KR101100723B1 - Kimchi using black garlic and phelliunus linteus - Google Patents

Abstract

The present invention relates to kimchi for enhancing biological activity using black garlic and situation mycelia powder.

Kimchi for physiological activity-enhancing Bossam according to the present invention 13-17% by weight, red pepper powder 4.5-5.5% by weight, white sugar 2.4-2.5% by weight, black garlic 1.7-1.9% by weight, brown rice brown mycelium powder 1.8-2.2% by weight, 1.4-1.6% by weight of pears, 1.4-1.6% by weight of buttercups, 1.4-1.6% by weight of salted shrimp, 1.4-1.6% by weight of onions, 0.9-1.1% by weight of ginger, 0.6-0.8% by weight of green onions, 0.4-0.6% of sodium L-glutamate By weight, sesame oil 0.25 ~ 0.35% by weight, pine nut 0.08 ~ 0.12% by weight, sesame 0.08 ~ 0.12% by weight, 5'- ribonucleotide disodium 0.015 ~ 0.025% by weight and the remaining amount of pickled cabbage.

Kimchi, black garlic, situational mycelium

Description

Kimchi for Enhancing Biological Activity Using Black Garlic and Situational Mycelia {KIMCHI USING BLACK GARLIC AND PHELLIUNUS LINTEUS}

The present invention relates to kimchi, and more particularly, to kimchi for supplementary kimchi with enhanced physiological activities such as antioxidant, immune and anticancer activity by adding black garlic and situation mycelia powder to existing kimchi.

With the rapid development of science and technology and the development of related industries, our daily lives have changed greatly in terms of comfort and convenience, especially the changes in diet. Despite these changes, Kimchi, one of our own foods, is still a representative traditional fermented food that cannot be missed from our table.

Kimchi is a shelf-life food with various flavors and flavors, mixed with various kinds of pickled vegetables to prevent rancidity and deterioration, and the effects of lactic acid bacteria and fermentation bacteria. The sour, irritating carbonated flavors blend well with a unique flavor.

As a source of dietary fiber, vitamins, minerals, etc. in Kimchi, the antimutagenic activity of Kimchi's dietary fiber and garlic, ginger, and red pepper, which are ingredients of Kimchi, is known to prevent colon cancer, anemia, and arteriosclerosis. The results of studies on important physiological and physiological effects have been reported.

On the other hand, the general efficacy of garlic (hereinafter referred to as white garlic) is known to be effective in preventing and restoring cardiovascular diseases such as stroke, hypertension, and arteriosclerosis, rejuvenation, promoting blood circulation, neurostable action, parasite control and bactericidal effect. have. Recently, studies have shown that sulfur-containing amino acids and derivatives thereof are involved in the detoxification of toxic and heavy metal poisoning in vivo and inhibit heavy metal absorption in the digestive tract, and are effective in improving antimicrobial activity and lipid metabolism in the body. Not only is this experimentally proven, it has been selected by the US National Cancer Institute as the most effective food ingredient for cancer prevention.

Vitamin B1, also known as anti-fatigue vitamin, acts as white garlic to form a substance called alliamine, also called garlic vitamin. This vitamin is known to be effective for maintaining metabolism, maintaining fatigue and improving stamina, such as vitamin B1.

  Despite the various physiological functions of white garlic, allin, the main component of white garlic, breaks down into allicin by allinase when the tissue of the white garlic breaks down, causing a unique smell. Causes bad breath.

For the same reason, while knowing the various excellent effects of white garlic is often avoided edible, it is known that excessive intake of white garlic may be rather harmful to health by stimulating the stomach and liver.

There are several ways to remove the spicy taste of white garlic, and typically, a method of removing odor by inactivating the enzyme kinase by heat treatment of the unique odor component of white garlic may be proposed. However, when the heat treatment is removed to remove the spicy taste of white garlic, active ingredients such as vitamin A, vitamin C, potassium, phosphorus, sulfur, selenium and various amino acids possessed by white garlic are destroyed.

A product that is fermented to remove the spicy taste of white garlic without losing the physiological activity of white garlic is called black garlic, which is obtained by self-fermentation at a certain temperature and humidity without using any additives. The unpleasant odor peculiar to white garlic is completely disappeared, and the antioxidant power is known to increase by about 10 times more than the raw garlic without destroying the original active ingredient of white garlic.

