KR100714408B1 - Method for manufacturing of red ginseng bacillus - Google Patents

Abstract

The present invention relates to a method for producing red ginseng Bacillus subtilis bacteria to be added to red ginseng soybean paste, red ginseng soy sauce, red ginseng pepper paste, red ginseng cheongukjang, specifically, boiled soybeans and cooled by laver soaking in a colander to overlap each other at 42-45 ° C. The first fermentation step is put into a fermentation chamber maintaining the temperature for 48 hours

After cooling the fermented product extracted from the fermentation room to 10 ℃ and put into a screw crusher and mixing the red ginseng powder prepared while crushing

Secondary fermentation step of fermenting red ginseng powder mixed in colander for 25 hours in fermentation chamber maintaining 35 ~ 38 ℃, drying step of fermenting secondary fermented product at 55 ~ 60 ℃ and grinding mixture A method of producing Bacillus subtilis bacteria, which is a mixture of red ginseng, obtained by sequentially pulverizing and powdering with red ginseng. It is effective in preventing bacteria, diarrhea, enteritis, diabetes and blood pressure, and can be used as various foods and functional food additives to obtain red ginseng bacillus subtilis bacillus, red ginseng soy sauce, red ginseng red pepper paste, red ginseng Cheonggukjang is to be made.

Red ginseng, Gochujang, Cheonggukjang, Bacillus subtilis

Description

Manufacturing method of Bacillus subtilis bacteria mixed with red ginseng {METHOD FOR MANUFACTURING OF RED GINSENG BACILLUS}

The present invention relates to a red ginseng soybean paste, red ginseng soy sauce, red ginseng red pepper paste, red ginseng to be added to the red ginseng soy sauce mixed bacillus subtilis bacteria.

Red ginseng refers to steaming or swelling ginseng or gelatinizing starch of ginseng by other means.

Red ginseng has a sedative and arousing effect on the central nervous system, and has been shown to be effective in preventing hypertension and arteriosclerosis by acting on the circulatory system.

It also reduces hematopoietic and blood sugar levels, protects the liver, and acts indirectly on sexual and reproductive effects by acting on the endocrine system. Anti-inflammatory and anti-tumor effects (抗 腫 瘍 作用) has a protective effect against radiation, it is also known to soften the skin.

In addition, the most important of the effects of red ginseng is the adaptogen effect, which has been scientifically proven to increase the defense ability against various harmful effects stresses from the surrounding environment and to make the body adapt more easily.

Soy contains 41% protein. The amount of protein is the highest among crops, and the composition of amino acids is inferior to meat.

Lysine and tryptophan are essential in the soybeans because they contain a lot of essential nutrition than meat.

Soybeans can be used to prevent or treat arteriosclerosis, myocardial infarction, stroke and high blood pressure by lowering the amount of cholesterol in the blood.

Dietary fiber, polyunsaturated fatty acids (mainly nioleic acid; 51%), saponins, and trypsin inhibitors in soybeans are known to be important because they lower cholesterol levels or prevent cancer.

Lecithin in soybean oil is known to lower cholesterol and prevent senile dementia.

On the other hand, air has about 20mg / 100g of vitamin E, which is effective for beauty and anti-aging.

Korea developed soybean culture by developing fermented foods using soybeans. Meju is made from raw soybeans harvested in October, and the beans are boiled. Although the shape of the meju varies, it is generally molded into 8 * 12 * 120cm ³

Traditionally, dried shaped meju is dried, tied with melon and tied to the eaves to ferment naturally.

Meju fermentation is an important process for making miso.

Bacillus subtilis bacteria (chocobacteria) are attached to the spores, and after the beans have been boiled, they are also the main spontaneous fermentation bacteria. It is a fermented bacterium and other bacteria, such as mold, are also involved in fermentation to produce traditional meju.

Interestingly, the fermented meju in the laboratory study significantly improved cancer prevention and anticancer effects. Soybean meju, which has a larger surface area than traditional sake meju, was more effective.

Looking at the data that have been studied microorganisms until now, the reality is that the interest is attracting, such as applying for a patent focusing on the type of microorganism strain or newly discovered type.

But if we want to develop more than penicillin, we need to be more concerned about what the host is when it is strongly stimulated or when it is cultured.

Cordyceps is a phenomenon in which the microorganisms that have been flying in the air attach to insects and take effect from insects in winter and grow like mushrooms in summer, and there are 7-8 kinds of insects that are hosts of cordyceps.

Among them, you can easily see silkworm cordyceps.

Grapes have both the host and the strain at the same time because microorganisms that have been flying in the air settle on the surface of the grapes.

In addition to making soybeans by fermenting soybeans with meju, making wheat into yeast, and making sake, ginseng becomes red ginseng and jihwang sulfur, which is a blood-reducing function.

