the optimum storage temperature by packaging the kimchi prepared by brine with the salt removed from the brine and examining the physicochemical properties of kimchi while storing them at various temperatures; Determining the optimum preservation wrapper by investigating physicochemical properties while storing the kimchi in various wrappers under the optimal preservation temperature; Investigation of antimutagenic effects by performing SOS chromotest in vitro on green kimchi salted with saline and different cultivation methods using different types of salted green kimchi salt. Investigating the anticancer effect of sauerkraut pickled with different kinds of salts in vitro by MTT assay; Investigating the anticancer effects of green kimchi and sauerkraut kimchi, which were salted and pickled in different types of salted green kimchi and salt, by MTT assay in vitro ; Different kinds of pickled green kimchi and salt were used as samples to feed the mouse, and micronucleus-induced substances were administered to the mice, and then the incidence of micronucleus in the reticulocytes of peripheral blood was investigated. step; After transplanting sarcoma-180 tumor cells to the mice, the salted green kimchi, anchovy sauerkraut and cabbage kimchi sample solution, which were salt-pickled with different salts, were fed to the mouse and the weight of solid cancer was measured. Confirming; Checking the activity of spontaneous killer cells in the spleen of the mouse, which measured the weight of the solid cancer, to confirm the effect of enhancing the immunological activity of green kimchi; Hepatic glutathione content and glutathione-S-transferase activity of the mouse measured the weight of the solid cancer consists of the step of confirming the immune activity enhancement effect of green kimchi.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1 Preparation of Green Kimchi

The onions used in this example were purchased at the Bujeon Market in Busan on the day of dipping into the traditional leek of Kijangsan. Chopped chives were chosen to be not thick in length but not thick in leaves, and not dried in leaves. Organic traditional onions were sold at Advent Natural Farm in Milyang, Gyeongnam. Namhae garlic and Chungbuk Seosan Ginger were purchased at the Busan Bujeon Market on the day of Kimchi. Also, red pepper powder is the most suitable product for color, sensory test, and flavored kimchi. It has red color and has little spicy taste. Anchovy sauce is a pure anchovy sauce with Ulsan Gangdong anchovy sauce. There was no smell, and the fish sauce was used. Glutinous rice paste is boiled and cooled with the ratio of glutinous rice flour and water at 1: 8, sesame seeds are made from China or Sudan, sugar is refined sugar, salt is baked salt [Sannae, Co., Ltd.], roasted salt [Wooil salt industry] Natural salts [Wild salt industry], which removed liver water, and general salts [Wild salt industry], which was not removed. However, in the pickling method, domestic salt from which salt was removed was used.

The mixing ratio of the seasoning used for the kimchi is the mixing ratio standardized by the present inventors and physicochemical experiments (Lim Ji-yeon 1999), and as shown in Table 1, the relative ratio with respect to the green onion 100.0 in the case of anchovy pickling method 20.0, red pepper powder 9.7 , Garlic 5.2, ginger 2.7, sesame seeds 1.3, sugar 1.5, glutinous rice grass 12.0, the other ingredients are the same as pickled salt, and the anchovy fish sauce was cut in half to 10.0 for chives 100.0.

In the picking with anchovy juice method, the soaking method and the fermentation temperature are not pickled with brine, but the white parts of the chives are immersed in anchovy sauce of an ingredient mix ratio at an angle and pickled for 2 hours at 4 ° C. Put green onion kimchi. In other words, the chives were finely rinsed, washed, and drained for 1 hour at 5 ° C. Then, they were spread out one by one, sprinkled with anchovy salt alternately, and then pickled at 5 ° C for 2 hours. At this time, the pickled roots were soaked in the roots so that they were well pickled and carefully turned over at about 30 minutes. When it is pickled, mix the red pepper powder with the salted anchovy sauce and mix it with the seasoning so that the remaining ingredients are soaked well. In picked with salted water, the chives were pickled in salted water at 4 ° C. for 30 minutes, and the amount of anchovy was reduced to half the amount of pickled fish. In other words, after trimming and washing the chives well, immerse the leek in 2% 5% brine of the leek weight and rinse for 30 minutes. At this time, the wave was pressed with a heavy one to completely immersed in saline. Pepper powder was mixed with anchovy sauce in advance, and when the chives were marinated, the water was taken out and then the onions were drained. After tossing, 5-8 strands of green onions were wrapped up or packaged as it was for fermentation.

Blending ratio of green kimchi samples Diversity Compound Pickled with anchovy sauce Pickled in brine wave 100.0 100.0 (pickles) chili powder 9.7 9.7 Anchovy liquid milk 20.0 10.0 chopped garlic 5.2 5.2 Chopped ginger 2.7 2.7 Sesame 1.3 1.3 Sugar 1.5 1.5 Glutinous rice paste 12.0 12.0 Final salt concentration 2.3% 2.3%

Example 2: Physicochemical Properties of Kimchi Prepared by Different Pickling Methods

In the present Example, the pH, acidity, and lactic acid bacteria were measured by storing the brine pickled green kimchi and anchovy pickled green kimchi prepared in Example 1 at 15 ° C., and the fermentation patterns were examined.

The pH was measured at room temperature with a pH meter (Corning 220, USA), and the acidity was diluted 20 times with 20 mL of the sample, and 10 mL of the sample was added. According to the AOAC method, 1 mL of 0.1% phenolphthalein was added as an indicator and 0.1 N NaOH. It was set as the end point by titrating with pink. The titration value was converted into lactic acid and expressed as% of content.