In addition, black garlic produces S-allyl cysteine, which is effective in preventing cancer, inhibiting cholesterol, improving arteriosclerosis, preventing heart disease, and preventing Alzheimer's disease, and inhibiting cholesterol oxidation, inhibiting free radicals, and preventing arteriosclerosis. It is known that the amount of polyphenol compounds present is greatly increased.

On the other hand, mushrooms are valuable biological resources that exist in the tens of thousands of species on earth. Since ancient times, they are known to be effective as tonic, soothing, lowering blood pressure, strong heart, diuresis, hepatitis as well as incurable diseases such as cancer. . Recently, it has been found that fruit fruit extracts and mycelium cultures of mushrooms are effective in improving the constitution and preventing and treating various diseases, and the use of them as medicines and health foods is increasing.

In particular, mushrooms have various effects such as antibacterial action, antiviral action, cholesterol lowering action, hypoglycemic action, hypotension action, antithrombotic action, interferon induction action, immunopotentiation action and antitumor action. Industrial interest in exploration, development and use is very high.

  Currently, schizophyllan, lentinan, PS-K (or krestin) are extracts from mycelium and are already available as immune boosters. (Hericium erinaceum), Cordyceps sinensis, and other mushrooms have active biological research because they have functional food materials or high added natural physiological activity.

An object of the present invention is to replace the white garlic included in the kimchi with black garlic, and to add the sapling mycelium powder, to provide a supplementary kimchi for enhanced physiological activities such as antioxidant, immune and anticancer activity.

Kimchi for enhanced physiological activity using black garlic and situation mycelium according to the present invention for achieving the above object as kimchi added black garlic and situation mycelium powder, 1.7 ~ 1.9% by weight of black garlic and 1.8 ~ 2.2% by weight of brown rice mycelium powder Characterized in that it comprises a.

 Preferably, physiological activity-enhancing kimchi using black garlic and situation mycelium according to the present invention 13-17% by weight of pickled radish, red pepper powder 4.5-5.5% by weight, 2.4-2.5% by weight of sugar, 1.7-1.9% by weight of black garlic, Brown Rice Strains Mycelium Powder 1.8-2.2%, Pear 1.4-1.6%, Buttercup 1.4-1.6%, Salted Shrimp 1.4-1.6%, Onion 1.4-1.6%, Ginger 0.9-1.1%, Chives 0.6-0.8 Wt.%, L-Glutamate 0.4 to 0.6%, Sesame oil 0.25 to 0.35%, Pine nut 0.08 to 0.12%, Sesame 0.08 to 0.12%, 5'-ribonucleotide disodium 0.015 to 0.025% and remaining pickled cabbage It includes.

Physiological activity-enhancing Bossam kimchi according to the present invention has the advantage to improve the antioxidant, immune and anticancer activity compared to the existing kimchi by adding black garlic and situation mycelia powder to the existing kimchi.

Advantages and features of the present invention and methods for achieving them will be apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various different forms, and only the embodiments make the disclosure of the present invention complete, and those skilled in the art to which the present invention pertains. It is provided to fully inform the person having the scope of the invention, which is defined only by the scope of the claims.

Hereinafter will be described in detail with respect to physiological activity-enhancing Bossim kimchi using black garlic and situation mycelium according to a preferred embodiment of the present invention.

Kimchi for physiological activity-enhancing Bossam according to the present invention is a pickled cabbage as the main material, black garlic and situation mycelium powder is added to seasoning cattle.

Black garlic is immune active while maintaining many benefits such as prevention of cardiovascular diseases such as stroke, hypertension and arteriosclerosis, recovery of vitality, promotion of blood circulation, neurostable action, parasite control and bactericidal effects of garlic (hereinafter referred to as raw garlic). It appears similar or higher than raw garlic, and has the advantage of eliminating the smell of garlic.

Situation mycelia powder increases the antioxidant activity in kimchi, improves the ability to inhibit lipid peroxidation, and also plays a role in supplementing the kimchi sensory caused by black garlic. The situation mycelium powder may be brown rice situation mycelium powder powdered by inoculating mushroom mushrooms in brown rice, but is not limited thereto, and any one sold in the name of situation mycelium powder, mushroom mushroom mycelium powder, etc. may be used. .

In the kimchi according to the present invention, the black garlic is included in the content ratio of 1.82% by weight of the total weight of kimchi, the situation mycelium powder is most preferably included in the content ratio of 2% by weight, when considering the error range, black garlic 1.7 ~ 1.9. The content of the weight% and 1.8 ~ 2.2% by weight mycelial powder can be presented.

