KR100945670B1 - Composition of healthy food containing fermented soybean powder - Google Patents

Abstract

The present invention relates to a health food composition comprising the Cheonggukjang powder, and more specifically, it is composed of Cheonggukjang powder, soybean hot water extract fermentation broth powder, ginseng extract powder and grain flour excellent health enhancement food, antioxidant activity and antithrombotic activity It relates to a composition. Ingestion of the health food composition of the present invention has an immune enhancing, antioxidant and antithrombotic effect.

Cheonggukjang Powder, Soybean Hot Water Extract Fermentation Powder, Immune Enhancement, Antioxidant, Antithrombotic

Description

Composition of healthy food containing fermented soybean powder             

1 is a graph showing the immune enhancing activity according to the preparation method of the Chungkookjang powder.

Figure 2 is a graph showing the effect of inhibiting the production of peroxide according to the production method of Chungkookjang powder.

Figure 3 is a graph showing the immune enhancing activity according to the type of soybean inoculation strains.

Figure 4 is a graph showing the immune enhancing activity of soy hot water extract fermentation broth prepared using Bacillus pumilus JB-1 (Bacillus pumilus KCTC 10461BP) according to the present invention.

5 is a graph showing the antithrombotic activity of the Isaac extract methanol extract according to the present invention.

6 is a graph showing the immunopotentiating activity of cereal flour according to the present invention.

7 is a graph showing the immune enhancing activity of the health food composition according to the present invention.

8 is a graph showing the immuno-enhancing activity according to the content of soy hot water extract fermentation broth powder of the health food composition according to the present invention.                 

9 is a graph showing the DPPH radical scavenging activity of the health food composition according to the present invention.

10 is a graph showing the peroxide production inhibitory activity of the health food composition according to the present invention.

11 is a graph showing the DNA damage inhibiting ability of the health food composition according to the present invention.

12 is a graph showing the antithrombotic activity of the health food composition according to the present invention.

The present invention relates to a health food composition comprising the Cheonggukjang powder, and more specifically, it is composed of Cheonggukjang powder, soybean hot water extract fermentation broth powder, ginseng extract powder and grain flour excellent health enhancement food, antioxidant activity and antithrombotic activity It relates to a composition.

Cheonggukjang is a representative fermented food of Korea, which is made from soybean. Cheonggukjang is made by fermenting soybean with Bacillus subtilis. During fermentation, enzymes produced by Bacillus subtilis produce its unique taste and scent and are derived from sugar and protein of raw soybean. It is a high-quality fermented food with high protein and fat content due to the generation of viscous substances, a mixture of levan form fructan and polyglutamate.

In recent years, many researchers have studied isoflavone, phytic acid, saponin, trypsin inhibitor, tocopherol, unsaturated fatty acid, dietary fiber, oligosaccharide, γ-glu It has been found that various bioactive substances such as tamiltranspeptidase (γ-glutamyltranspeptidase, γ-GTP), antioxidants and thrombolytic enzymes are contained in large amounts.

However, despite the various functionalities of Cheonggukjang, the development of processed food using Cheonggukjang is insufficient.

The inventors of the present invention, while studying processed foods using Cheonggukjang, by adding a variety of grains to the grain flour mixed with the Cheonggukjang powder, soybean hot water extract fermentation powder, the ginseng extract powder has an immune enhancing activity, antioxidant activity and antithrombotic activity The present invention has been completed by preparing a functional food composition.

Accordingly, an object of the present invention is to provide a health food composition consisting of 5-30% by weight of Cheonggukjang powder, 0.01-0.5% by weight of soybean hot water extract fermentation broth powder, 0.01-0.2% by weight ginseng extract powder and 69.5-94.5% by weight grains will be.

In order to achieve the above object, the present invention is composed of 5-30% by weight of Cheonggukjang powder, 0.01-0.5% by weight of soybean hot water extract fermentation broth powder, 0.01-0.2% by weight ginseng extract powder and 69.5-94.5% by weight flour Provide a food composition.