The plate count technique was used to measure the number of lactic acid bacteria. Dilute 1 mL of the sample solution with sterilized distilled water, dissolve 0.1 mL of each dilution solution in advance, dissolve it in 10 mL of MRS medium cooled to 43 ° C to 45 ° C, mix, make a plate in a petri dish, and make 37 ° C. The number of colonies shown by culturing in an incubator was counted and measured by the number of lactic acid bacteria. Lactobacillus bacteria (Lactobacillus) medium Lactobacillus selective medium (Lactobacillus selection medium; LBS medium) acetic acid (acetic acid) to suppress the growth of Phedi OKO kusu (Pediococcus) in the sodium acetate (sodium acetate) the deformation addition LBS Plates were incubated at 30 ° C. for 3 days using agar medium (modified LBS agar medium; m-LBS medium) (Table 2). Using phenylethyl alcohol sucrose agar medium (PES medium) added with phenylethyl alcohol and sucrose as leuconostoc selective medium (Table 3) Plates were incubated for 5 days at < RTI ID = 0.0 >

The sensory test was performed by eight sensory personnel trained according to a replicated randomized complete block design and evaluated three samples at a time and repeated four times. Descriptive items were subjective evaluations. Appearance, comprehensive evaluation, and objective evaluation items were taste perceptions such as sour taste, gundeok taste, unripe taste, salty taste, spicy taste, and toughness. The degree of appearance and comprehensive evaluation is not good when it is close to 1, but is excellent when it is close to 9, and the sour, salty, spicy, Gundeok, less-ripe, and chewy are weaker when it is closer to 1 and stronger when it is closer to 9. Indicated. Taste perception was assessed by the mouth and nose after chewing with molars several times, and the degree of toughness was evaluated by the strength of chewing two or three times in the same direction as the fiber using the incisor. At this time, the sour taste is defined as acid sense, the unripe taste is reminiscent of bitterness or astringent from green grass, and Gundeok taste is unpleasant to old kimchi.

As a result, as shown in FIG. 1, the pH of the salted pickled green kimchi was pH 4.2 and the salted pickled green kimchi was slightly higher at pH 15, and the salted pickled green kimchi was pH 4.3 at 10 ° C. . In case of acidity, the pickled green kimchi was 1.1% and the brine pickled kimchi was 1.13% at 10 days. Among the lactic acid bacteria, Leuconostoc sp. Strain, which is related to taste and gives a cool taste, was pickled at 2.5 × 10 ⁸ CFU / mL at 1.2 × 10 ⁹ CFU / mL in saline pickled green kimchi on the 10th day of the optimum season. It grew a lot compared to the kimchi. On the other hand, Lactobacillus sp. ( Lactobacillus sp.) Involved in the rancidity of kimchi, it can be seen that the brine pickled smaller kimchi grown in the optimum stage (Fig. 2). As shown in Fig. 3, the sensory test of pickled green kimchi and pickled green kimchi had a higher spicy taste than pickled green kimchi, but showed similar scores in most items such as appearance, comprehensive evaluation, and salty taste.

According to the results of the present example, the pH and acidity of the brine pickled kimchi compared to the fish pickled kimchi is similar, and the strain of Leuconostoc sp. Time savings and economical advantages were found.

Composition of m-LBS medium for culturing Lactobacillus sp. Ingredient m-LBS medium (%) peptone 1.0 gravy 1.0 Yeast extract 0.5 Glucose 2.0 Twin 80 0.1 K ₂ HPO ₄ 0.2 Sodium acetate 0.5 Ammonium citrate 0.2 MgSO ₄ 7H ₂ O 0.02 Acetic acid 2.5mL / L Sodium acetate 35 g / L

Composition of PES medium for culturing Lactobacillus sp. Ingredient PES badge (g) Trypticase 5.0 Yeast extract 0.5 Sucrose 20.0 (NH ₄ ) ₂ SO ₄ 2.0 MgSO ₄ 7H ₂ O 0.244 KH ₂ PO ₄ 1.0 Baby 15.0 Phenylethyl alcohol 2.5 water 1000 mL pH 9.8

Example 3: Green Kimchi Preservation Method

First Process: Determination of Optimal Temperature

In this process, in order to find a suitable temperature for the distribution of green kimchi, the seaweeds prepared by the brine pickling method in Example 1 were first packed with 500g each in a flexible aluminum package without a gas absorbent, and then at 0 ° C. After storage for 24 hours, the temperature was shifted to 0 ° C., 5 ° C., 10 ° C., and 15 ° C., and the kimchi was taken at regular intervals on this day to investigate the physicochemical change. 0 ° C. was measured for 7 days, 5 ° C. for 5 days, 10 ° C. for 3 days, and 15 ° C. for 2 days. PH, acidity, and lactic acid bacteria were measured by physicochemical changes and the degree of volume expansion of the wrapping paper was measured. The physicochemical change was measured in the same manner as in Example 2.

As a result, as shown in FIG. 4, the pH and acidity change according to temperature were pH 4.24 for 10 days at 15 ° C., and dropped to pH 4.00 or less after 12 days. The acidity of Pa Kimchi was higher than other Kimchi, and it was 0.5% or 0.6% higher on the 0 or 2 days, and the acidity was 1.13% on the 10th. The pH change was large during the 4 to 12 days and gradually changed thereafter. The pH change during fermentation at 10 ° C. showed a large pH change from 15 days to 21 days and was optimally fermented to pH 4.2 at 20 days, and remained at pH 4.0 after 20 days, but dropped to pH 3.88 at 36 days. As with pH, the change in pH from 15 to 21 days was 1.16% at 20 days of maturity. Acidity change by storage period was slower than 15 ℃. In case of 5 ℃, the degree of change was small, unlike 15 ℃ and 10 ℃, and the pH was lowered to pH 4.95 at 45 days and maintained at pH 4 or higher until the last day 60 days. As with pH, the acidity was also slowly changed. The change in acidity was 0.2% for 60 days and 0.81% for 60 days. At 0 ° C., the pH did not drop below pH 5 until 80 days, and the acidity was almost unchanged as pH, so the acidity was maintained at 0.63% at 80 days.