As a result, when black garlic and situation mycelia powder was added to kimchi in the above content ratio, it was confirmed that satisfactory workability and sensory activity, and physiological activities such as immunity and anticancer activity, particularly antioxidant such as lipid peroxidation inhibitory activity, were the best. On the other hand, when black garlic and situation mycelium powder is added excessively, there is a problem that the functionality of kimchi is lowered even though the physiological activity is increased.

Optimized examples of Kimchi for physiological activity-enhancing Bossam according to the present invention, pickled cabbage 65%, pickled radish 15% by weight, red pepper powder 5% by weight, white sugar 2.46% by weight, black garlic 1.82% by weight, brown rice flour mycelium powder 2%, 1.5% of pears, 1.5% of buttercups, 1.5% of salted shrimps, 1.5% of onions, 1% of ginger, 0.7% of chives, 0.5% of sodium L-glutamate, 0.3% of sesame oil, 0.1% of pine nuts, sesame seeds 0.1% by weight, 0.02% by weight of 5'-ribonucleotide disodium can be given.

However, 5 ~ 10% error may occur when adding each material. In consideration of this, kimchi for enhancing biological activity according to the present invention kimchi pickled radish 13-17% by weight, red pepper powder 4.5-5.5% by weight, white sugar 2.4-2.5% by weight, black garlic 1.7-1.9% by weight, brown rice microorganism powder 1.8 ~ 2.2 wt%, Pear 1.4-1.6 wt%, Buttercup 1.4-1.6 wt%, Salted shrimp 1.4-1.6 wt%, Onion 1.4-1.6 wt%, Ginger 0.9-1.1 wt%, Scallion 0.6-0.8 wt%, L-glutamic acid Sodium 0.4-0.6%, sesame oil 0.25-0.35%, pine nuts 0.08-0.12%, sesame 0.08-0.12%, and 5'-ribonucleotide disodium 0.015-0.025%, and pickled cabbage is included do.

As a result of comparing the bioactive kimchi prepared according to the present invention and the conventional kimchi containing black garlic and brown rice mycelium powder instead of raw garlic instead of the present invention, in the case of the bioactive kimchi according to the present invention, workability and functionality It was excellent in physiological activity characteristics such as antioxidant, immunity, anticancer activity without being lowered compared to the prior art.

Example

Hereinafter, the action and effect of the physiological activity-enhancing Bossim kimchi according to the present invention through the embodiment will be described in more detail.

1. Manufacture of Kimchi

Experimental group

Pickled radish 15% by weight, red pepper powder 5% by weight, white sugar 2.46% by weight, black garlic 1.82% by weight, brown rice flour mycelium powder 2% by weight, pear 1.5% by weight, buttercup 1.5% by weight, salted shrimp 1.5% by weight, onion 1.5% by weight, Prepare seasoning beef by mixing 1% by weight of ginger, 0.7% by weight of chives, 0.5% by weight of sodium L-glutamate, 0.3% by weight of sesame oil, 0.1% by weight of pine nuts, 0.1% by weight of sesame and 0.02% by weight of 5'-ribonucleotide disodium. To prepare kimchi, tossed with seasoning beef prepared in pickled cabbage 65% by weight.

Table 1 shows the compounding ratio of the physiologically active reinforced kimchi.

TABLE 1
Raw material name Compounding ratio (% by weight) Raw material name Compounding ratio (% by weight) Pickled cabbage 65.00 onion 1.50 Pickled vegetables 15.00 ginger 1.00 chili powder 5.00 Sect 0.70 White sugar 2.46 Sodium L-Glutamate 0.50 Brown Rice Situation 2.00 Sesame oil 0.30 Black garlic 1.82 Pine nuts 0.10 ship 1.50 Sesame 0.10 Buttercup 1.50 5'-ribonucleotide disodium 0.02 Salted shrimp 1.50 Sum 100.00

Comparison

Kimchi was prepared in the same blending ratio as that of the experimental group, except that 7.28 wt% of fresh garlic was added without adding black garlic and brown rice mycelium powder, and the blending ratio of pickled cabbage was 61.54 wt%.

end. Total bacteria

Changes in Total Bacterial Count According to Storage Period of Experimental Kimchi Prepared by Addition of Black Garlic to 1.82% by Weight to 25% of Raw Garlic and 2% by Weight of Strain Mushroom Mycelium Powder and Controlled Kimchi without Raw Garlic The measurement results are shown in Table 2. Hereinafter, in the tables and drawings, the control means a control to which black garlic and situational mycelium powder are not added.