The health food composition according to the present invention is characterized by having immunopotentiating activity, antioxidant activity or antithrombotic activity.

Hereinafter, the present invention will be described in detail.

The health food composition according to the present invention is prepared by adding functional raw materials, such as Cheonggukjang powder, soybean hot water extract fermentation broth, and Isaac extract extract powder, which have various physiological activities to grains mixed with various grains. It has very good activity or antithrombotic activity.

Cheonggukjang contains an enzyme called nattokinase, which is produced by Bacillus and is known for its excellent thrombolytic ability, genistein, which is known to have anticancer effects, isoflavones, which are known to be similar to estrogen, and which acts to lower blood cholesterol. Vegetable cholesterol, melanoidin having strong antioxidant activity and anticancer activity, polyglutammate, and polymer nucleic acid having anticancer activity and immunostimulating activity are included.

The inventors of the present invention was prepared in the form of Cheonggukjang powder to prepare a health food composition using the Cheonggukjang containing various bioactive substances as described above. In an experimental example of the present invention, the cheongukjang was hot-air dried or lyophilized, and then pulverized to prepare each powder, and the immuno-enhancing activity and the inhibition of peroxide generation according to the drying method were examined. It was confirmed that the enhanced activity and the peroxide generation inhibitory activity is excellent (see Experimental Example 1 and Experimental Example 2).

Therefore, it is preferable that the Cheonggukjang powder used in the present invention is prepared by lyophilizing and drying the Cheonggukjang. Cheonggukjang may be prepared by a conventional method of inoculating Bacillus subtilis with fermented soybean. Lyophilization of the Cheonggukjang can be easily performed using a freeze-drying device. Preferably, lyophilization is performed for 48-72 hours so that the moisture content is 8-10% or less. After lyophilization is completed, Cheonggukjang is made into powder using a grinder. The powder is preferably 30 mesh to 50 mesh.

Functional materials of soybean are known as dietary fiber, oligosaccharides, isoflavonoids, phytic acid, trypsin inhibitors, saponins, proteins, vegetable sterols and phenolic compounds. These soybeans are fermented rather than eaten, enriching essential amino acids, vitamins, pantothenic acid and various enzymes. In particular, a viscous substance having an anticancer effect, a polymer nucleic acid having an immune enhancing activity, an antioxidant, and the like are produced.

The soybean hot water extract fermentation broth powder used in the present invention may be prepared by inoculating a fermentation broth obtained by inoculating Bacillus sp. Strain into the soybean hot water extract discharged from the steaming process of Chungkookjang soybeans after freeze drying and grinding. . The Bacillus genus strain is preferably Bacillus pumilus JB-1 ( Bacillus pumilus KCTC 10461BP). In an embodiment of the present invention, soybean hot water extract fermentation powder powder is prepared by lyophilizing the fermentation broth obtained by diluting soybean hot water extract with distilled water 10 times, inoculating Bacillus pumilus JB-1, and incubating for 48 hours. It was used as a functional raw material of the health food composition according to the present invention (see Example 1).

In order to select a strain suitable for the fermentation of soybean hot water extract, the present inventors suspend and dilute commercial Cheonggukjang with sterile distilled water, and separate the colonies having excellent growth by spreading the dilutions on LB solid medium and then cultivating them. JB-1, JB-2, and JB-3 are named according to their differences. JB-1, JB-2 and JB-3 isolated from Cheonggukjang and known Bacillus subtilis, yeast, and lactic acid bacteria were inoculated into soybean hydrothermal extract and fermented to obtain a fermentation broth, followed by immuno-enhancing activity of the fermentation broth. As a result of the measurement, the immunopotentiation activity of the fermentation broth when inoculated with JB-1 was found to be superior to that of other strains (Experimental Example 3). Thus, the present inventors were able to confirm that Bacillus pumilus as a result of identifying the JB-1 strain. Accordingly, the present inventors named the JB-1 strain as Bacillus pumilus JB-1, and deposited the deposit number KCTC 10461BP with the Korea Gene Bank on April 9, 2003.

