KR20200117320A - Vegetable Brown Sauce having high biological active materials and preparation method thereof - Google Patents

Abstract

Disclosed in the present invention are a vegetable brown sauce comprising: a pine mushroom mycelium fermented extract of mixed powder consisting of Toxicodendron vernicifluum, brown rice and soybeans; an extract of oriental medicinal herbs consisting of mixed ginseng of wood-cultivated ginseng and ginseng, Angelica gigas, Cynanchum wilfordii, Levisticum officinale, licorice and jujube; and fast-fermented bean paste stock, thereby containing a high content of physiologically active components of quercetin, epigallocatechin, protocatechuic acid and chlorogenic acid, and a method for preparing the same. The brown sauce according to the present invention has improved functions of antioxidant activity, diabetes amelioration and obesity amelioration.

Description

[Vegetable Brown Sauce having high biological active materials and preparation method thereof]

The present invention relates to a vegetable brown sauce with improved physiologically active ingredients and a method for preparing the same, and more particularly, to a fermented extract of matsutake mushroom mycelium of mixed powder consisting of sumac, brown rice and soybeans; Extracts of herbal medicinal herbs consisting of mixed ginseng of wild ginseng and ginseng, angelica, baeksuoh, cheonggung, licorice and jujube; And it is to provide a vegetable brown sauce containing a high content of physiologically active ingredients of quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid, including the source stock of cheonggukjang, and a method for producing the same.

As globalization makes it easier to access various Western foods, interest in Western-style recipes, especially sauces, is also increasing. In addition, the globalization of Korean food is accelerating the movement to develop or improve Korean traditional foods and dishes to suit Western tastes.

Brown sauce (sauce) is the most widely used sauce of Western meat that is used as a sauce for cooking after boiling with brown broth as the main ingredient and using an appropriate amount of spices and wine, then using a thickener to make concentration. Brown broth is prepared by boiling meat, bones, and vegetables for a certain period of time and concentrating. However, brown sauce mainly uses broth stock and does not meet the trend of reluctance to eat meat, so development of vegetable brown sauce that can replace it has been required.

Therefore, attempts have been made to prepare a sauce using cheonggukjang, but there was a problem that palatability was poor because the odor peculiar to cheonggukjang was not completely removed.In addition, as cheonggukjang is diluted a lot when making a sauce, functional and physiologically active substances derived from cheonggukjang It has become insignificant in the final source product, and improvement has been required.

Sumac ( Toxicodendron vernicifluum ) has been widely used as a medicinal agent because it helps digestion of the stomach, blood stagnation of the liver, and bleeding function of the heart. However, the use of lacquer is limited because it contains urushiol, which causes allergies or contact dermatitis such as blisters, itchiness, and rash. However, since lacquer tree contains various physiologically active substances other than urushiol, it exhibits various pharmacological actions, and attempts have been made to remove urushiol using green tea, mycelia of leaf mushrooms, or low-temperature extraction (Registration Patent No. 10-166139, 10-1182743 No., No. 10-1892617), a new removal technique is still required.

Quercetin, a type of flavonol, is known to have various pharmacological effects such as antioxidant, anti-inflammatory, antibacterial, antiviral, cancer cell proliferation inhibition, blood pressure lowering and capillary strengthening. In addition, epigallocatechin (epigallocatechin) is known to have effects such as lipase inhibitory activity and cardiovascular disease.

Protocatechuic acid, a kind of phenolic acid, is known for its antithrombotic activity, lipase inhibitory activity, and cardiovascular disease improvement activity, and chlorogenic acid is known for its antibacterial and anti-inflammatory properties.

Therefore, the development of a vegetable brown sauce containing a high content of flavonol and phenolic acid is desired.

Accordingly, the present inventors studied in order to meet the requirements of the prior art. As a result of the study, the fermented extract of matsutake mycelia of mixed powder consisting of lacquer, brown rice and soybeans, and mixed ginseng of wild ginseng and ginseng, angelica, baeksuoh, cheongoong, licorice and A vegetable brown sauce containing extracts of herbal medicinal herbs composed of jujube together with the source stock of Cheonggukjang contains a high content of physiologically active ingredients such as quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid, and has excellent functionality ( It was confirmed that it has an enhanced physiological activity) and the present invention was completed.

Therefore, an object of the present invention is a mixed powder consisting of lacquer, brown rice and soybean matsudake mycelium ( Tricholoma matsudake mycelium) ferment extract; Extracts of herbal medicinal herbs consisting of mixed ginseng of wild ginseng and ginseng, angelica, baeksuoh, cheonggung, licorice and jujube; And it includes a source stock of chungkukjang, quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid to provide a vegetable brown sauce having enhanced functionality (physiological activity) and a method of manufacturing the same .