In the loconocstock, a lactic acid bacterium that gives kimchi flavor and a cool taste.Leuconostocsp.) Lactobacillus genus (lactic acid bacterium)Lactobacillussp.) The result of measuring the strain is, when stored at 15 ℃, as shown in Fig.Leuconostocsp.) strain gradually increased and reached the maximum value at 8 to 10 days, but gradually decreased as it matured. Genus LactobacillusLactobacillussp.) strains decreased slightly from 8 to 10 days and then increased as they matured. In storage fermentation at 10 ℃,Leuconostocsp.) strain gradually increased and showed a maximum value between 18 and 27 days, but gradually decreased as it matured. Genus LactobacillusLactobacillussp.) strains were slightly decreased and then increased as ripening occurred. The appearance of pavement expansion began to expand from day 12 but was less than that of 15 ℃. At 5 ° CLeuconostocsp.) strains and the genus Lactobacillus (Lactobacillussp.) all strains gradually increased up to 60 days. When stored at 0 ℃Leuconostocsp.) strains of the genus Lactobacillus (Lactobacillussp.) more than strain It grew up.

As a result, for the commercial distribution of green kimchi, it was found that the lactic acid bacterium that grows a lot grows, and 10 ° C., which has a low pH and acidity change, is an appropriate storage temperature.

Second Process: Determining Optimal Wrapping Paper

In this process, in order to find a suitable wrapping paper for the distribution of green kimchi, the seaweeds prepared by the brine pickling method using the salt water removal salt in Example 1 were placed in various packaging papers at 10 ° C. determined at the optimum temperature when stored in the first step. The pH and acidity over time were examined in the same manner as in Example 2 while keeping for a day. At this time, the wrapping paper for packing the kimchi is a flexible wrapping paper, 500g capacity of general vinyl wrapping paper, 500g capacity of aluminum wrapping paper (PET / AL / PE) that is not treated with gas absorbent, and aluminum wrapping paper (PET / AL / PE) with gas absorbing agent. 500 g capacity was used. In addition, for comparison experiments, the packaging container was packed using a transparent plastic bottle (PET), packed with kimchi, and stored at 0 ° C. for 24 hours, and then moved to 10 ° C. to measure pH and acidity every four days. At this time, Ca (OH) ₂ was used as the gas absorbent of the flexible packaging paper. The volume expansion of the package was measured using a laboratory-made measuring cylinder, and the water was poured into the pouch to obtain the volume of the pouch from the increased volume of water.

As a result, as shown in FIG. 6, aluminum was pH 4.2, acidity was 1.16%, vinyl was pH 4.15, and acidity was 1.27% at 20 days of maturation. In addition, the degree of expansion of the wrapping paper volume was not significantly different at the beginning as shown in FIG. 7, but as the fermentation proceeded, the wrapping paper without the gas absorbent was expanded significantly compared with the wrapping paper with the gas absorbent. In addition, the results of comparison using aluminum wrapping paper, aluminum wrapping paper treated with a gas absorbent, and transparent plastic bottles (PET bottles) did not show a big difference for each packaging container during the fermentation process as shown in FIG. In the plastic bottle of kimchi, the pH dropped a little later, but it was similar to the group that attached gas absorbent to aluminum as the ripening machine passed. In the case of aluminum wrapping paper, the pH tended to fall faster between 16 and 20 days than the other groups. In case of acidity, aluminum wrapping paper increased the fastest and plastic bottles showed the most slowly increasing. As a packaging container for the kimchi distribution, aluminum wrapping paper with a gas absorbent and a transparent plastic bottle were found to be suitable. However, in the case of plastic bottles, but there is a shelf life, the seasoning is sagging down and the seasoning is separated and the texture is tough, there is a disadvantage that the seasoning can overflow when aged. Therefore, it was found that aluminum packaging paper having a gas absorbent is most suitable as a packaging container for the distribution of green kimchi.

Example 4: in vitro Antimutagenic Effects of Green Kimchi by SOS Chromotest

In this embodiment, green kimchi and salt baked with salted pickling method in Example 1, sea salt removed from salted water, green kimchi prepared with salted pickling using Hanjugeum, and standardized Chinese cabbage kimchi recipe [Jo Eun-ju: Standardization and cancer prevention effect of cabbage kimchi . Pusan National University Graduate School Ph.D. Thesis (1999)] was used to freeze-dried kimchi (pH 4.2) in the ripening stage, containing general cabbage kimchi, extracted with methanol, and antimutagenic in SOS chromotest using E.coli PQ 37 strain in vitro . The effect was found. In the case of grilled salt, the normal and organic chives were used to compare the functionality. That is, 50 µl of frozen PQ37 bacteria were inoculated into 5 mL L culture medium, shaken overnight at 37 ° C, and then inoculated again in 5 mL L culture medium, and shaken for 2 hours until the A660 measurement reached 0.3 to 0.4 at 37 ° C. It was. The obtained microbial solution was diluted 1/10 in L culture medium, and 100 μl of each well of a 96 well plate prepared by preliminarily dispensing 20 μl of the sample prepared by each concentration and the mutagen was mixed and shaken at 37 ° C. for 90 minutes. After inducing the SOS response, on one side, O-nitrophenyl-β-D_galactosyranoside (O-nitrophenyl-β-D- for measuring activity of β-galactosidase (β-G) 100 μl of galactopyranoside (ONPG) and 100 μl of P-nitrophenyl phosphate disodium (PNPP) were added to the other side to measure the activity of alkaline phosphatase (AP). The color development time was 30 minutes, β-G was 100M of 1.5M Na ₂ CO ₃ , AP was 50L of 1M HCl, and the color development reaction was stopped by enzyme. After 5 minutes, 50μL of 2M tris buffer was added to the AP side. HCl was added to neutralize and the absorbance was measured at 420 nm with a spectrophotometer. The measured OD 420nm measured enzyme value (Eu) by Miller's formula.