In addition, the present inventors inoculated Bacillus pumilus JB-1 showing the excellent immune enhancing effect in soybean hydrothermal extract solution and then cultured by measuring the immunopotentiation activity according to the concentration of the culture solution obtained, low concentration (0.03mg / ml), it was confirmed that the immune enhancing activity of about 200% (see Experimental Example 4).

Persicaria filiforme NAKAI is a perennial herb that grows on mountain streams or forest edges throughout the country. It is used as a medicine for skin diseases, heart disease, etc. in the folk villages, and young sprouts are known to be edible. However, no antithrombotic activity of Isaac has been reported to date.

The inventors of the present invention examined the antithrombotic activity of the methanol extract, and found that the antithrombotic activity was very excellent in a concentration-dependent manner (see Experimental Example 5). Therefore, the crushed methanol powder was used as a functional raw material of the health food composition according to the present invention.

The ginseng powder used in the composition according to the present invention can be obtained by extracting ginseng alcohol with an alcohol according to a known method to obtain an extract and then completely removing the alcohol in the extract liquid. The alcohol used may be ethanol or methanol, preferably methanol. In an embodiment of the present invention, the ginseng was dried in the shade, then chopped and extracted with methanol to obtain a methanol extract, which was concentrated under reduced pressure in a 90 ° C. water bath to completely remove methanol, and then pulverized to obtain ginseng powder.

The flour used in the health food of the present invention is in the form of a mixture of various grains. Preferably, 10-30% by weight of glutinous rice, 10-30% by weight of brown rice, 10-30% by weight of barley rice, 5-15% by weight of weak beans, 1-10% by weight of white beans, 1-10% by weight of black radish, black sesame 1- 10 wt%, perilla 1-5 wt%. Carbohydrate: lipid: protein calorie intake ratio of 60-48%: 23-30%: 18-22% in the case of the addition of Cheonggukjang after mixing the flour in the above preferred ratio, the carbohydrate: lipid: protein of It can be said that the formulation is close to 65%: 20%: 15%. In addition, the perilla is an excellent source of ω-3 fatty acid, ω-3 fatty acid occupies 63.1% of the fat content of 39.5%, the intake of ω-3 fatty acids is greatly increased when ingested in the compounding ratio of the present invention have. The grain flour may be prepared by mixing glutinous rice, brown rice, barley rice, weak soybeans, white baekju, yulmu, black sesame seeds and perilla on low heat, pulverized to prepare a powder, and then mixing them in the composition ratio. As a result of examining the immunopotentiation activity of the grain flour, there was no immunopotentiation activity (see Experimental Example 6). Thus, in the present invention, grain flour is used as an energy source for supplying carbohydrates, lipids and proteins.

The health food composition according to the present invention can be prepared by mixing the above functional raw materials in a predetermined ratio to the grains prepared as described above.

Preferably it is mixed in the ratio of 5-30% by weight Cheonggukjang powder, 0.01-0.5% by weight soybean hot water extract fermentation powder, 0.01-0.2% by weight ginseng extract powder and 69.5-94.5% by weight flour.

At this time, the immune enhancing activity of the Cheonggukjang powder added to the health food composition of the present invention is concentration-dependent (Fig. 7) can be expected to increase the immune activity concentration-dependent more the amount added, but more than 30% by weight of the Cheonggukjang powder If added, the characteristic odor becomes stronger and the palatability is lowered. If it is added below 5% by weight, it is difficult to expect at least 200% of the immune enhancing effect, so it is preferable to mix the Cheonggukjang powder to contain 5-30% by weight. Do.