In order to achieve the above object, the present invention is a matsutake mushroom mycelium fermented extract of mixed powder consisting of sumac, brown rice and soybeans; Extracts of herbal medicinal herbs consisting of mixed ginseng of wild ginseng and ginseng, angelica, baeksuoh, cheonggung, licorice and jujube; And cheonggukjang sauce stock, and the physiologically active ingredients of quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid are enhanced, and a vegetable brown sauce having excellent functionality is provided.

Lacquer mixed powder Pine mushroom mycelium Fermented extract

Brown sauce of the present invention comprises a fermented extract of pine mushroom mycelium of mixed powder consisting of sumac, brown rice and soybeans.

The mixed powder is preferably composed of lacquer powder 60-80% (w/w), brown rice powder 10-20% (w/w), and soybean powder 10-20% (w/w).

If sumac powder is less than 60%, the physiological activity may not be sufficient, and if it exceeds 80%, the amount of brown rice powder or soybean powder is small, which may delay the fermentation of pine mushroom mycelium and cause contamination. If the amount of brown rice powder and soybean powder, respectively, is less than 10%, fermentation may be delayed due to insufficient sugar and protein content required for fermentation of matsutake mycelium, and if it exceeds 20%, the amount of lacquer powder is insufficient and thus physiological activity is insufficient. May not.

After cutting, lacquer is sterilized by treating at 100~120℃ for 30~60 minutes, dried at 50~60℃ for 2~3 days, then chopped with a grinder and powdered.

In the present invention, fermentation is performed by adding water to the mixed powder so that it is about 50 to 60% moisture, swelling for about 1 to 2 hours, sterilizing at 100 to 120°C for 30 to 60 hours, and then adding 3 to 6% of matsutake mycelium ( v/w) and semi-solid fermentation.

When using pine mushroom mycelium in the present invention, enzyme activities such as cellulase, lignase, glucosidase, esterase, and protease are superior to other mushroom mycelium. As a result, it is advantageous not only to decompose urushiul, which is an allergen, but also to produce low-molecular physiologically active substances, and the unique scent of pine mushrooms can be added to the fermented product.

The fermented lacquer mixed powder according to the present invention prepared as described above was detoxified by removing urushiol, an allergen, and fermented into matsutake mycelium, rich in physiologically active substances, and matsutake mushroom Excellent palatability due to the added scent.

The extraction in the present invention is preferably ethanol extraction. Ethanol extraction can be extracted by adding 40 to 70% ethanol by 20 volume times and maintaining a 600 rpm speed at 30 to 40°C for 3 to 5 hours. Preferably, the extract is prepared by repeating extraction twice under the same conditions, filtering through filter paper, and adjusting the soluble solid content to 5 to 10 brix using a vacuum concentrator. If the soluble solid content is less than the above range, the physiological activity derived from the fermented lacquer tree mixed powder is not sufficient, and if it exceeds the above range, there is a problem in the palatability of the sauce due to bitter taste.

Extraction may be performed by mixing the fermented lacquer tree powder alone or with the following herbal medicine powder.

In the brown sauce of the present invention, the fermented extract of sumac mixed powder is added in an amount of 3 to 6% by volume when the total weight of the brown sauce is 100% by volume. If the amount is less than 3% by volume, physiological activity cannot be sufficiently expected, and if it is more than 6% by volume, problems in palatability such as bitter taste may occur.

Herbal medicine extract

The brown sauce of the present invention includes an extract of herbal medicinal material consisting of mixed ginseng of wild ginseng and ginseng, angelica, baeksuo, cheonggung, licorice and jujube.

Mixed ginseng is used by mixing wild ginseng and ginseng in a weight ratio of 1 to 2: 9 to 8. Mixed ginseng containing wild ginseng can supplement ginsenosides lacking in ginseng.

Angelica is known to have a blood-replenishing effect that replenishes blood, and its pharmacologically active ingredients include decursin, decursinol, umbelliferone, and β-sitosterol, which are coumarin derivatives.

Baeksu-o is known to be effective in improving the lipid metabolism of hyperlipidemia, as the tuber root is used as a herbal medicine in oriental medicine, and has the effect of preventing bone diseases including bone loss, inducing skeletal growth and the production of insulin-like growth factors, and improving lipid metabolism in hyperlipidemia. It contains 2,5-dihydroxyacetophenone and sinancon A.

Chungoong is a medicinal herb of the Angelicaaceae family. It contains various physiologically active ingredients, and has improved blood circulation, expanded blood vessels and increased blood flow by acting on the cardiovascular system, blood coagulation, central muscle relaxation, and anti-inflammatory effects.

Licorice is a medicinal herb of legumes, and its direct effect is to protect the stomach and neutralize toxicity. In particular, it works to neutralize the poison of other medicinal materials and alleviate the efficacy so that the efficacy is properly formulated.

