KR101567635B1 - Manufacture Method of Functional Korean Traditional Soybean Paste comprising Codyceps militaris and its Extract - Google Patents

Abstract

The present invention relates to a method for preparing soy sauce and soy sauce having a health-healing function to which cordisepin is added at a high content of caterpillar fungus. More specifically, the present invention relates to a method for producing soy sauce and soy sauce, And a method for producing soy sauce and soybean paste having modern palatability and functionality. According to the present invention, soy sauce and doenjang added with Chinese cabbage extract having immunity enhancement, anti-cancer and liver function improving ability according to the present invention can be utilized as high value-added products as they have inherent value and various functions.

Description

Technical Field [0001] The present invention relates to a method for producing a functional soybean paste containing millitary cordyceps and an extract thereof,

The present invention relates to Cordyceps, which contains a high amount of cordycepin and is excellent in immunity enhancement, cancer prevention effect and liver function enhancement effect, and a new method for producing the same using the extract.

As the dietary life worsens due to the improvement of the living standards due to the growth of the economy and the society, a variety of lifestyle-related diseases such as cardiovascular diseases, diabetes, circulatory diseases such as hypertension, and cancer are occurring. As the onset of lifestyle-related diseases is closely related to the diet, the improvement of dietary life and interest in functional foods are emerging.

Since ancient times, soybean paste and soy sauce are fermented soybeans, and they are the most important seasoning food in Korea. These traditional soybean fermented foods are prepared by boiling soybeans in autumn and then forming meju into cubes and hanging them with rice straw. As Meju is dried, microorganisms suitable for proteolysis grow and contain a large amount of proteolytic enzymes secreted by the produced cells. The prepared meju is added with salt, water, etc. and aged for several months, it becomes taste and flavor as soy sauce and miso. These doenjang and soy sauce are excellent nutritional foods as essential amino acid and fatty acid sources that are lacking in our diet with a grain oriented diet. They are good nutritional foods, Is known as a functional food having activity.

Cordyceps is one of the insect parasitic fungi that form fruiting bodies in pupa and larva by transferring to larva or summer grass in winter. It is a kind of mushroom that grows as a nutrient source of insect protein source when spore is transplanted into insect body. The Cordyceps mushroom strain belongs to the genus Cordyceps belonging to the Ascycycetes and Clavicipitaceae, and 300 species of 100 genera are known worldwide including Korea, China, and Japan. Most of them are parasitic on insects and produce fruiting bodies on insect bodies that become host. The host insects may be parasitic to the spider in addition to the lepidoptera (Cordyceps militaris), C. periwinkle (C. sobolifera), C. sphecocephala, other species of beetles, grasshoppers, Among them, Cordyceps militaris used in the present invention is one of traditional medicines widely used in China and has been widely used as medicines and tonic foods in the West Asia region. Cordycepsin (cordycepin), adenosine, carotenoid, carotenoid, and polysaccharide are the functional ingredients of Cordyceps. Among them, codisepin is a typical functional ingredient, which is a structure in which the hydroxyl group at the 3 'position of adenosine, which is very similar to deoxyadenosine, which is one of the constituent substances of DNA, is eliminated and has a competitive action with the constituent components of genes, It is known that it exhibits various pharmacological activities such as inflammation, antioxidation, immune enhancement and anti-platelet function. Cordyceps containing this highly functional codepin has been mainly used for health promotion and has been used for anti-inflammatory, anti-cancer, Have been reported.

Recently, with the interest in health, various functionalities of traditional foods have been reported, leading to an increase in consumption of fermented foods in conjunction with a well-being trend. Therefore, in order to add diversity of traditional foods, functional foods and vegetable materials have been recently added to develop functional foods and foods that satisfy consumers' preferences. Therefore, various kinds of ingredients such as soy sauce and soybean paste, which are the most basic seasoning foods of Korea, And flavored soy sauce and soy sauce are in the spotlight.