In addition, the immune enhancing activity of soy hot water extract fermentation broth showed an increase of 200% even at very low concentrations (0.03 mg / ml), and the immune enhancing activity did not show a proportional increase even when the concentration was increased (FIG. 4). On the other hand, when the soybean hot water extract fermentation broth powder is added to the health food composition according to the present invention at 0.5% by weight, the immune enhancing activity increases depending on the concentration of the water extract of the health food composition, but the immunological activity increases at a certain concentration. Did not do it. When the soy hot water extract fermentation broth was added at 1.0% by weight, the immune activity was increased when the concentration of the water extract of the health food composition was low (0.3 mg / ml), but it was decreased when the concentration was high. (FIG. 8). Therefore, the preferred mixing ratio of soy hot water extract fermentation broth powder was determined to 0.01 to 0.5% by weight.

Cereal meal has no effect on immune activity (Fig. 6), so it is formulated to consume 65%: 20%: 15% energy ratio of carbohydrate: fat: protein, which is a desirable intake ratio of energy nutrients in consideration of nutrition. It was.

The antithrombotic activity of aspirin used as a positive control was 25-30% of that of Isaac, and aspirin-level anti-aspirin was recommended when aspirin was used to prevent the recurrence of heart disease. The anti-thrombotic activity was included in one dose, and if the antithrombotic activity is too strong in the body, it can be said that the prevention of heart disease-related diseases, but the risk of bleeding is relatively strong, so the daily intake was determined to be 0.01 to 0.2% by weight.

As a result of investigating the functionality of the health food composition according to the present invention, the inventors were able to confirm that immunity enhancement, antioxidant and antithrombotic activity were very excellent (see Experimental Example 7).

The health food composition prepared as described above has the advantage of having a small smell peculiar to Cheonggukjang, good shelf life, easy and convenient ingestion anywhere, and easy storage.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1>

Preparation of a health food composition comprising the Chungkukjang powder according to the present invention

After fermentation for two days, the non-seasoned Cheonggukjang was freeze-dried at -70 ° C using a lyophilizer (Samwon Freeze Dry System, SFDSF24, Samwon co.) And then pulverized with a grinder to prepare a 40-mesh Cheonggukjang powder.

To prepare soybean hot water extract fermentation broth powder, soybean hot water extract discharged during boiling of soybeans was recovered and diluted 10-fold with distilled water. The diluted solution was inoculated with Bacillus pumilus JB-1 ( Bacillus pumilus KCTC 10461BP) isolated from Cheonggukjang, and then sodium hydroxide (NaOH) was added to adjust the pH to 8 and incubated at 37 ° C. for 48 hours. When the culture was completed, the fermentation broth was collected, lyophilized in the same manner as above, and then pulverized with a grinder to prepare soybean hot water extract fermentation broth powder.

The ginseng extract powder was dried after dipping the ginseng extract, immersed in methanol, and then immersed in methanol three times for three days to remove the methanol completely by concentrating under reduced pressure in a water bath at 90 ° C. to prepare a grind powder.

Grain flour used in the present invention is glutinous rice, brown rice, barley rice, medicinal soybeans, white tae, jukmu, black sesame and perilla, each lightly roasted and then pulverized to prepare a powder, the grain powder 20% by weight, brown rice 20% %, 20% by weight of barley, 12% by weight of soybeans, 8% by weight of white beans, 8% by weight of radish, 8% by weight of black sesame and 4% by weight of perilla.

Each of the ingredients prepared by the above method was mixed in a combination as shown in Table 1 to prepare a health food composition according to the present invention.

material Contains 10% of Cheonggukjang 20% of Cheonggukjang Compounding ratio (%) g / 1 serving (50 g) Compounding ratio (%) g / 1 serving (50 g) Flour 89.45 44.725 79.45 39.725 Cheonggukjang Powder 10.0 5.0 20.0 10.0 Soybean Fermented Extract Powder 0.5 0.25 0.5 0.25 Ginseng extract powder 0.05 0.025 0.05 0.025 Sum 100.0 50.0 100.0 50.0

Experimental Example 1

Investigation of Immune Enhancing Activity According to Manufacturing Method of Cheonggukjang Powder