[Table 2] (unit: CFU / g)

Referring to Table 2, the total bacterial count immediately after preparation of the control group was 1.2 × 10 ⁵ CFU / g, gradually increased at the beginning of storage, and increased with longer storage period, and prepared at 2.1 × 10 ⁸ CFU / g at 15 days of storage. It increased about 3 log cycles more than immediately after. The experimental group was 1.7 × 10 ⁵ CFU / g immediately after preparation and appeared to be almost the same number of bacteria compared to the control group.

Changes in storage period showed almost the same bacterial count as the control group except for 7 days of storage.

I. Lactic acid bacteria count

The results of measuring the change in the number of lactic acid bacteria according to the storage period of kimchi prepared by adding black garlic and mushrooms are shown in Table 3.

Table 3 (unit: CFU / g)

Referring to Table 3, the control group without adding black garlic and mushroom mycelium was 1.1 × 10 ⁵ CFU / g immediately after preparation, and 1.5 × 10 ⁵ CFU / g in kimchi prepared with optimum concentration of black garlic and mushroom mushroom mycelium. As the storage period increased, the number of lactic acid bacteria increased with increasing storage period, and showed 1.0 ~ 1.2 × 10 ⁸ CFU / g of bacteria at 15 days of storage.

All. Yeast and Mold Water

The results of measuring the change of yeast and mold number according to the storage period of the experimental and control kimchi are shown in Table 4.

[Table 4] (Unit: CFU / g)

Referring to Table 4, in the case of the control group, the number of yeasts and molds immediately after preparation was 4.9 × 10 ³ CFU / g, and the number of bacteria immediately after preparation of the experimental group was 5.8 × 10 ³ CFU / g. The difference does not appear to be large, as it is an error in sampling.

The change of yeast and mold number according to the storage period decreased until 7 days of storage and then gradually increased again. The storage period of the experimental group prepared with the black garlic and the situation mushroom mycelium showed the lowest bacterial count on the 7th day of storage as in the control group, and did not differ significantly from the control group during the storage period.

As a result, the addition of black garlic and situation mushroom mycelium known as a bioactive substance can be seen that there is little difference in fermentation aging with existing kimchi.

la. Polyphenol Compound Contents according to Storage Period of Kimchi Added with Black Garlic and Situary Mushroom Mycelium

The results of measuring the change in the content of the polyphenolic compound according to the storage period of the experimental group and the control group are shown in Table 5.

In order to measure the content of polyphenol compound, which is one of the compounds exhibiting antioxidant activity, 0.5 mL of Follin-Denis reagent and 1 mL of saturated sodium carbonate were added to 1 mL of the extract (hereinafter referred to as extract) in which the lyophilized extract of each of the experimental and comparative groups was diluted in water. After mixing the (Na 2 CO 3) solution and 7.5 mL of distilled water, the absorbance was measured at 760 nm after 30 minutes, and gallic acid (Sigma, USA) was used as a standard material.

Referring to Table 5, the content of the polyphenol compound in the control kimchi was 115.41 μg / mL immediately after preparation, and the content of the polyphenol compound in the experimental group was 135.22 μg / mL, which was somewhat higher than that of the control group.

According to the storage period, the content of polyphenol compounds in both the control group and the experimental group did not have any particular tendency, and the experimental group contained higher contents than the control group.

[Table 5] (Unit: μg / mL)

hemp. Flavonoid Contents during Storage of Kimchi Added with Black Garlic and Situary Mushroom Mycelium

Table 6 shows the results of measuring the change of flavonoid compound content according to the storage period of the Kimchi in the control group.

In order to determine the flavonoid compound content, one of the compounds exhibiting antioxidant activity, 0.9 mL of 80 vol% EtOH was added to 0.1 mL of the sample extract, and 0.1 mL of 10 vol% aluminum nitrate, 0.1 mL of 1M potassium acetate, and 0.5 mL of this mixture. 4.3 mL of 80 vol% EtOH was added respectively. After leaving the reaction solution at room temperature for 40 minutes, the absorbance value was measured at 415 nm. As a standard material, quercetin (Sigma Chemical Co., USA) was diluted to 0.03%, and the content was calculated by substituting the standard curve.

Referring to Table 6, the flavonoid compounding content of the control group was 11.52 mg% immediately after preparation, and the experimental group kimchi was 14.33 mg%, showing a somewhat higher flavonoid content than the control group.