The present inventors have completed the present invention by studying a method for fermenting soy sauce and miso with healing function for human health by using fruiting bodies and extracts of Cordyceps mushroom which are excellent in immunity enhancement, anticancer and liver function recovery.

Korea Patent No. 10-0944636

Thus, the inventors of the present invention have completed the present invention by confirming that it is possible to produce a high-content functional juice of cordisepin by producing a juice using a Millitary Cordyceps sinensis strain.

Therefore, an object of the present invention is to provide a method for producing soy sauce, which comprises the steps of increasing and crushing soybeans in order to produce functional soy sauce excellent in immunity, anticancer and liver function recovery, Which comprises fermenting a fermented product by adding a hot-water extract, and filtering and separating the fermented product.

It is another object of the present invention to provide a high-content functional codissein produced by the above process.

In order to achieve the above object, the present invention provides a method for producing soy sauce, comprising the steps of increasing and crushing soybeans, adding corn filed powder to pulverized soybeans, shaping into meju, fermenting fermented soybeans with salt water and Cordyceps wax extract, And a step of isolating the plant.

In one embodiment of the present invention, the Cordyceps sinensis may be C. militaris JLM 0636.

In one embodiment of the present invention, the caterpillar fungus powder is obtained by pulverizing caterpillars of the caterpillar fungus, and the caterpillar fungus extract is prepared by adding 500 to 1500 parts by weight of distilled water to 100 parts by weight of cordyceps, It may be a hot-water extract extracted for 3 hours.

In one embodiment of the present invention, the meju forming step may be performed by adding 0.5 to 7.5 parts by weight of cordierose powder to 100 parts by weight of ground soybean.

In one embodiment of the present invention, the fermentation may be performed by adding 10 to 20 parts by weight of salt water and 2.5 to 15 parts by weight of caterpillar fungus extract to 100 parts by weight of Meju.

In one embodiment of the present invention, the soy sauce may be soy sauce or soy sauce.

In one embodiment of the present invention, the soy sauce is prepared by adding 5 to 75% by volume of Cordyceps horticultural extract at a concentration of 1 to 10% (w / v) based on the volume of the filtrate obtained by filtering the fermented product, .

In an embodiment of the present invention, the soybean paste may be prepared by adding 5 to 10 parts by weight of a powdery mildew of a mushroom powder to 100 parts by weight of a solid obtained by filtering the fermented product, followed by fermentation.

The present invention also provides a high-content functional codissein produced by the above process.

In one embodiment of the present invention, the herbarium may be one having anti-inflammation, immunity enhancement, liver function improvement and anticancer activity.

In one embodiment of the present invention, the functional soybean may be soy sauce or soybean paste.

In one embodiment of the present invention, the soybean paste contains Cordycepin at 0.20 to 0.50 ml / ml, and the soy sauce contains 0.4 to 1.20 mg / g of cordycepin. There may be something.

The Cordisceps militaris JLM 0636 fruiting body and Cordyceps sinensis extract of the present invention have a high content of intrinsic functional ingredients of Cordyceps sinensis. Accordingly, the functional soy sauce of the present invention containing it as an active ingredient can be usefully used as a functional seasoning, because it has the advantage of modern Chinese health by adding functionalities of Cordyceps, while satisfying taste and taste of modern people.

Also, the doenjang or soy sauce according to the present invention contains a large amount of cordycepin, carotenoid, polysaccharide and the like and is a food having an excellent function in anti-inflammatory action, immunity enhancement, improvement of liver function, prevention of cancer or therapeutic effect of cancer.

Figure 1 shows an HPLC graph showing the content of cordycepin in the high content cordycepin cordyceps
2 is a graph showing the anti-inflammatory immunoactivity of cordisepin
3 is a graph showing a mechanism of inducing apoptosis (suicide inducing mechanism) of human prostate cancer cells by cordycepin
FIG. 4 shows the effect of improving the liver function of cordisepin high-content Cordyceps sinensis
Fig. 5 shows a process for producing high-content Cordyceps sinensis-containing meju, soy sauce, and doenjang
6 is a graph showing the results of sensory evaluation of soy sauce and doenjang containing cordisepin high content ciliothermophilus

The terms used in the present invention are defined as follows.