After fermentation for two days, the undiluted Cheonggukjang is lyophilized or hot air dried and then pulverized with a grinder, 10 times the weight of distilled water is added, followed by extraction at 50 ° C. for 2 hours, followed by centrifugation (8000 rpm, 30 minutes). After lyophilization and pulverized was used as a sample. Lyophilization was carried out in the same manner as in Example 1, hot air drying was dried for 12 hours at 150 ℃ in a hot air dryer. The immuno-enhancing activity of lyophilized powder and hot air dried powder of Cheonggukjang prepared by the above method was investigated by comparing mouse splenocyte proliferation rate. Isolation of mouse splenocytes was performed by known methods (Mishell et al., Selected methods in cellular immunology, WH Freeman and Co. p4-27, 1980). After sacrificing the mouse by the method of cervical dislocation, spleen cells were released, and then erythrocytes were removed with Lysing buffer (Tris-buffered ammonium chloride 0.87%, pH 7.2) and DMEM containing 10% FBS (fetal bovine serum) The culture was adjusted to 5 × 10 ⁶ cells / ml. 90 μl per well was put into a 96-well plate, and 10 μl of the Cheonggukjang powder sample diluted to different concentrations were dispensed, respectively, and the plate was incubated for 48 hours in a 37 ° C., 5% CO ₂ incubator. 50 μl of MTT solution (1 mg / ml) was added thereto, followed by further incubation for 4 hours, the supernatant was removed, and 150 μl of DMSO was added thereto, and left for 5 minutes. Then, the absorbance was measured at 580 nm using an ELISA reader. Proliferation rate of mouse splenocytes was calculated using the following equation.

Proliferation% = (absorbance of sample / absorbance of control) x 100 ... (1)

As a result, the immuno-enhancing activity of the Cheonggukjang powder was about 50% of that of the lyophilized powder. Therefore, it was found that the immunological activity of the lyophilized powder of Cheonggukjang was better (FIG. 1).

Experimental Example 2

Investigation of the Peroxide Production Inhibition Effect by the Preparation Method of Cheonggukjang Powder

The inhibitory effect of the peroxide production of the hot air dried powder and lyophilized powder of Cheonggukjang prepared in the same manner as in Experimental Example 1 was investigated. 50 μl of 10% homogenate (10% homogenate) of mouse liver and Cheonggukjang hot-air dried powder or lyophilized powder samples diluted by concentration were mixed, and then 10 mM ascorbic acid and 0.5 mM FeSO 4 were added as oxidants at 37 ° C. Shaking incubation for 60 minutes, immersed in ice to terminate the reaction, and 100 μl of the reaction solution was taken, using known methods (Ohkawa et al., Anal. Biochem., 95, 351-358, 1979) to TBARS (thiobarbituric acid). The amount of rective substanc es) was measured at 532 nm. Peroxide production inhibition rate (%) of the sample was calculated using Equation 2 below, and the concentration (IC ₅₀ ) where the absorbance when the sample was added indicates 50% of the control absorbance was calculated.

Peroxide formation inhibition rate (%) = [1- (absorbance of sample / absorbance of control)] × 100 ... (2)

As a result, peroxide production inhibition rate of lyophilized powder of Cheonggukjang was higher than that of hot air dried powder. In addition, the IC ₅₀ value of the lyophilized powder was 1.2 mg / ml and the IC ₅₀ value of the hot air dried powder was 4.3 mg / ml (FIG. 2). Therefore, it was found that the effect of inhibiting the peroxide generation of the lyophilized powder of Cheonggukjang was much better than that of the hot air dried powder.

Experimental Example 3

Selection of Fermentation Strains Used to Prepare Soy Hot Water Extract Fermentation Solution

Bacillus subtilis known as representative strains of strains JB-1, JB-2 and JB-3, Cheongguk coli isolated from Cheonggukjang by the present inventors to select fermentation strains used to prepare fermentation broth of soy hot water extract . subtilis KCTC 1914), Lactic acid bacteria L. casei KCTC 3189, Lactobacillus vulgaris ( L. bulgaris KCTC 3188) and baker's yeast ( S. cerevisiae IFO 0233), respectively. The immunopotentiating activity according to the strain used was investigated.