But all of these have different effects depending on the host or the conditions under which the microorganism grows, rather than the type of microorganism.

In light of this fact, the type of microorganism is important, but I think it is more important to study the environment and nutritional host.

Recently, long-fermented soybean paste had been researched for its anti-cancer effects. Meju and soybean paste are known to improve their cancer prevention function during soybean fermentation.

The present invention is to combine the soybean and red ginseng to achieve a more enhanced effect Bacillus subtilis bacteria first fermented in soybeans mixed with red ginseng and the second fermentation Basilu sub having a more enhanced function It is to make tilis by preparing the tilis bacteria, which will be described in more detail as follows.

1) Preparation

Prepare the same amount of beans and red ginseng powder.

2) manufacturing process

Boil soybeans, cool it, and put it in a colander to overlap each other, put it in a fermentation chamber maintaining a temperature of 42 ~ 45 ℃,

After the extraction from the fermentation chamber and cooled to 10 ℃ mixed into the screw crusher prepared by mixing the red ginseng powder and

Bacillus subtilis bacteria mixed with red ginseng powder in a colander and put into a fermentation chamber maintained at 35 ~ 38 ℃ and fermentation for 25 hours and

A drying step of drying the second fermented fermented product in a drying room at 55-60 ° C. until the water content is 13% by weight;

It is a manufacturing method obtained by sequentially performing the step of pulverizing the dried product to a size of 120 mesh by milling.

In the present invention prepared as described above, Bacillus subtilis bacteria involved in the fermentation process of soybeans are grown while being involved in fermentation by mixing with red ginseng powder to form red ginseng Bacillus subtilis bacteria having a more enhanced function, and miso, Soy sauce, red pepper paste, Cheonggukjang mixed with 10-15% by weight can be produced red ginseng miso, red ginseng soy sauce, red ginseng red pepper paste, red ginseng Cheonggukjang so that it can have an enhanced effect than the conventional Cheonggukjang.
Conventional Cheonggukjang also has the effect of preventing food poisoning, enteritis, bacterial diarrhea, constipation, diabetes, blood pressure, etc. It is known to have the effect of inhibiting DNA damage, but the degree is very weak, so the effect of Cheonggukjang is only 5 times improved. Even if this is a development value, the present invention has proved that it has an effect that exceeds it.

단일 청국장 분말의 항유전독성 효과
단일 청국장 분말의 항유전독성 효과를 측정하기 위해, 청국장 분말(가야원)을 사용하여 위와 동일한 방법으로 Comet assay를 실시하였다.
그 결과, 도 2에서 알 수 있는 바와 같이, 단일 청국장 분말의 항유전독성 효과는 단일 홍삼 분말의 경우와 비슷하게 나타났다. 즉, 청국장 추출물 원액의 DNA 손상 정도는 175로 홍삼보다 조금 더 탁월한 항유전독성 효과를 나타내었으며, 100배 희석액은 DNA 손상 정도가 247, 1000배 희석액은 DNA 손상 정도가 249로 거의 DNA 손상 억제 효과가 없는 것으로 나타났다.
(도 2)단일 청국장 분말의 항유전 독성 효과

홍삼 및 청국장 혼합 분말의 항유전독성 효과
홍삼 분말과 청국장 분말을 30:70(w/w)의 비로 혼합하여 혼합 분말 200 g을 얻은 뒤, 그 혼합 분말의 항유전독성 효과를 위에서와 동일한 방법으로 측정하였다.
그 결과, 도 3에 나타낸 바와 같이, 홍삼 분말과 청국장 분말을 혼합한 경우에는 상기 홍삼 분말과 청국장 분말을 각각 따로 분리하여 항유전독성 효과를 조사한 경우보다 그 효과가 훨씬 크다는 사실을 확인할 수 있었다.
즉, 홍삼 추출물 원액에서는 DNA 손상 정도가 187로 나타나고, 청국장 추출물 원액에서는 175로 나타난데 반해, 홍삼 및 청국장 혼합 추출물 원액에서는 155 내외로 현저히 적게 나타났으며, 상기 단일의 홍삼 추출물 및 청국장 추출물과 달리 홍삼 및 청국장의 혼합 추출물에서는 그 1000배 희석액에서도 DNA 손상 정도가 양성 대조구(250)보다 적은 214로 나타나, 충분한 정도의 DNA 손상 억제 효과가 있음을 알 수 있었다.
결론적으로, 본 발명의 홍삼 및 청국장의 혼합 분말은 1000배까지 희석된 상태로 식품에 첨가되어도 충분히 항유전독성을 나타낼 수 있을 것으로 기대되었다.
(도 3)홍삼 및 청국장 혼합 분말의 항유전독성 효과