Throughout this specification, unless the context requires otherwise, the words "comprises" and "comprising " are to be understood as encompassing the stated step or material, but not the exclusion of any other step or material .

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications referred to herein are incorporated herein by reference.

The present invention relates to a functional soybean prepared by a conventional method having excellent functionality in anti-inflammatory action, immunity enhancement, liver function improvement, cancer prevention or cancer treatment, which comprises a large amount of cordycepin, carotenoid, polysaccharide, The present invention relates to a method for producing soybean paste such as soybean paste and soybean paste, which can maximize the effect of the extract by using a powder of Cordyceps sinensis fruit and an extract of Cordyceps sinensis fruit in order to increase the effective components of cordisepine, carotenoid, polysaccharide and the like.

In addition, the present invention may be applied to the production of soybean paste such as kochujang, soy sauce, soybean paste, chonggukjang, chunjang, ssamjang, and hyochukjang using a mycelia of funguscoptera or fruiting body, a culture medium containing funguscoptera mycelium or fruiting body.

The Cordyceps sinensis strains of the present invention may be selected from the group consisting of Cordyceps sinensis, Cordyceps militaris, Cordyceps scarabaecola, Cordyceps nutans, Such as Cordyceps sphecocephala, Cordyceps tricentri, Paecilomyceps tenuipes, Phytocordyceps ninchukiospora, Paecilomyces japonica, and the like It is also possible to use all kinds of similarly known cordyceps.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

< Example  1>

Preparation of Cordyceps sinensis powder and extract

The high content of cordisepin caterpillars (C. militaris JLM 0636) used in this experiment was supplied from Cheongwon Agricultural Products (Gimhae, Kyungsangnamdo). The powdery mildew of Cordyceps mushroom was prepared by pulverizing the dried mushroom of Cordyceps sinensis with a pulverizer. The extracted mushroom was pulverized, and 10 times of distilled water was added to the weight of the pulverized material. The extract was subjected to hot water extraction at 121 ° C for 3 hours. The extracts were filtered and concentrated 10 times.

< Example  2>

Determination of cordine content in Cordyceps

To measure the content of cordycepin contained in cordyceps used in Example 1, the present inventors added pulverized cordyceps to distilled water 10 times the weight of the pulverized material, extracted at 121 ° C for 3 hours, filtered and centrifuged (8,000 rpm), and the resulting supernatant was lyophilized. The content of cordycepin was calculated by the ratio of cordycepin peak area in the chromatography obtained by adding distilled water to the lyophilized product and performing HPLC. The calibration curve was prepared using a commercially available cordycepin standard.

The results of analysis of the content of cordycepin from the extracts obtained by hot water extraction of Cordyceps were shown in FIG. 1, and the content of C. militaris (JLM0636) was 7.42 ± 0.10 mg / g in terms of dry weight. In the case of C. militaris (other), the content of 1.01 ± 0.37 mg / g was higher than that of commercially available Cordyceps, and the cordycepin contained in Cordyceps was not decomposed at high temperature and pressure .

< Example  3>

Preparation of Meju, Soy Sauce and Doenjang Using Cordyceps and Their Extracts

The process for preparing meju, soy sauce, and doenjang using Cordyceps sinensis and its extract according to the present invention is shown in the schematic diagram of Fig. More specifically, soybean (soybean) is soaked at a temperature of 25 ° C for 10 hours by adding 10 times as much water as soybean. Add the so-called beans to the beaker and boil it for 3 hours at 121 ° C. Cool, and grind with a grinder to prepare. To the pulverized soybeans, 5% of the powdery mushroom powder (pulverized product) prepared in Example 1 was added, mixed with a stirrer, and molded into a square shape to prepare meju with Cordyceps.