The JB-1, JB-2 and JB-3 are diluted with 1 g of commercial Cheonggukjang after suspension with sterile distilled water, and the diluted solution is smeared on an LB solid medium (Difco) and incubated at 30 ° C. for 2 days to provide excellent growth. Secured by separating colonies.

Using each strain of the above, the fermentation is completed in crushed Cheonggukjang with a grinder, and added 10 times the volume of water and stirred for 2 hours at 50 ℃ centrifugation (8000rpm, 30 minutes) to take the supernatant and freeze-dried The immunopotentiating activity was examined in the same manner as in Experimental Example 1.

As a result, the inoculation activity of JB-1, a strain isolated from Cheonggukjang, was the highest as about 410%, which is more than 250% higher than other strains. JB-2 and JB-3 isolated from Cheonggukjang showed 163% and 158% activity, respectively. In the case of Bacillus subtilis, immunopotentiating activity was 152%, similar to that of JB-2 and JB-3 (FIG. 3). Therefore, the JB-1 was selected as a fermentation strain to be used in the preparation of soybean hot water extract fermentation broth powder according to the present invention.

The JB-1 strain was identified as Bcillus pumilus by biolog strain identification experiment (GP2 Microplate, Biolog, CA, USA) and membrane fatty acid composition analysis (Gas chromatograohy HP 5890, USA). The JB-1 strain was named Bacillus pumilus JB-1 and was deposited on April 9, 2003, with the Korean Gene Bank under accession number KCTC 10461BP.

Experimental Example 4

Investigation of Immune Enhancing Activity of Soy Hot Water Extract Fermentation Solution Using Bacillus Fumilus JB-1

Using Bacillus pumilus JB-1 (KCTC 10461BP) to prepare a soybean hot water extract fermentation broth by the method of Example 1 and the immuno-enhancing effect of the concentration was investigated in the same manner as in Experiment 1. At this time, the soybean hot water extract fermentation broth was prepared using yeast ( S. cerevisiae IFO 0233), and compared with the immune enhancing activity when the Bacillus pumilus JB-1 was used.

As a result of the experiment, when the fermentation broth of soybean hot water extract was prepared using yeast, there was almost no immunopotentiating activity. On the other hand, using Bacillus pumilus JB-1 showed an immunostimulatory activity of about 200% even at a low concentration of 0.03mg / ml. However, when the concentration of soy hot water extract fermentation broth was 0.03 mg / ml or more, the immune enhancing activity did not show a proportional increase (FIG. 4).

Experimental Example 5

Investigation of Antithrombotic Activity of Extracts

The antithrombotic activity of the ear extract methanol extract prepared in the same manner as in Example 1 was investigated. Antithrombotic activity was examined by measuring the inhibitory activity against thrombin by blood coagulation time. In a test tube, 50 µl of 0.02 M CaCl ₂ solution, 50 µl of bovine thrombin (0.5 U) as a blood coagulation enzyme, and 10 µl of a methanol extract were mixed. 100 μl of plasma isolated from blood treated with sodium citrate was added thereto, and the time of blood coagulation was measured using an Amelung Coagulom eter KC 1A. As a positive control, aspirin was used instead of the methanol extract.

Experimental results showed that the thrombin time when the methanol extract was not added was 29 seconds, but the concentration increased depending on the case where the methanol extract was added, and the thrombin concentration was 1 mg / ml. The time was over 300 seconds, indicating an increase in activity of at least 1000%. In addition, when compared to the antithrombotic activity of aspirin, which is prescribed at low concentrations to prevent recurrence by inhibiting thrombogenesis in heart disease patients, it was found to have more than three times stronger activity (FIG. 5).

Experimental Example 6

Investigation of Immune Enhancement Activity of Wheat Flour

In order to investigate the immuno-enhancing activity of the flour of Example 1, 10 times the volume of the grain weight of water was added and stirred at 50 ° C. for 2 hours, and then the supernatant was treated in the same manner as the method of preparing the water extract. Was prepared. Immunostimulatory activity was investigated in the same manner as in Experimental Example 1.