When the storage period was changed, both the control group and the experimental group tended to decrease gradually as the storage period increased, and it was confirmed that the experimental group maintained a somewhat higher content during the storage period than the control group.

[Table 6] (unit: mg%)

bar. Changes in Chromatographic Values of Kimchi with Optimum Concentrations of Black Garlic and Situary Mushrooms

  The results of measuring chromaticity change according to the storage period of the experimental and control kimchi are shown in Table 7.

Referring to Table 7, the lightness of the control group was 37.42 immediately after preparation and 36.80 in the case of experimental kimchi, slightly lower than the control group. The lightness of the experimental group was relatively lower than that of the control group because the black garlic was added instead of the fresh garlic.

The change in storage period showed a slight increase and decrease during storage, but did not change significantly with the initial storage period. In the case of redness, the control group was 8.84 immediately after manufacture, but the experimental group was slightly lower (8.33). The change according to storage period tended to decrease slightly with increasing storage period.

In the case of yellowness, the control group was 8.90 immediately after manufacture, but in the experimental group, it decreased to redness as 8.05. The change of storage period, like lightness, showed a slight increase and decrease during storage, but did not change significantly with the initial storage period.

TABLE 7

  four. Change of hardness with storage period

The results of measuring the change in hardness according to the storage period of the experimental and control kimchi are shown in Table 8.

[Table 8] (Unit: kg)

Referring to Table 8, the hardness of the control group was found to be 10.09 kg, and even though the storage period increased, the value did not show a tendency to increase or decrease, and there was no difference during the storage period. It seems to be because the thickness is not constant. In the case of the experimental group kimchi, like the control group, there was no specific trend even if the storage period increased, so that there was no difference in texture between the experimental group and the control group.

Ah. Changes in pH and Acidity of Kimchi Added with Black Garlic and Phellinus Mushroom Mycelia

 ① pH

Figure 1 shows the change of pH according to the storage period of the experimental group kimchi added black garlic and situation mushroom mycelium powder.

Referring to FIG. 1, the pH immediately after the preparation of the control group was 5.37, slightly higher than the pH of the experimental kimchi (5.32), but does not appear to be a big difference. The change according to storage period decreased with increasing storage period, which was pH 4.78 at 15 days of storage. The change of pH according to the storage period of the kimchi in the experimental group tends to decrease with increasing storage period.

 ② acidity

Figure 2 shows the change in acidity according to the storage period of the experimental group kimchi added black garlic and situation mushroom mycelium powder.

Referring to FIG. 2, the acidity of the kimchi prepared immediately after the preparation of the control kimchi was 0.46%, and the kimchi prepared by adding the black garlic and mushroom mushroom mycelium was 0.45%, which was not different from the control. The pH of the control group showed that the acidity increased sharply after 4 days of storage, resulting in 0.97% of acidity on 15 days of storage. In the experimental group, the fermentation by adding black garlic and situation mushroom mycelium does not appear to be a problem because it shows similar acidity over the storage period without significant difference from the control group.

character. Change of electron donating ability

Table 9 shows the results of measuring the electron donating ability according to the storage period of the experimental kimchi and the control kimchi.

The electron donating ability of the experimental kimchi and the control kimchi was measured by mixing 2 mL of kimchi extract with 2 mL of 0.2 mM DPPH (1,1-diphenyl-2- picrylhydrazyl), and leaving it for about 30 minutes at room temperature. The absorbance was measured at 517 nm. Electron donating ability (%) was calculated by the following equation.

Electron donating ability (%) = (1- (absorbance of sample addition ÷ absorbance of no addition)) x 100

Referring to Table 9, the electron donating ability was 77.29% immediately after the preparation of the control kimchi, 83.24% in the experimental group was slightly higher than the control group. The change in storage period showed a similar electron donating ability to the control group up to 10 days after storage, and it was 69.96% at 15 days of storage, which tended to decrease slightly from the beginning of storage.

The change of the electron donating ability according to the storage period of the kimchi in the experimental group was slightly higher than that of the control group, and from the 7th day of storage, the electron donating ability was almost similar to the control group.

[Table 9] (unit:%)

car. Changes in lipid peroxidation

The results of measuring the change in lipid peroxidation according to the storage periods of the experimental kimchi and the control kimchi are shown in Table 10.

The degree of lipid peroxidation of mycelium mycelium extract in linoleic acid emulsion matrix was measured by thiocyanate method.