Eu = (1000 × A ₄₂₀ ) / t (min)

At this time, the experimental strain used in this example was E. coli PQ37 derived from E. coli GC4436 strain was mixed with 1: 1 bacteria cultured overnight in 90% glycerol (L) and L medium (L medium) Store at -20 ° C. The strains were prepared every six months, and each time the uvrA mutation, rfa mutation and PHOc gene constitutivity and sfiA :: lacZ fusion were tested. Mutagenesis N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was purchased from Aldrich, USA and dissolved in distilled water.

As a result, as shown in Table 4, green kimchi with roasted salt showed the highest inhibitory effect of 70% at the concentration of 50㎍ / plate. There was a difference between organic and normal kimchi at 60% of organic kimchi and 50% of normal kimchi at low concentration, but there was no difference at high concentration. Sea salted kimchi with brine showed slightly lower inhibition rate than roasted salt. In the case of Hanju Geumpa Kimchi, it showed the lowest effect among the Kimchi groups, and the pickling leek kimchi showed lower effect than the Korean cabbage kimchi at low concentration, but higher effect than Chinese cabbage kimchi at high concentration. The general cabbage kimchi was also contained as a kimchi removed from the liver, so the kimchi was more effective when comparing the antimutagenic effects of cabbage kimchi and green kimchi.

SOS Inhibitory Effects of Methanol Extracts from Green Kimchi on MNNG (70ng / assay) in E.coli PQ37 Strains Sample β-galactosidase (β) Alkaline phosphatase (ρ) β / ρ SOS Inducer Inhibition Ratio (%) OD ₄₂₀ Unit OD ₄₂₀ Unit Voluntary 0.321 ± 0.011 10.69 0.346 ± 0.010 11.54 0.93 1.00 Control 1.013 ± 0.036 33.78 0.332 ± 0.004 11.08 3.05 3.29 25 µg / plate Gs ¹⁾ OGK 0.582 ± 0.051 19.56 0.330 ± 0.007 11.00 1.78 1.92 60 Gs GK 0.612 ± 0.089 20.39 0.308 ± 0.008 10.28 1.98 2.14 50 BRCs ²⁾ GK 0.664 ± 0.016 22.14 0.350 ± 0.012 11.67 1.90 2.05 54 Hs ³⁾ GK 0.874 ± 0.016 29.13 0.372 ± 0.014 12.40 2.35 2.54 33 Aj ⁴⁾ GK 0.781 ± 0.096 26.04 0.351 ± 0.019 11.71 2.22 2.40 39 KcK ⁵⁾ 0.764 ± 0.010 0.36 0.357 ± 0.016 11.91 2.14 2.31 43 50 µg / plate Gs OGK ⁶⁾ 0.548 ± 0.002 18.27 0.349 ± 0.006 11.62 1.57 1.70 70 Gs GK ⁷⁾ 0.513 ± 0.086 17.11 0.326 ± 0.010 10.88 1.57 1.70 70 BRCs GK 0.584 ± 0.017 19.46 0.350 ± 0.012 11.66 1.67 1.80 65 Hs GK 0.759 ± 0.086 25.30 0.347 ± 0.013 11.56 2.19 2.36 40 Aj GK 0.568 ± 0.066 18.94 0.316 ± 0.015 10.53 1.80 1.94 59 KcK 0.793 ± 0.028 26.43 0.391 ± 0.044 13.04 2.03 2.19 48 1) Gs: roasted salt, 2) BRCs: salt removed, 3) Hs: Hanjugeum, 4) Aj: fish sauce 5) KcK: Chinese cabbage kimchi (with salt removed), 6) OGK: organic Green kimchi, 7) green kimchi

Example 5: in vitro Anticancer effect of green kimchi by MTT assay

Step 1: Anticancer Effect of Salted Pickled Green Kimchi by Salt Type

In this embodiment, to find out the anticancer effect according to the type of salt used in salted salted salt, roasted salt, roasted salt, salted water with natural salt, and fermented with green kimchi using general salt without salted water to optimize the optimum period (pH 4.2). The functionality of the kimchi juice was investigated.

Reagents and equipment were RPMI 1640, fetal bovine serum (FBS), 0.05% trypsin-0.02% EDTA and 100 units / ml penicillin-streptomycin for cell culture from GIBCO (USA). Cell culture was performed using a CO ₂ incubator (Forma, model MCO96, Japan). The cell line used was AGS human gastric adenocarcinoma cells obtained from the Korea Cell Line Bank (Seoul National University of Medicine) and cultured. AGS was incubated in a 37 ° C., 5% CO ₂ incubator using RPMI 1640 containing 100 units / ml penicilin-streptomycin and 10% FBS. The cultured cancer cells were refeeded 2-3 times a week, washed with PBS in 6-7 days, and then centrifuged by separating cells attached with 0.05% trypsin-0.02% EDTA. The medium was added to the cancer cells and mixed well so that the cancer cells were evenly distributed by a pipette and used in the experiment while subcultured every 6 to 7 days. Each passage number was recorded during passage and when the passage number was 10 or more times, new cancer cells were re-cultured in a liquid nitrogen tank.

In the MTT assay, cultured cancer cells were seeded in a 96 well plate at 1.0 × 10 ⁴ cells / mL per well, samples were added according to concentrations, and then cultured in a 37 ° C. and 5% CO ₂ incubator. After 72 hours, 20 μl of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazoliumbro mide (MTT) was added and further incubated for 4 hours, and the resulting formazan crystals were dissolved in DMSO. Absorbance was measured at 540 nm.