Jujube contains various physiologically active ingredients such as betulinic acid to protect the stomach, anti-inflammatory and soothing.

Each of the herbal medicines is pulverized with a grinder and used as a powder.

The herbal medicinal material of the present invention includes mixed ginseng 15-25% (w/w), angelica 15-25% (w/w), baeksuo 15-25% (w/w), and cheonggung 15-25% (w/w) ), licorice 15-25% (w/w) and jujube 15-25% (w/w) are preferably included.

In the present invention, the extraction of the herbal medicinal material may be performed in the same manner as the extraction of the fermented lacquer mixed powder. After extracting the herbal medicinal material, the extract is prepared by adjusting the soluble solid content to 5-10 brix. If the soluble solid content is less than the above range, it is difficult to sufficiently expect physiological activity, and if it exceeds the above range, there is a problem in palatability of the sauce due to bitter taste and the like.

Likewise, extraction may be carried out by mixing oriental medicinal herbs alone or fermented lacquer mixed powder.

In the brown sauce of the present invention, the herbal medicine extract is added in an amount of 3 to 6% by volume when the total weight of the brown sauce is 100% by volume. If the amount is less than 3% by volume, physiological activity cannot be sufficiently expected, and if it is more than 6% by volume, problems in palatability such as bitter taste may occur.

Cheonggukjang sauce stock

Brown sauce of the present invention includes a stock of cheonggukjang sauce.

In the present invention, "sauce stock" means an undiluted solution that is the basis for the manufacture of sauce.

In the present invention, the source stock of cheonggukjang fermented with Bacillus amyloliquefaciens MGD02 strain is preferably used as the cheonggukjang sauce stock.

The present inventors isolated and identified Bacillus amyloliquefascience MGD02, which is the most suitable strain for the production of cheonggukjang sauce stock, and deposited with the Korea Agricultural Genetic Resource Center (KACC) on January 5, 2017 (accession number: KACC92158P). Cheonggukjang fermented with Bacillus amyloliquefascience MGD02 strain reduced its characteristic odor, resulting in excellent palatability and excellent storage stability.

Cheonggukjang sauce stock is dried Cheonggukjang fermented with Bacillus amyloliquefaciens MGD02 strain to prepare dried Cheonggukjang, then added 2 to 5 volume times of water to the dried Cheonggukjang, boiled, extracted, filtered, and concentrated to 20 to 40 brix And manufactured.

The cheonggukjang sauce stock according to the present invention has a reduced odor characteristic of cheonggukjang and has excellent storage stability.

In the brown sauce of the present invention, the cheonggukjang sauce stock is added in an amount of 50 to 60% by volume when the total weight of the brown sauce is 100% by volume. If the amount is less than 50% by volume, the flavor of the sauce due to the fermentation product is insufficient, and if it is more than 60% by volume, there is a problem that off-flavor of cheonggukjang may occur.

Brown sauce according to the present invention including the above extracts and cheonggukjeong sauce stock has physiologically active ingredients of quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid, respectively, at least 1,300 μg/ml and 2,600 μg/ml It was strengthened to more than, 400 µg/ml or more, and 490 µg/ml or more (Table 4, Table 5).

According to another object of the present invention, the present invention includes the following steps, and the physiologically active components of quercetin, epigallocatechin, protocatecuic acid and chlorogenic acid are enhanced, and antioxidant activity, anti-diabetes and anti-obesity, etc. It provides a method for producing a vegetable brown sauce with enhanced physiological activity:

I) fermenting the mixed powder consisting of lacquer, brown rice, and soybeans with pine mushroom mycelium, extracting ethanol, and concentrating to 5-10 brix;

Ii) extracting the herbal medicinal material consisting of mixed ginseng, angelica, baeksuoh, cheongoong, licorice and jujube, and concentrated to 5-10 brix;

Iii) fermenting soybeans with Bacillus amyloliquefaciens MGD02 strain to prepare cheonggukjang, drying it, adding water to the dried cheonggukjang, boiling, filtering, and concentrating to 20 to 40 brix to prepare cheonggukjang sauce stock;

Iv) adding the vegetable subsidiary material, the cheonggukjang sauce stock and seasoning prepared in step iii) to the roasted flour (roux), boiling, concentrating and filtering; And

V) adding the extracts prepared in steps i) and ii) to the filtrate prepared in step iv).

Step i) Mixed powder of lacquer Pine mushroom mycelium Manufacture of fermented extract

After fermenting the mixed powder consisting of poison ivy, brown rice and soybeans with pine mushroom mycelium, extract with ethanol and concentrate to 5-10 brix.

The mixed powder, pine mushroom mycelium, fermentation, extraction and concentration are as defined above.

Step ii) Preparation of herbal medicinal extract

Herbal medicinal herbs consisting of mixed ginseng, angelica, baeksuoh, cheonggung, licorice, and jujube are extracted with ethanol and concentrated to 5-10 brix.