Inhibition of lipid peroxidation (%) = 1-(OD sample ÷ OD control) x 100

Referring to Table 10, in the case of the blank without kimchi, the OD value immediately after the linoleic acid emulsion was prepared was 0.0849, which was lower than that of the experimental groups added with kimchi. However, the storage period increased rapidly after 1 day of storage, and the OD value was 2.7336 very high on the 9th day of storage, indicating that oxidation was very fast.

In the control group, the OD value was 0.1059 immediately after the preparation of the emulsion, which was higher than the blank, but gradually increased as the storage period increased. there was.

On the other hand, in the case of the experimental group, it was 0.1072 immediately after manufacture and it was the highest value in the result immediately after manufacture. From the storage period, it was found that gradually increased than the control group from the 3rd day of storage, and on the 9th day of storage, the OD value was 0.2506, which is about 58% of the control group. The production of lipid peroxide can be suppressed.

Table 10 (Unit: Optical Density at 500 nm)

Ka. Metal ion removal ability

 Table 11 shows the results of measuring the metal ion removal ability according to the storage period of the experimental kimchi and the control kimchi.

Referring to Table 11, the control kimchi was 68.40% of metal ion removal ability immediately after preparation, and the experimental group kimchi was 71.73 mg%, showing a somewhat higher metal ion removal ability than the control. When the storage period was increased, the control group decreased, and it decreased by about 17% to 56.56% on the 15th day of storage. Kimchi in the experimental group showed a slightly higher value during the storage period than the control group during storage, but it does not seem to be a big difference considering the sampling error.

[Table 11] (unit:%)

Get. Comprehensive likelihood

Table 12 shows the results of measuring the overall acceptability of taste, aroma and color according to the storage period of the experimental kimchi and the control kimchi.

Referring to Table 12, the control kimchi immediately after the production of the overall preference of 3.73, the storage period increased as the overall preference increased to 4.18 on the seventh day of storage, after that gradually decreased to a total acceptance of 3.72 on the 15th day of storage.

The reason why the overall acceptability increases and decreases during the storage period is that the kimchi reaches the optimum ripening time around 7 days by fermentation and the acceptability decreases due to acid production. On the other hand, the kimchi of the experimental group showed a slightly lower overall acceptability of 3.58 than the control group immediately after the preparation, and the overall acceptability changed with the trend similar to that of the control group. The reason why the overall acceptability was lower than that of the control group seemed to be due to the non-favorability of the sensory test agents due to the black garlic of black garlic and the yellow color of the situation mushroom mycelium, but there was no significant difference in taste, aroma and texture.

TABLE 12

wave. Immune activity

 a. Mitogen activity

Lymphocyte mitogen activity is measured by MTT assay. In other words, the MTT assay captures the spleen from the ICR mouse, transfers it to a petri dish containing ice cold RPMI-1640 medium, places the metal body (mesh # 100) on the spleen, and releases cells from the spleen while pressing the rubber stopper of the syringe. . The solution was filtered using a metal body (mesh # 200), red blood cells were removed by the stock solution hemolysis method, washed three times with the medium, and the cell concentration was adjusted to 5 × 10 6 cells / mL RPMI-1620, and 90 μL each. Dispense into 96 well plates, add 10 μL of hot-water extracts, and incubate in a 37%, 5% CO ₂ incubator for 3 days. Then, MTT solution at 1 mg / mL concentration was added and reacted for 5 to 6 hours at 37 ° C. in a 5% CO ₂ incubator, and then the supernatant was carefully removed to obtain the resulting MTT formazan precipitate. The obtained formazan was 100 μL of 0.04 Lymphocyte proliferation was measured by absorbance at 570 nm after adding N HCl / isopropanol and dissolving for 5 minutes and adding the same amount of water.

3 is a graph showing the effect of mitogen activity of the hot water extract of the experimental group kimchi and the control kimchi.

Referring to FIG. 3, in the mitogen activity, the hydrothermal extract (Kimchi Control) of the control kimchi showed 1.11 times the activity of the group (Saline Control) treated with saline without a sample, while the hot water prepared from the experimental group Kimchi The extract showed increased activity after 7 days compared to the control kimchi, but did not show a tendency to increase as the fermentation (aging) period was extended. However, the activity of the experimental group kimchi was confirmed to increase than the control kimchi.