As a result of the experiment, in the case of grain flour, the immune enhancing activity did not appear (FIG. 6).

Experimental Example 7

Investigation of Immune Enhancing Activity of Health Food Composition Containing Cheonggukjang Powder According to the Present Invention

7-1) Investigation of Immune Enhancing Activity According to Cheonggukjang Powder Content of the Health Food Composition of the Invention

The water extract was prepared by the method of Experimental Example 1 by adding 10 times the volume of water to the health food composition containing 10% by weight or 20% by weight of the cheongukjang powder of Example 1. Immunostimulating activity according to the concentration of the sample was investigated in the same manner as in Experimental Example 1.

As a result of the experiment, the higher the concentration of Cheonggukjang powder contained in the health food composition of the present invention, the higher the immune enhancing activity. In other words, the composition containing 20% by weight of Cheonggukjang powder showed an increase of 286%, and the composition containing 10% by weight of Cheonggukjang powder showed a 212% increase in activity (FIG. 7).

7-2) Investigation of immuno-enhancing activity according to soybean hot water extract fermentation broth powder content of health food composition of the present invention

The immuno-enhancing activity of the health food composition when 20% by weight of Cheonggukjang powder was added and 0% by weight, 0.5% by weight and 1.0% by weight of the soybean hot water extract fermentation broth were added were investigated in the same manner as in Experimental Example 1. At this time, the content of the other components added to the composition was the same as in Example 1. To this end, the water extract of the composition having a different soybean hot water extract fermentation broth powder content was prepared in the same manner as in Experimental Example 1.

As a result of the experiment, the soybean hydrothermal extract fermentation broth powder was added to increase the immune activity concentration-dependently. However, when the soy hot water extract fermentation broth powder was added at 1.0% by weight, the immune activity increased when the concentration of the water extract of the health food composition was low (0.3 mg / ml), but the immunoactivity decreased when the concentration was high. (FIG. 8).

Experimental Example 8

Investigation of Antioxidant Activity of Health Food Composition Containing Cheonggukjang Powder According to the Present Invention

Antioxidant activity of the health food composition according to the present invention was measured by examining DPPH radical scavenging ability, peroxide production inhibitory activity, DNA damage inhibitory activity.

8-1) DPPH Radical Scavenging Activity

The DPPH radical scavenging activity of the health food composition according to the present invention was investigated according to known methods (Hatano et al., Chem. Pharm. Bull ., 36, 2090-2097, 1988). To this end, methanol extract powder was prepared by the above method by adding 10 times the volume of methanol to the health food composition containing 10 wt% or 20 wt% of the Cheonggukjang powder of Example 1, and then diluting them to various concentrations, respectively, to prepare a sample. Was prepared. 50 μl of the sample was dispensed into a 96-well plate and 200 μl of 200 μM DPPH (diphenyl-2-picrylhydra zyl) solution was added per well. After reaction at 37 ° C. for 30 minutes, the absorbance at 515 nm was measured using an ELISA reader. The radical scavenging rate of the sample was calculated using the following equation.

Radical scavenging rate (%) = (absorbance of 1-sample / absorbance of control) × 100 ... (3)

As a result of the experiment, the higher the concentration of the Cheonggukjang powder contained in the health food composition of the present invention, the higher the DPPH radical scavenging activity. In other words, the composition containing 20 wt% of Chungkookjang powder showed a radical scavenging rate of 62.8% at 50mg / ml concentration and 86.8% at 100mg / ml concentration, and 38.3 respectively for the composition containing 10wt% of Chungkookjang powder. Clearance rates of% and 55.4% are shown (FIG. 9).

8-2) Inhibition of Peroxide Formation

Peroxide production inhibitory activity of the health food composition according to the present invention was investigated in the same manner as in Experimental Example 2. To this end, the water extract of the health food composition containing 10% by weight or 20% by weight of the cheongukjang powder of Example 1 was prepared in the same manner as in Experimental Example 7-1).