홍삼 및 청국장 혼합 분말의 DNA 손상 복구 정도
홍삼, 청국장 그리고 홍삼 청국장 혼합 추출물을 분리하여 이들의 항유전독성 효과를 서로 조사 비교하였다.
도 4에서 볼 수 있는 대로 홍삼 청국장 혼합 추출물의 항유전독성 효과는 크게 나타났으며 단독으로 사용하는 것 보다 혼합 추출물은 synergistic한 효과가 나타나는 것으로 보여진다.
DNA 손상 억제는 도 5에서 보여지는 바와 같이 홍삼 청국장 추출물 원액에서는 37%의 억제 효과가 있는 것으로 나타났으며 10배 희석 액에서는 20% 억제 효과가 있으며 100배 희석 액에는 16% 그리고 1000배 희석 액에는 14%로 항유전독성 효과가 있는 것으로 나타났다.
(도 4)홍삼 청국장의 항유전독성 효과 비교

(도 5)DNA 손상 복구 정도

The present invention was made to test the effect of the red ginseng Cheonggukjang made by using the red ginseng bacillus subtilis bacteria to confirm this effect and to see the effect on the human body.
In this experiment, the antigenetic toxicity (DNA damage inhibition) effect according to the dilution ratio of red ginseng and Cheonggukjang mixed powder was compared with the antigenetic toxicity effect of single red ginseng powder and single Cheonggukjang powder using Comet assay. And by checking the anti-genetic toxicity effect according to the mixing ratio of the Cheonggukjang mixed powder, to determine the preferred mixing ratio, and to prepare food compositions with different addition ratios of the red ginseng and Cheonggukjang mixed powder, to test the anti-genetic toxicity effect of the food compositions and The objective of the invention was achieved by performing a sensory evaluation to confirm the proper addition ratio.
As a result, the food additive of the present invention composed of a mixture of red ginseng and Cheonggukjang showed much better DNA damage inhibition effect than a single red ginseng powder and a single Cheonggukjang powder.
In order to investigate the antigenetic effects, to detect genotoxic substances such as carcinogens that cause strand breaks in DNA, damaged DNA is stained with DNA staining reagent and observed under fluorescence microscope for accurate and rapid analysis. Single cell electrophoresis, that is, Comet assay was used.
MNNG (N-Methyl-N'-nitro-N-nitrosoguanidine), which is a direct carcinogen used in the present invention, is purchased from Fluka, Germany, and refrigerated at 4 ° C. The cell line used in the present invention is mouse M. Non-tumoral nomal 3T3 cells (ATCC CCL No. 163), which are not malignant tumors of broth (mouse embryo) origin, are available from Korea Cell Line Bank.
For culture and preservation of cell lines, DMEM (Dulbecco's modified Eagle's mediument, Sigma USA) was charged with calf serum (Gibco, USA) 10, penicillin G (sigma, USA) 100 units / mL, streptomycin 100 μl / mL of streptomycin sulfate (sigma, USA) is added, and a wet culture apparatus (Labline instrument, USA) maintains 37 ° C. and 5% CO 2 in a 10 cm diameter round tissue culture dish (Falcon, USA). Culture was carried out), and the cells were passaged twice a week before reaching confluent, discarded after 20 passages, and replaced with new cells.
(Experiment result)
Antigenotoxic Effects of Single Red Ginseng Powder
In order to measure the anti-genetic toxicity of the single red ginseng powder, first, the red ginseng powder (Punggi ginseng Corp.) was dissolved in distilled water for 24 hours and centrifuged to obtain a supernatant. The supernatant was filtered three times, and the resulting solution was frozen in a deep freezer for 24 hours and then lyophilized to obtain red ginseng concentrate powder. 200 g of this powder was dissolved in 1 L of distilled water to prepare a stock solution. The stock solution was diluted 10 times by mixing 100 µl of the stock solution and 900 µl of distilled water. In the same manner, dilutions were obtained 100 and 1000 times to obtain respective dilutions.
The cell line used in this experiment was used 3T3 cells, and the initial inoculation of the 3T3 cells to 1 × 105 cells were incubated for 48 hours at 37 ℃, 5% CO2 wet incubator to investigate the DNA damage inhibition effect. At this time, a cell treated with only MNNG was used as a positive control.
First, 1 mg of MNNG, a carcinogen to 3T3 cells, was dissolved in 5 ml of PBS solution, and then treated with 10 μl of each treatment to damage DNA of 3T3 cells, followed by the red ginseng extract solution, 10-fold dilution, 100-fold dilution, and 1000. 10 µl each of the pear dilutions was mixed with HBSS to determine the extent of DNA damage recovery through the Comet assay. Before observing the effect of inhibiting DNA damage, the 3T3 cells were first immersed in a mixed solution of 125 ml of alkaline buffer, 25 ml of DMSO and 2.5 ml of Triton-x, and refrigerated for 1 hour. The membranes were dissolved in NaCl and EDTA. After 20 minutes pre-reaction in the electrophoresis buffer, electrophoresis was performed for 20 minutes at 300mA, 25V. Subsequently, the cells were immersed in a neutralization buffer for 5 minutes and neutralized. Then, the handles of the slides were dried, and the dye was diluted 1: 1 with PBS in a fluorescent dye reagent YOYO-1 to examine the degree of DNA damage. (Molecular probes, USA) stained with 75ul. Stained slides were observed at 250 × magnification on a fluorescence microscope (CSB-FEI, China).
As a result, as shown in Figure 1, the DNA damage in the positive control was 250, but the DNA damage in the red ginseng extract undiluted solution was 187, showing that the red ginseng extract has an excellent antigenetic effect. On the other hand, in the 100-fold and 1000-fold dilutions of red ginseng extract, the degree of DNA damage was similar to that of the positive control of 242 and 248, respectively.
(FIG. 1) Antigenotoxic Effects of Single Red Ginseng Powder