The prepared meju was dried at 30 ° C for 3 days and then fermented on dried rice straw for 80 days. The fermented meju was thoroughly washed with running water and then the water was removed and placed in a jar. The extract was prepared by adding 20% of brine of 10 times of the weight of the fermented meju and 30% of the extract of 10% After adding vol%, charcoal and red pepper were floated and fermented for 120 days. After fermentation, the meju lumps were recovered and used as a raw material of miso. The filtrate was separated by filtration and used as raw material for soy sauce. That is, 10% of Cordyceps sinensis powder (crushed) was added to meju lumps, and the mixture was fermented for 60 days to prepare doenjang containing Cordyceps. In addition, the filtrate, which is the raw material of the soy sauce, was aged for 90 days in a jar to prepare a Chinese cabbage-containing soy sauce. At this time, 10% of the Chinese cabbage extract was added to the filtrate at 20 vol% based on the volume of the filtrate as the soy sauce raw material. Soy sauce and doenjang were prepared by the above-described method, except that the Chinese cabbage was not added as the control group.

< Experimental Example  1>

Analysis of anti-inflammatory and immunological activity of cordisepin

The inventors of the present invention conducted the following experiment to analyze whether cordisepin, which is a useful physiologically active ingredient, has excellent anti-inflammatory and immunological activity, in order to produce functional soybean paste.

Immunoreactivity was measured by intraperitoneal macrophages isolated from peritoneal cavity of C57 / BL6 mice treated with LPS, which is an inflammatory inducer, and cordysepin, which is a useful physiologically active ingredient, to final concentrations of 2 μg / ml and 10 μg / ml, respectively 37 ° C, 5% CO ₂ And cultured for 6, 24, and 48 hours, respectively. The cytokines IL-1β, IL-6, IL-12 and TNF-α secreted from the culture medium were measured using an ELISA MAX ™ Deluxe set kit.

Macrophages exposed to LPS secrete proinflammatory cytokines such as IL-1β, IL-12, IL-6, and TNF-α to mediate early inflammatory responses to outbreaks of microorganisms, It acts to probe external factors. The phagocyte is removed by macrophages and is involved in the activation of innate immunity and adaptive immune system by various cytokines secreted from activated macrophages. In particular, macrophages, which are antigen presenting cells, T lymphocytes recognize the antigen.

As a result, as shown in FIG. 2, in the case of the group treated with Codisepin, the immunoreactivity was enhanced as compared with that in the group treated with LPS alone, and in particular, Codisepin was induced in the mouse peritoneal macrophages by LPS In the inflammatory response, the production of IL-1β, an early inflammation-mediated cytokine, is induced to efficiently induce the immune response, thereby activating the immune response. The conversion from the innate immunity to adaptive immunity induced by natural immune cells such as macrophages T lymphocytes, IL-12 and TNF-α secreted more than LPS-stimulated Codisceptin, indicating that Codisepin is more likely to be produced by macrophages than T-lymphocytes As shown in Fig. In addition, IL-6 production by IL-6 was significantly increased by LPS treatment alone, but after 48 hours, IL-6 production by IL- And the production of LPS was increased more than that of LPS alone. This suggests that activation of B lymphocytes mainly in the late phase of immune response induces an increase of antibody - induced immune response in the late phase of treatment with cordisepin.

< Experimental Example  2>

Of prostate cancer cells by codedine Apoptosis  ( Apoptosis : Cell suicide mechanism ) Induction analysis

In order to confirm that cell proliferation inhibition by cordycepin is related to apoptosis in human prostate cancer cell line PC3, PC3 cells were treated with concentration of cordisepin for 24 hours at each concentration, and sub-G1 phase measurement and DNA fragmentation were performed through FACs .

As a result, an increase in the frequency of sub-G1 group (Figs. 3A and 3B) corresponding to the frequency of apoptosis-induced cell cycle distribution (Fig. 3A and 3B) and agarose gel electrophoresis By confirming the fragmentation of DNA (Fig. 3C), it was confirmed that the suppression of PC3 cell proliferation by cordisepin was closely related to induction of apoptosis.