As a result of the experiment, the difference in the peroxide generation inhibitory ability was not significant according to the weight percent of Chungkookjang powder contained in the health food composition of the present invention, and IC ₅₀ value was 9.37 mg / ml and 20 weight for the composition containing 10 wt% of Chungkookjang powder. The composition contained% was 6.76 mg / ml (FIG. 10).

8-3) DNA Damage Inhibition Investigation

The DNA damage inhibiting ability of the health food composition according to the present invention was investigated by a known method (Singh et al., Exp. Cell Res ., 175, 184-191, 1988). To this end, the water extract of the health food composition containing 20% by weight of the cheongukjang powder of Example 1 was prepared in the same manner as in Experimental Example 7-1) and then the final concentration was 50, 100 based on the weight of the health food composition, respectively. Samples were prepared by diluting to 200 μg / ml. After the sample was added to HepG2 cells, the cells were preincubated for 30 minutes and 10,000 cells were coated with 1% LMP agarose on a slide coated with 1% HMP agarose. It was left at 4 ° C. for 10 minutes and the slide was oxidized by soaking in 25 μM of H ₂ O ₂ for 5 minutes. At this time, the control group used a sample solvent instead of H ₂ O ₂ . The oxidized slides were treated with lysis buffer (pH 10) for 1 hour, unwinding buffer (pH 13) for 30 minutes, and then electrophoresed at 25V / 300mA for 25 minutes. After staining the cells with EtBr Comet image (Comet image) was measured by fluorescence microscopy (Leica, DM IL). The degree of DNA damage was expressed as a tail extent moment value using Equation 4 below. Two slides were made for each treatment and DNA images of 50 cells were analyzed for each slide.

Tail extent moment = tail length x tail% DNA ... (4)

As a result, DNA damage of cells treated with 20 wt% of Cheonggukjang powder was reduced in a concentration-dependent manner compared to cells treated only with H2O2. When the concentration of sample was 200 μg / ml, DNA damage was suppressed by 40%. (Figure 11).

Experimental Example 9

Investigation of antithrombotic activity of health food composition comprising Cheonggukjang powder according to the present invention

The antithrombotic activity of the health food composition of the present invention was investigated. To this end, after mixing the remaining powder material except for the ear is mixed in the same manner as in Example 1 to prepare a methanol extract and the methanol extract was diluted to a final concentration of 0.12, 0.24, 0.48 mg / ml by the method of Experimental Example 5 Antithrombotic activity was measured.

As a result, the antithrombotic activity in the case of the addition of Isaac was found to be high in a concentration-dependent manner (FIG. 12).

As described above, the health food composition comprising the Chungkukjang powder according to the present invention is prepared by adding functional raw materials such as Cheonggukjang powder to flour composed of various grains. There is.

Claims (6)

Health food composition consisting of 5-30% by weight of Cheonggukjang powder, 0.01-0.5% by weight soybean hot water extract fermentation broth powder, 0.01-0.2% by weight ginseng extract powder and 69.5-94.5% by weight grains. The health food composition of claim 1, wherein the Cheonggukjang powder is pulverized after freeze-drying the Cheonggukjang. The health food composition according to claim 1, wherein the soybean hot water extract fermentation broth is pulverized after lyophilizing the fermentation broth obtained by inoculating the soybean hot water extract by inoculating Bacillus sp. The health food composition of claim 3, wherein the strain of genus Bacillus is Bacillus pumilus KCTC 10461BP. According to claim 1, wherein the flour is 10-30% by weight glutinous rice, 10-30% by weight brown rice, 10-30% by weight barley rice, 5-15% by weight soybeans, 1-10% by weight white beans, 1-10% by weight Health food composition, characterized in that consisting of 1-10% by weight, black sesame 1-5% by weight. The health food composition according to any one of claims 1 to 5, wherein the composition has immunopotentiating activity, antioxidant activity or antithrombotic activity.

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