As a result, it was confirmed that mitogen activity was increased than conventional kimchi and contributed to immune activity. In addition, the optimum ripening time of the kimchi in the experimental group can be seen that the kimchi after 7 days after the preparation is superior to the kimchi after 15 days.

 b. Myeloid Cell Proliferation through Peyer's Patch

The bone marrow cell proliferation active ingredient was measured by activating the cells obtained from Peyer's patch of the small intestine constituting the intestinal immune system and reacting with the bone marrow cells with the proliferation factor obtained.

First, the small intestine of L3 (lipopolysaccharide) -independent C3H / He mice is collected and the Peyer's patch of the small intestinal wall is cut and transferred to a cold RPMI-1640 medium containing petridish. After placing the metal body on the patch (mesh # 100) and releasing the cells from the patch while pressing with a syringe rubber stopper, the cell suspension is filtered using the metal body (mesh # 200) and washed three times with the medium. Adjust the cell concentration to 2 × 106 cells / mL RPMI-1640, divide 200 μL into 96 well plates, and incubate in a 5% CO2 incubator at 37 ° C for 5 days to recover the supernatant. Use as culture supernatant. Inject the bone marrow cells while injecting RPMI-1640 medium from the shin bone of the same mouse, filter and wash as described above, and adjust to 2.5 × 105 cells / mL RPMI-1640. Proliferation of bone marrow cells was measured using Alamar Blue Reduction assay. After dispensing 100 μL of the adjusted bone marrow cells into 96 well plates, the cells were cultured from Peyer's patch cells and 50 μL of RPMI-1640 medium for 6 days at 37 ° C. Incubate. 5 μL of Alamar Blue solution was added 5 hours before the end of the incubation, and the fluorescence intensity was measured using Fluoroskan II at excitation 544 nm and emission 590 nm. The myeloid cell proliferation promoting activity of the sample quantifies the proliferation of myeloid cells from the difference from the control.

Figure 4 is a graph showing the effect of myeloid cell proliferation activity through Peyer's patch on the hydrothermal extracts of the experimental group kimchi and control kimchi.

Referring to FIG. 4, the bone marrow cell proliferation activity as an indicator for activating the circulatory immune response by stimulating Peyer's patch cells, which are local immune tissues distributed in the small intestine, was examined. Kimchi group (Experimental group 7 days: 1.11 times, Experimental group 15 days: 1.17 times) showed a slightly higher activity than the saline control group without kimchi (Saline Control), 7 days, unlike mitogen and macrophages The activity increased slightly in the samples that had been aged for 15 days, indicating that the related active substances were different, but the functional activity of the kimchi hot water extract was different from that of Kimchi Control (1.17 times the control). Otherwise it was shown to have reduced or only similar levels of activity.

 c. Macrophage Proliferation Activity

Macrophage activity was determined by measuring macrophage lysosomal phosphatase activity.

That is, after injecting 1 mL of thioglycollate medium into the abdominal cavity of 5-10 week old male ICR mouse, 48-72 hours later, the mouse was intraperitoneally washed with RPMI-1640 medium and the macrophages were recovered from the abdominal cavity. The recovered macrophages are washed twice with RPMI-1640 medium and redispersed in RPMI-1640 medium so that the cell number is 1 × 10 6 cells / mL RPMI-1640. 180 μL of this dispersion is dispensed into each well of a 96-well plate and incubated for 2 hours at 37 ° C. in a 5% CO 2 incubator, whereby macrophage cells adhere to the base wall of each well plate to form a monolayer. After two hours, the non-adherent cells were washed and removed, and 180 μL of RPMI-1640 medium containing 10% fetal bovine serum (FBS) was added to each well, and 20 μL of the sample was added to each cell at 37 ° C. in a 5% CO2 incubator. Redeploy for 24 hours to activate macrophages. 0.1% triton X-100 (25 μL) was added to the activated macrophage monolayer to dissolve the macrophage cell membrane, and 100mM p-nitrophenyl phosphate (150μL) and 0.1M citrate buffer (50μL) were used as substrates for the lysosome phosphatase. ) And react in acidic condition for 1 hour, then stop the reaction by adding 0.2M borate buffer and measure the absorbance at 405 nm with ELISA reader to measure the activity of phophatase.

Hot water extracts were prepared from them over 7 days and 15 days while the experimental group kimchi was aged at 4 ° C. That is, each kimchi was crushed using a total amount homogenizer, water was added to the sample including the liquid and stirred, and the supernatant was lyophilized to prepare hot water extract of kimchi.