Therefore, the inventors of the present invention have found that the codisepin component activates the immune response and has anticancer activity at the same time.

< Experimental Example  3>

Analysis of liver function improvement effect of Cordyceps sinensis

The following experiments were conducted to determine if the codedine component has the effect of improving liver function.

Sprague-Dawley mice were divided into the following groups: normal (N), alcohol-treated control (C), alcohol and Paecilomyces japonica (CPJ), alcohol and Cordyceps militaris (CCM), Alcohol and Cordyceps militaris (JLM0636) (CCMα), Alcohol and Silymarin group (CSM), and fed to mice. After that, ADH activity was measured from the liver tissues of each mouse group by Bergmeyer's method, and histopathological observation of liver tissue was observed by optical microscope after hematozarin and eosin (H & E) staining.

For reference, alcoholic degradation in the liver by chronic alcohol consumption is first induced by the conversion of alcohol to acetaldehyde by alcohol dehydrogenase (ADH) and cytochrome P-450. Acetaldehyde accumulated in liver tissue is affected by ADH and ALDH activity. ADH activity increases and ALDH activity decreases when chronic alcohol is administered to experimental animals.

As a result, ADH activity in hepatic tissue was slightly increased in alcohol control group compared with normal group, but activity was significantly increased by administration of CPJ, CCM and CCMα. The ALDH activity was decreased in the alcohol control group as compared with the normal group, and the activity was increased in the CPJ, CCM, and CCMα administration groups (FIG. 4A). These results indicate that Cordyceps sinensis promotes alcohol metabolism enzymatic ADH and ALDH activities in liver tissues and reduces the alcohol and acetaldehyde concentrations in blood. Especially, it shows that ADH and ALDH activities are highest in CCMα - treated group.

The accumulation of lipid in the liver cells is the most common symptom in the early stage of alcoholic liver toxicity. As a result of histological examination of the liver tissue, the liver lobular structure was well maintained around the blood vessels in the control group, Rich cytoplasmic cytoplasm and round nucleus, and hepatocyte plate was mostly maintained in one layer. In the CPJ, CCM, and CCMα administration groups, it was similar to that of the normal group, and it was confirmed that the cordisepin high content millitary caterpillar fungus inhibited the hepatocyte protective effect . &Lt; / RTI &gt;

< Experimental Example  4>

Determination of the content of cordycepin in soy sauce and doenjang prepared by the method of the present invention

The contents of cordisepin, which is a useful physiologically active ingredient, in the liver and doenjang of the present invention using the caterpillar fungus produced in the above Examples were respectively confirmed. The prepared soybean paste was diluted with distilled water of 10 times the weight of doenjang, and the diluted solution was centrifuged. The supernatant was used as a sample. The soy sauce was prepared by diluting 5 times of the volume of the soy sauce filtrate with distilled water. And each was calculated by HPLC by area ratio of codistepin peak. For quantification, a calibration curve was prepared using a commercially available codistepin standard.

As a result of the analysis, in the liver prepared by the method of the present invention, the content of cordycepin was 0.81 ± 0.37 mg / ml and that of soybean paste was 0.35 ± 0.12 mg / g. . In addition, no control was detected in soy sauce and doenjang without control (see Table 2).

Therefore, the inventors of the present invention have found that the method for producing soy sauce and soybean paste according to the present invention can produce a functional soybean soup without deteriorating the effective physiologically active ingredient codisepine, while maintaining useful activity.

< Experimental Example  5>

Analysis of general components of soy sauce and doenjang prepared by the method of the present invention

The general compositional analysis of the liverworts and doenjang prepared by the method of the present invention was confirmed by measuring the pH, the sugar content, and the salinity. The salinity was measured using a salinity meter (PAL-03S, ATAGO, Japan). The pH of the sample was measured with a pH meter (Twin pH, Japan) (PAL-1, ATAGO, Japan) were measured.