As a result, as shown in Table 5, the anticancer effect of green kimchi juice by salt type was 99.6% when roasted salt leek kimchi juice was treated with 10.0 μl. Cancer cell survival was inhibited. In the case of baked salt kimchi, the activity increased in concentration-dependent manner to 88.2% when treated with 2.5µl, 94.6% when treated with 5.0µl, and 99.6% when treated with 10.0µl. This increased, and overall showed very high cancer cell survival inhibitory activity of 65-99%.

MTT Analysis of AGS Human Gastric Cancer Cells of Methanol Extracts from Pa Kimchi Prepared with Various Salts Sample OD ₅₄₀ 2.5 µg / assay 5.0 µg / assay 10.0 µg / assay Control 1.267 ± 0.02 ^a 1.267 ± 0.02 ^a 1.267 ± 0.02 ^a Baked salt GK ¹⁾ 0.150 ± 0.01 ^g (88) ³⁾ 0.068 ± 0.01 ^h (95) 0.005 ± 0.01 ^l (99) Roasted Salt GK 0.374 ± 0.01 ^c (71) 0.245 ± 0.01 ^e (81) 0.092 ± 0.01 ^h (93) BR ²⁾ -Natural Salt GK 0.437 ± 0.01 ^b (66) 0.269 ± 0.01 ^d (79) 0.175 ± 0.01 ^f (86) Natural Sea Salt GK 0.446 ± 0.01 ^b (65) 0.266 ± 0.01 ^d (79) 0.138 ± 0.01 ^g (89) 1) GK: Green onion kimchi, 2) BR: Sea water removal, 3) a to i: Different characters represent significant differences by Duncan's multiple range test (p <0.01)

Second Process: Anticancer Effect of Green Kimchi Prepared by Pickling and Salting

In this process, green kimchi prepared by pickling method, roasted salt, natural salt of salted water removed, green kimchi prepared by salting method using Hanjugeum, green kimchi prepared using roasted salt and organic leek, and standardized Chinese cabbage kimchi recipe [Joe Eunju: Chinese cabbage kimchi] Standardization and cancer prevention effect. Methanol extract was prepared by lyophilizing kimchi (pH 4.2) of ripening season using general cabbage kimchi using Pusan National University Ph.D. dissertation (1999)] and in vitro anticancer activity was investigated using AGS human gastric cancer cells. The reagents and equipment used were the same as in the first step, and the MTT assay method was performed in the same manner.

As a result, as shown in Table 6, the anticancer effect of methanol extract of green onion kimchi increased anticancer activity in all samples, and the high concentration treatment group of roasted salt organic onion kimchi, roasted salt general onion kimchi, and liver dehydrated natural salt kimchi (50㎍) / assay) showed a high activity of 85%, 84%, 83% respectively. In case of sauerkraut kimchi, the inhibition rate was lower than that of saline kimchi, but the inhibition rate was higher than that of Chinese cabbage kimchi.

MTT Analysis of AGS Human Gastric Cancer Cells from Methanol Extract from Pa Kimchi Sample OD ₅₄₀ 25 µg / assay 50 µg / assay Control 0.705 ± 0.01 ^a 0.705 ± 0.01 ^a Baked salt OGK ¹⁾ 0.233 ± 0.02 ^de (88) ⁵⁾ 0.107 ± 0.03 ^f (85) Baked salt GK ²⁾ 0.229 ± 0.01 ^de (68) 0.108 ± 0.04 ^f (81) BR ³⁾ -Natural Salt GK 0.238 ± 0.01 ^cde (66) 0.119 ± 0.01 ^f (83) Korean Address GK 0.270 ± 0.04 ^bcd (62) 0.185 ± 0.01 ^e (74) Anchovy fish sauce GK 0.293 ± 0.04 ^bc (58) 0.208 ± 0.04 ^e (71) KC Kimchi ⁴⁾ 0.318 ± 0.03 ^b (55) 0.275 ± 0.04 ^bcd (61) 1) OGK: Organic Green Kimchi, 2) GK: Green Kimchi, 3) BR: Remove Water, 4) KC: Chinese Cabbage 5) a to f: Different characters represent significant differences by Duncan's multiple range test (p <0.01)

Example 6: Peripheral Blood Cells Using Reticulocytes in vivo Micronucleus experiment

In this example, baked salt, Korean kimchi, hot water-picked green kimchi, anchovy pickled green kimchi, and general cabbage kimchi for each sample group, 1,000 mg / kg and 500 mg / kg in two concentrations, once orally administered to mice for 6 hours. After 48 hours infusion of micronucleus-inducing mitomycin C (MMC) into micronucleus-induced mice, blood was collected from the tail vein of the mouse, and the incidence of micronucleus was counted to determine the anticancer effect of pakimchi. Was investigated. Mice were male and male 6-week-old ICR mice (Korea Chemistry Research Institute, Daejeon). Experimental animals weighing about 35 g were used, and feed was raised as standard feed. During breeding, water and feed were supplied in sufficient quantities, and the animal laboratory maintained a temperature of 22 ± 1 ° C and a humidity of 55 ± 5% and maintained a light-dark cycle at 12 hour intervals. A positive control mitomycin C (MMC, 0.1 mg / mL) was purchased from the US Sigma company and dissolved in physiological saline to intraperitoneal injection to 0.1mL / 10g body weight. Samples were prepared using sterile distilled water and 500 or 1000 mg / kg mice were administered orally. The acridine orange-coated slide is an acridine orange solution 10 dissolved in distilled water at a concentration of 1 mg / mL in the center of a pre-heated slide glass at 70 ° C. After dropping the μL uniformly coated with a glass rod, dried and stored at room temperature until sealed and used. Samples were taken 5 μL of blood from the tail blood vessels of the mouse 48 hours after MMC administration, dropped on an acridine oreange-coated slide, covered with cover glass, and then at 4 ° C. for 2 hours. It was prepared by allowing the cells to sufficiently react with acridine orange. The slide was observed at 40 × 10 times by fluorescence microscope (Olympus, model U-ULH, Japan). The reticulocytes counted 2000 from type I to type III, among which reticulocytes with micronuclei were counted. Micronuclear incidence was determined.