Antigenotoxic Effects of Single Cheonggukjang Powder
In order to measure the anti-genetic toxicity of a single Cheonggukjang powder, a Comet assay was performed using the Cheonggukjang powder (Kayawon) in the same manner as above.
As a result, as can be seen in Figure 2, the anti-genetic effect of the single Cheonggukjang powder appeared similar to the case of a single red ginseng powder. In other words, the DNA damage of the Cheonggukjang extract was 175, which was more excellent than the red ginseng. The 100-fold dilution was 247, and the 1000-fold dilution was 249. Appeared to be absent.
(FIG. 2) Antigenetic Toxicity Effect of Single Cheonggukjang Powder

Antigenotoxic Effects of Red Ginseng and Cheonggukjang Mixtures
Red ginseng powder and Cheonggukjang powder were mixed in a ratio of 30:70 (w / w) to obtain 200 g of mixed powder, and then the antigenetic effect of the mixed powder was measured in the same manner as above.
As a result, as shown in Figure 3, when the red ginseng powder and Cheonggukjang powder was mixed, it was confirmed that the effect is much greater than the case of separately separating the red ginseng powder and Cheonggukjang powder to investigate the anti-genetic effects.
In other words, the amount of DNA damage was 187 in the red ginseng extract and 175 in the chungkukjang extract, whereas the red ginseng and the Cheonggukjang extracts were markedly less than 155 in the mixed extract of the red ginseng extract. In the mixed extract of red ginseng and Cheonggukjang, the DNA damage was 214 less than that of the positive control 250 even in the 1000-fold dilution, indicating that the DNA damage was sufficiently inhibited.
In conclusion, the mixed powder of red ginseng and Cheonggukjang of the present invention was expected to exhibit sufficient anti-genetic toxicity even when added to foods diluted up to 1000 times.
(FIG. 3) Antigenotoxic effect of red ginseng and Cheonggukjang mixed powder

DNA Damage Recovery Level of Red Ginseng and Cheonggukjang Mix Powder
Red ginseng, Cheonggukjang and red ginseng Cheonggukjang mixed extracts were isolated and their anti-genetic effects were investigated.
As shown in Figure 4, the anti-genetic toxicity effect of the red ginseng Cheonggukjang mixed extract was shown to be large, and the mixed extract was shown to have a synergistic effect than used alone.
As shown in FIG. 5, inhibition of DNA damage was shown to have 37% inhibitory effect in the red ginseng chungkukjang extract undiluted solution, 20% inhibitory effect in 10-fold dilutions, and 16% and 1000-fold dilutions in 100-fold dilutions. Had antigenotoxic effect of 14%.
(Figure 4) Comparison of anti-genetic toxicity effect of red ginseng Cheonggukjang

(Figure 5) DNA damage recovery degree

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Claims (1)

Boil the soybeans and send out a laver, put them in a colander, sprinkle with each other, put them in a fermentation chamber maintaining a temperature of 43 ~ 45 ℃, and ferment for 48 hours. After cooling the fermented soybeans from the fermentation room to 10 ℃ and put into a screw crusher and mixing the prepared red ginseng powder and crushed The second fermentation step is to put the crushed product mixed with red ginseng powder in a colander and fermented for 25 hours in a fermentation chamber maintained at 35 ~ 38 ℃ and Method for producing Bacillus subtilis bacteria mixed with red ginseng obtained by sequentially performing the step of drying the secondary fermentation at 55 ~ 60 ℃ and the step of powdering the dried product