The difference in pH, sugar content and salinity of soy sauce and doenjang were investigated when caterpillar fungus was added (Table 3, 4). As a result, pH was 4.8 in pH and pH was 4.8 in the unfermented soy sauce and pH 4.9 in the non - additive soy sauce. The pH value of doenjang and doenjang doenjang added with doenjang .

The changes of salinity were found to be 6.1% and 6.5 in soy sauce added with and without added cucumber herb, respectively, and 3.8% and 4.6% in doenjang added with doenjang and doenjang added, respectively. The salinity of soy sauce was affected by the concentration of initial salinity. In case of doenjang, salinity value was low due to the mixing of mycelia of. The sugar content of cucumber - added soy sauce and non - added soy sauce were 4.5 brix and 4.7 brix, respectively. The addition of doenjang and doenjang doenjang showed 3.0 brix and 2.7 brix, respectively. It was confirmed that the sugar content was slightly higher when added to Cordyceps. However, these results suggest that the addition of Cordyceps has a slight change in sugar content. From these results, it was confirmed that there was no difference in pH, sugar content, and salinity in the additions of Cordyceps mellifera and without added soy sauce.

< Experimental Example  6>

Sensory evaluation of soy sauce and miso prepared by the method of the present invention

The sensory evaluation of soy sauce and doenjang added with Chinese caterpillar fungus was confirmed by taste, flavor, color, texture and overall acceptability. The sensory evaluation of soy sauce and doenjang added with Chinese caterpillar fungicide was conducted by Dong - A university between 30 and 30 years old, and fermented soy sauce and doenjang were evaluated by color, taste, aroma and overall preference in 7 items And subjected to statistical processing with a SAS program (see FIG. 6).

The results of sensory evaluation showed that the color and flavor were superior to those of soy sauce and doenjang by adding flavor and flavor of Chinese caterpillar fungus. Therefore, it was highly judged when the Chinese cabbage was added as a whole. As a result of the final evaluation, soy sauce and doenjang added with Chinese caterpillar fungus were highly evaluated in color, taste, aroma and overall acceptability.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (12)

As a method for producing soy sauce or soybean paste using caterpillar fungus,
The method
Growing and crushing soybeans;
Adding the powdered caterpillar fungus powder to the pulverized soybeans and shaping them into meju;
Fermenting Mechu with salt water and Cordyceps wax hydrothermal extract; And
Filtering and separating the fermentation product,
The method for producing soy sauce or soybean paste as set forth in claim 1, wherein the Chinese cabbage is JLM 0636. delete The method according to claim 1,
The above-mentioned caterpillar fungus powder is obtained by pulverizing caterpillar fungus body, and the above-mentioned caterpillar fungus extract is a hot-water extract obtained by adding 500 to 1500 parts by weight of distilled water to 100 parts by weight of caterpillar fungus extract at 90 to 140 ° C for 1 to 3 hours Of soy sauce or soybean paste using caterpillar fungus. The method according to claim 1,
The method for preparing soybean paste or soybean paste according to claim 1, wherein the meju-forming step comprises adding 0.5 to 7.5 parts by weight of cordierite powder to 100 parts by weight of pulverized soybean. The method according to claim 1,
Wherein the fermentation is performed by adding 10-20 parts by weight of salt water and 2.5 parts by weight to 15 parts by weight of caterpillar fungus hydrothermal extract to 100 parts by weight of Meju, and fermenting the fermented soybean paste or soy sauce using the caterpillar fungus. delete The method according to claim 1,
Wherein the soy sauce is prepared by adding 5 to 75% by volume of Cordyceps mizugi hot water extract at a concentration of 1 to 10% (w / v) based on the volume of the filtrate obtained by filtering the fermented product,
Wherein the soybean paste is prepared by adding 5 to 10 parts by weight of powdery mildew powder to 100 parts by weight of solid matter obtained by filtering the fermented product, and then fermenting the mixture to produce soy sauce or soybean paste. delete delete delete delete delete

Images