As a result, as shown in Figure 9 showed a high incidence of micronucleus in the control group administered only MMC, but when the green kimchi samples orally administered 1000mg / kg and 500 mg / kg, respectively, concentration-dependent suppression of micronucleus induction of MMC The baked salted kimchi 1000mg / kg administration group showed significant antinucleus-inducing ability. The inhibitory effect was higher in the grilled salt leek kimchi group than in the traditional pickled leek kimchi group, and lower in the Hanju Geumpa Kimchi group. Compared with Chinese cabbage kimchi, it was found that roasted salt leek kimchi had high anti-nucleated effect, and that kimchi had higher cancer prevention effect than Chinese cabbage kimchi even in vivo .

Example 7: Investigation of anticancer effect of green kimchi by solid cancer growth inhibition

In the present embodiment, to confirm the direct cytotoxicity of each of the sauerkraut kimchi samples prepared with roasted salt, sun-dried sea salt, salted green kimchi, sauerkraut-pickled green kimchi, and salt-free sea salted kimchi. After performing the cytotoxicity test (viability test) in vitro using the dye exclusion method in order to see the anti-tumor effect of the green kimchi sample, after transplanting the tumor cells in the subcutaneous subcutaneous of the left mouse of the mouse for 20 days Thirty-two days after administration, the mice were dissected to isolate tumors and weighed.

Animals used in this example were male Balb / c mice (Korea Chemistry Research Institute, Daejeon), and weighed about 25 g. Feed was raised as standard feed. During breeding, water and feed were supplied in sufficient quantities, and the animal laboratory maintained a temperature of 22 ± 1 ° C. and a humidity of 55 ± 5% and maintained a light-dark cycle every 12 hours.

Sarcoma-180 tumor cells were passaged and preserved at weekly intervals in the abdominal cavity of Balb / c mice using sarcoma-180 cells as experimental tumor cells. That is, sarcoma-180 cells cultured for one week in the abdominal cavity of experimental animals were taken with ascites and centrifuged (1,200 rpm, 10 min.) With PBS (phosphate buffered saline) to separate tumor cells. The separated cells were suspended in PBS again, centrifuged again to remove the supernatant, and tumor suspensions were made to 1.0 × 10 ⁶ cells / mL. The kimchi samples were prepared using sterile PBS, and the dosage was 1 mg / kg mouse, and the control group was administered only sterile PBS.

Hereinafter, the cytotoxicity test and MTTassay will be described in detail by dividing by process.

Process 1: Cytotoxicity Test

Viability test of this process was performed using 1 mL (2.0 × 10 ⁵ cells) of tumor cell suspension prepared in a disposable 24well plate and RPMI medium of 20% HFCS (heat inactivated fetal calf serum: Gibco Lab., USA). 1 mL was added. 100 μl of sample was added thereto and incubated in 37 ° C. and 5% CO ₂ incubator for 24 hours. After incubation, 50 µl of the cells were mixed well with 50 µl of 0.2% trypan blue solution, and the total number of cells and the number of non-viable and non-stained cells using a hemocytometer. Was measured, and then the cytotoxicity ratio was calculated by comparison with the control cell group without the sample. The formula is as follows.

As a result, as shown in Table 7, the survival rate decreased at concentrations of 1.0 mg / mL and later. Therefore, the following solid cancer growth inhibitory effect experiments were carried out within a range that did not significantly affect sarcoma-180 tumor cells by selecting the concentration.

Cytotoxicity Test of Sarcoma-180 Cells in Culture Medium Containing Methanol Extracted from Pa Kimchi Sample Final concentration (mg / mL) Total cell count (× 10 ⁴ ) Cell viability (%) Control 15.0 ± 3.4 ²⁾ 96.0 Grilled salted green onion kimchi 0.1 12.4 ± 2.3 95.2 0.25 12.4 ± 1.5 90.3 0.5 20.8 ± 5.7 88.4 0.75 15.4 ± 2.8 88.1 1.0 13.8 ± 4.0 86.9 2.0 14.8 ± 1.8 71.2 BR ¹⁾ -Cheonil Salted Kimchi 0.1 14.4 ± 1.9 95.8 0.25 11.2 ± 2.3 94.6 0.5 11.4 ± 2.9 89.4 0.75 10.6 ± 1.6 88.6 1.0 12.6 ± 2.9 85.7 2.0 16.4 ± 5.1 74.3 Hanju Geumpa Kimchi 0.1 16.6 ± 7.2 92.7 0.25 16.2 ± 2.2 92.5 0.5 14.4 ± 3.4 87.5 0.75 12.2 ± 2.7 90.1 1.0 16.4 ± 2.4 85.3 2.0 16.8 ± 4.0 75.0 Anchovy sake leek kimchi 0.1 13.4 ± 3.9 91.0 0.25 12.6 ± 1.9 90.4 0.5 12.2 ± 1.9 90.1 0.75 14.0 ± 3.3 87.1 1.0 12.4 ± 1.7 84.8 2.0 13.6 ± 2.4 73.5 BR ¹⁾ -Cheonil salted cabbage kimchi 0.1 13.2 ± 3.4 90.9 0.25 11.8 ± 1.9 89.8 0.5 10.8 ± 0.7 88.9 0.75 18.4 ± 4.3 86.4 1.0 12.2 ± 2.5 85.2 2.0 16.2 ± 4.1 74.0 [Note] 1) BR: Remove Beast 2) Repeat 3 times ± SD

Second Process: Weighing Solid Rock

Six animals per group were used to subcutaneously implant 0.2 mL (6.0 × 10 ⁶ cells / mouse) of tumor cell fluid stored in the laboratory at weekly intervals into the left groin for 24 hours. Sample solutions were injected intraperitoneally once daily. On the 26th day of tumor cell transplantation, the solid tumors generated by lethality were extracted and weighed, and then the tumor growth inhibition ratio (IR:%) was calculated according to the following equation.