The proliferative activity of macrophages recovered by injection of thioglycollate medium into the ICR abdominal cavity was measured by lysosomal enzyme activity. As shown in FIG. Even after 7 days (experimental group 7 days) 1.24 times after 15 days (experimental group 15 days) showed 1.16 times more active than saline control without the addition of kimchi (Saline Control) significantly more active Increased, but decreased over time.

These trends were similar in the mitogen, but there was no significant difference, but the optimal ripening time, like the mitogen, seems to be suitable for one week, and the addition of black garlic and brown rice mycelium showed that the functionalities were reinforced. see.

Ha. Anticancer activity

  The toxic effects of the samples on cancer cell lines (MCF7 (breast cancer), NCI-H460 (lung cancer), MKN45 (stomach cancer), HCT116 (colon cancer)) were measured by MTT assay method. , Incubated at 37 ° C and 5% CO2 in RPMI medium 1640 medium containing 1% antibiotics, and cultured cancer cells were completely separated from T-flask bottom by adding trypsin / EDTA (1mM) solution and reused in RPMI medium. Harm. Adjust the cell solution concentration so that the cell count is 1.5 × 104 / mL (NCI-H460), 2 × 104 / mL (MCF7, HCT116) to 5 × 104 / mL (MKN45), and the sample is immediately filtered through a 0.22 μm membrane filter. Sterilize and use in MTT assay. Cisplatin (0.5 mg / mL) was used as a positive control and the cytotoxic effect of the sample was calculated by the following equation.

Cytotoxic activity (%) = (1- (A550Sample ÷ A550nagative control)) × 100

Table 13 shows the results of preparing hot water extracts of functional kimchi prepared with black garlic and brown rice mycelium and their toxic activity against cancer cell lines.

Referring to Table 13, the anticancer activity of breast water and lung cancer cell lines of the hot water extract prepared from the experimental group Kimchi 7 or 15 days after the production of kimchi did not show a significant difference in activity compared to the conventional kimchi. However, the hot water extracts prepared from kimchi aged after 7 days showed increased anticancer activity in gastric cancer and colorectal cancer cell lines compared to the control group kimchi hot water extracts.

From these results, it was confirmed that the anticancer activity of functional kimchi added with black garlic and brown rice mycelium showed excellent cancer cell suppression effect in gastric cancer and colorectal cancer, thereby enhancing the physiological activity as functional kimchi. In addition, it was found that the optimum ripening time of the kimchi of the experimental group was one week.

TABLE 13

As described above, Kimchi for enhanced physiological activity containing black garlic and situational mycelium powder according to the present invention by adding together black garlic and situational mycelium powder, physiological activities such as antioxidant, immunity and anticancer activity are enhanced, and also in preference It did not degrade compared to conventional kimchi.

In the above description, the embodiments of the present invention have been described. However, various changes and modifications can be made by those skilled in the art. Such changes and modifications may belong to the present invention without departing from the scope of the present invention. Therefore, the scope of the present invention will be determined by the claims described below.

Figure 1 shows the change of pH according to the storage period of the experimental group kimchi added black garlic and situation mushroom mycelium powder.

Figure 2 shows the change in acidity according to the storage period of the experimental group kimchi and control kimchi.

Figure 3 is a graph showing the effect of mitogen activity on the hot water extracts of the experimental group kimchi and control kimchi.

Figure 4 is a graph showing the effect of myeloid cell proliferation activity through Peyer's patch on the hydrothermal extracts of the experimental group kimchi and control kimchi.

5 is a graph showing the effect of macrophage activity on the hot water extracts of the experimental group kimchi and the control kimchi.

Claims (3)

Preparation step of seasoning beef to add black garlic and situation mycelia powder; And In the method of producing bioactive strengthening kimchi comprising the step of producing kimchi using the seasoning, The physiologically enhanced kimchi has a higher flavonoid content and polyphenol content than black garlic and ground mycelium powder-free kimchi, Method for producing physiologically-enhanced kimchi, characterized in that it has the ability to inhibit the formation of lipid peroxide according to kimchi storage. The method of claim 1, The situation mycelium powder is a method for producing physiologically active kimchi, characterized in that the brown rice microbial powder. The method of claim 1, The seasoning beef, pickled radish, red pepper powder, white sugar, black garlic, brown rice mycelium powder, pear, buttercup, salted shrimp, onion, ginger, chives, L-glutamate sodium, sesame oil, sesame or 5'-ribonucleotide disodium Method for producing physiologically enhanced kimchi, characterized in that.

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