Cw: mean tumor weight of control group

Tw: mean tumor weight of treatment group

As a result, as shown in Table 8, the control group transplanted with only tumor cells had a tumor weight of 7.37 g, whereas the weight of the group treated with roasted sake lees decreased to 5.37 g, showing a 27.5% tumor suppression effect. In addition, the group treated with sesame removed kimchi and fish sauce showed 18% tumor suppression effect. However, the administration of Hanju Geumpa kimchi and general cabbage kimchi showed no significant difference from the control group transplanted with tumor cells only (p <0.05).

Weighing Results of Solid Carcinoma in Balb / c Mice with Sarcoma-180 Cells Sample Tumor weight (g) Inhibition rate S-180 + PBS 7.37 ± 0.55a4) - S-180 + grilled salt GK1) 5.34 ± 0.94c 27.5 S-180 + BR2) -Sun Salt GK 5.98 ± 0.90bc 18.9 S-180 One Address GK 6.90 ± 1.43ab 6.4 S-180 + Anchovy Sauce GK 6.03 ± 0.70bc 18.2 S-180 + Kc Kimchi 3) 6.95 ± 0.25ab 5.7 [Note] 1) GK: Pa Kimchi, 2) BR: Ganjo, 3) Kc Kimchi: Chinese cabbage Kimchi 4) Value of 3 times ± SD a ~ c: Other characters indicate significant differences by Duncan's multiple range test. (p <0.05)

Third Process: Change in Weight of Organs

In this process, PBS was injected into the tumor cell group to measure the action in other organs of the kimchi samples, and the sample treatment group was injected by injection with the same amount of methanol extract for 20 days after tumor cell transplantation. The change in weight was measured.

As a result, as shown in Table 9, the weight ratio of liver which is responsible for detoxification of various harmful substances generated during metabolism is different from the control group injected with PBS only, the group treated with Han Geumpa Kimchi, and the group injected with Chinese cabbage kimchi after treatment with cancer cells. Compared with the group, the spleen also showed the same trend. Organs such as heart and kidney did not differ significantly between groups.

The weight of each organ of Balb / c mice fed with kimchi samples Sample Weight (g) Spleen / body (%) Liver / Body (%) Heart / body (%) Height / Body (%) Control 23.9 ± 1.30 0.46 ± 0.19 5.30 ± 0.97 0.42 ± 0.02 1.47 ± 0.25 S180 + PBS 26.5 ± 2.55 0.78 ± 0.32 5.85 ± 0.99 0.38 ± 0.04 1.09 ± 0.44 S180 + Baked Salt OGK ¹⁾ 26.1 ± 2.14 0.69 ± 0.14 5.77 ± 0.93 0.39 ± 0.03 1.28 ± 0.28 S180 + BR ²⁾ -Sun Salt GK 26.0 ± 1.81 0.63 ± 0.29 5.25 ± 0.36 0.39 ± 0.03 0.98 ± 0.39 S180 + One Address GK ³⁾ 27.7 ± 2.76 0.70 ± 0.28 6.61 ± 1.18 0.36 ± 0.04 1.15 ± 0.14 S180 + Anchovy Sauce GK 25.5 ± 1.47 0.69 ± 0.35 6.18 ± 1.15 0.46 ± 0.19 1.37 ± 0.27 S180 + KcK ⁴⁾ 27.5 ± 1.07 0.80 ± 0.14 7.11 ± 1.09 0.36 ± 0.01 1.24 ± 0.24 1) OGK: Organic Green Kimchi, 2) Seaweed Removal, 3) GK: Green Kimchi, 4) KcK: Chinese Cabbage Kimchi

Example 8: Enhancement of NK cell immune activity of green kimchi

The same mouse used for investigating the anticancer effect of green kimchi by solid cancer growth inhibition in Example 7 was used to measure the activity of spontaneous killer (NK) cells in the spleen to investigate the immune activity enhancement effect of green kimchi.

The spleens of Balb / c mice were removed aseptically in a clean bench with 5 mL of RPMI 1640 medium containing penicillin (100 U / mL) and streptomycin (100 μg / mL). After washing three times, finely ground. The cell suspension was filtered through a 70 μm nylon mesh, centrifuged to collect lymphocytes, suspended in media, and then centrifuged using histopaque-1077 (500 × g, 30 min, 18 ° C.) to separate lymphocytes. Effector cells were resuspended in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamine, penicillin (100 U / mL), streptomycin (100 μg / mL), This was incubated for 1 to 2 hours at 37 ° C. in a CO ₂ incubator to allow cells to adhere to the culture flask (60 mm dish). Non-adherent natural killer (NK) cells were collected by centrifugation, resuspended in culture medium, and the appropriate number of cells was counted.

The activity of natural killer (NK) cells was measured using the MTT method of Example 5 mentioned above. Yac-1 mouse lymphoma cells were used as target cells. Inoculate effector cells at 200 × 10 ⁴ cells / mL and 100 × 10 ⁴ cells / mL with Yac-1 cells 5 × 10 ⁴ cells / mL as target cells (the ratio of effector cells to target cells is 40: 1 and 20 : 1). That is, Yac-1 cells were prepared in 5 × 10 ⁴ cells in RPMI 1640 medium containing 10% FCS, 2 mM L-glutamin, penicillin [penicillin (100 U / mL)], and streptomycin (100 μg / mL). After diluting to a density of / mL, 50 μl was added to each well. NK cells were suspended at a density of 1 × 10 ⁷ cells / mL and 50 μl each was added to 96well flate-bottomed microtire plates. After it was for 3 days in 37 ℃, CO ₂ incubator 10㎕ by the MTT (5㎎ / mL) solution was added to each well and then were incubated at 37 ℃ 4 hours. 25 μl of SDS (10% in 0.02N HCl) was added thereto, followed by color development at room temperature for 30 minutes, and then OD was measured at 540 nm. Cell cytolysis percentage was calculated by the following formula.

As a result, as shown in FIG. 10, NK cell activity was low in the PBS-only group after cancer cell treatment, but the activity was increased in the group to which the sample was administered. In particular, the roasted salt leek kimchi injection group showed high activity of more than 80% at the ratio of NK cell and Yac-1 cell at 40: 1. Next, the liver-derived natural salt kimchi injection group and the salted-pickled kimchi injection group showed high activity, and compared with the cabbage kimchi using the same salt, the kimchi showed higher NK cell activity. Even in the case of cancer cell transplantation, the NK cell activity is increased in this way, and thus, it has an anticancer effect in terms of human immunity.

Example 9 Changes of Enzyme Activity in the Liver

Hepatic glutathione S-transferase activity was measured in the same mouse which investigated the anticancer effect of green kimchi by solid cancer growth inhibition in Example 7, and the detoxifying effect of green kimchi was investigated.

After the mice were killed, the liver was perfused with saline of 4 ° C. or less to remove blood remaining in the liver, and the liver was extracted. 0.1 M potassium phosphate buffer (pH 7.4) was added per gram of liver tissue and ground with a glass teflon homogenizer under ice cooling. The homogenate fraction was used as a homogenate fraction, which was centrifuged at 13,000 rpm for 10 minutes to remove the nucleus and unbroken cell sections, and the supernatant obtained by ultracentrifugation at 105,000 × g for 1 hour was used as a psychosol ( cytosol) fraction. The homogenate fraction thus obtained was used to measure glutathione content, and the cytosol fraction was used to measure glutathione-S-transferase activity.

Hereinafter, a specific method is demonstrated according to the following process.

First Process: Glutathione S-Transferase Activity Change

75 μl of 0.04 M reduced glutathione and 0.1 mL of enzyme solution were added to 0.1 M potassium phosphate buffer (pH 6.5), and 0.5 mL of 20% trichloroacetic acid was added to the blank. The enzyme was inactivated and then reacted at 25 ° C. for 5 minutes. 25 μl of 0.12M 1-chloro-2,4-dinitrobenzene was added to the blank and the sample, followed by reaction at 25 ° C. for 2 minutes, and 20% trichloroacetic acid was added to the sample to complete the reaction. The absorbance was measured at 340 nm, and the activity was calculated using the mole absorption coefficient of 9.6 mM ^-1 cm ^-1 of 1-chloro-2,4-dinitrobenzene. . The unit of enzyme activity was expressed as nmole number of 2,4-di-nitrobenzene-glutathione produced by mg protein for 1 minute.

As shown in FIG. 11, glutathione-S-transferase activity was 433.3 nmol / mg protein / min in the control group without tumor cells and dropped to 264.9 nmol / mg protein / min in the control group in which tumor cells were transplanted. lost. In the case of the sample injection, the activity was in the order of baked salt leek kimchi group 397.4, dehydrated natural salted kimchi group 318.7, pickled salted kimchi group 302.2, general cabbage kimchi group 280.3, Hanju Geum kimchi group 273.0nmol / mg protein / min Glutathione-S-transferase activity was found to be significantly higher in the roasted salt leek kimchi group, and when the same salt was used, the kimchi showed higher activity than the cabbage kimchi.

Second Process: Glutathione Content Change

4% sulfosalicylic acid was added to the enzyme source (400-600 µg protein) as a protein reagent and 5,5 in disulfide reagent [0.1M sodium phosphate buffer (pH8.0)] Dissolve 2-dithiobis (2-nitrobenzoic acid)] 2.7 mL of blue color was measured at 412 nm and calculated by standard curve. The unit was expressed as glutathione μmole per gram of tissue.

As a result, as shown in FIG. 12, the glutathione content of the normal control group was 0.366, and the S-180 + PBS group was 0.232, while the grilled salt leek kimchi group showed a high glutathione content of 0.488, followed by 0.376 salted kimchi with salted water Green onion kimchi 0.315, Korean cabbage kimchi 0.227, Hanju geumpa kimchi 0.265. The salted pickled kimchi had higher glutathione content than the salted pickled kimchi, except for the kimchi made with Hanjugeum. The green kimchi with the same salt showed the higher glutathione content.

As described above, the present invention kimchi marinated in saline instead of salted anchovy as described through the above embodiment has excellent taste as well as antimutagenic and anticancer effects. The kimchi of the present invention is excellent in preservation when it is packaged in a flexible aluminum wrapping paper with a gas absorbent and stored at 10 ° C. Industrialization is advantageous, and this is a very useful invention in the functional pakimchi food industry.

Claims (4)

Marinate the chives in 5% brine for 30 minutes and shake off the water, and then add salted anchovy fish sauce 10.0, red pepper powder 9.7, garlic 5.2, ginger 2.7, sesame seeds 1.3, sugar 1.5, and glutinous rice 12.0 in a relative ratio to 100.0. In the method for producing anti-cancer functional green kimchi, which is mixed with chopped chives and then fermented by wrapping 5-8 strands of green onions or packaging as it is, wherein the saline is made of roasted salt. delete An anticancer functional green kimchi prepared by the method of claim 1. A method of preserving anti-cancer functional kimchi, characterized in that the kimchi according to claim 3 is packaged in an aluminum wrapping paper with a gas absorbent Ca (OH) ₂ , and stored at 10 ° C. for distribution.