Herbal medicine, extraction and concentration are as defined above.

As defined above, if necessary, the extraction of steps i) and ii) may be performed individually or in combination.

Step iii) Cheonggukjang sauce stock Produce

Soybeans are fermented with Bacillus amyloliquefacians MGD02 strain to prepare cheonggukjang, dried, and then added water to the dried cheonggukjang, boiled, filtered, and concentrated to 20-40 brix to prepare cheonggukjang sauce stock.

Beans in the present invention may be used as white tae, seoritae, or rat eyes. After adding 2 to 3 times the volume of water to the beans and soaking for 6 to 24 hours, it is preferable to remove the water, increase the water at 100 to 120°C for 15 to 60 minutes, and cool it to 35 to 45°C. It is not.

Bacillus amyloliquefacience MGD02 according to the present invention is used after being cultured according to a conventional method. Tritic soy broth (TSB; Trytic Soy Broth) As a culture medium cultured for about 12 hours in a liquid medium, it may be seeded, but is not limited thereto. The inoculation amount is preferably made of 2.5~5.0%(v/w).If it exceeds 5.0%(v/w), economic costs are excessive during seed culture, and if it is less than 2.5%(v/w), the fermentation rate of Cheonggukjang is delayed. It can affect the fermentation and quality of Cheonggukjang.

Fermentation is preferably fermented for 2 to 5 days at 35 to 45 ℃.

Drying is preferably performed for 2 to 3 days at 50 to 60° C., but is not limited thereto. By manufacturing dried cheonggukjang, quality change can be prevented by continuous fermentation of cheonggukjang, and odors due to over-fermentation can be prevented, thereby maintaining a certain quality.

The amount of water added to the dried cheonggukjang is preferably 2 to 5 times the volume of the dried cheonggukjang.

Concentration can be done in the usual way.For example, when it first boils over strong heat, lower it to medium heat, boil it for a certain period of time, extract it, filter it on a tray, and boil the extract of cheonggukjang over medium heat for a certain period of time to obtain an appropriate concentration (20 ~ 40 brix). Can be concentrated with.

Step iv) Sauce Base Preparation

To the roasted flour (roux), vegetable subsidiary material, cheonggukjang sauce stock and seasoning prepared in step iii) are added, boiled, concentrated, and filtered to prepare a sauce base.

Roo (roasted flour) can be prepared by pouring olive oil into a pan heated to about 130°C, adding flour, then lowering the heat and stirring until brown.

The vegetable subsidiary material may include one or more from the group consisting of tomato, onion, carrot, celery, garlic, ginger, radish, kiwi, green onion, cabbage, and vegetable broth, but is not limited thereto.

Vegetable broth may be used by adding radish, green onions, mushrooms, kelp, etc. to water and boiled for at least 1 hour, but is not limited thereto.

The seasoning may include one or more from the group consisting of sugar, salt, plum chung, schisandra chinensis, vinegar, wine, bay leaves, and pepper, but is not limited thereto.

Concentration and filtration (of solids) can be carried out in a conventional manner. As an example, if it boils by first heating over strong heat, it can be lowered to medium heat and boiled again for a certain period of time to concentrate to an appropriate concentration, and filtration can be performed using a tray.

Step v) Add extract

The extracts prepared in steps i) and ii) are added to the filtrate prepared in step iv) to complete the brown sauce.

The fermented lacquer mixed powder extract and herbal medicinal herb extract, and the amount of these added are as defined above.

If necessary, after step v), the brown sauce may be sterilized, cooled and/or packaged further.

The brown sauce according to the present invention is 100% vegetable and is a vegetable source capable of replacing meat brown sauce with improved antioxidant activity, improved diabetes and obesity improvement (FIGS. 4A to 6).

Brown sauce according to the present invention includes lacquer mixed powder matsutake mycelium fermentation extract and herbal medicinal extract, physiologically active ingredients such as phenolic acid and flavonol, in particular, quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid. It is strengthened to contain high content.

The fermented extract of lacquer mushroom mixed powder contained in the brown sauce of the present invention does not contain urushiol, so it contains an effective physiologically active ingredient derived from lacquer without causing allergies, etc., and fermented with matsutake mycelium, which is naturally oriented and has excellent palatability. Has an advantage.

Brown sauce according to the present invention has improved antioxidant activity, improved diabetes and improved obesity.

The vegetable brown sauce according to the present invention is 100% vegetable, has enhanced physiologically active ingredients, and has excellent functionality, so it can replace meat sauce in accordance with the modern health-oriented trend, and goes well with beans for vegetarians, and fried or This sauce goes well with smoked chicken or duck meat.

Fig. 1 shows HPLC chromatograms before and after fermentation of fermented matsutake mushroom mycelia of lacquer tree mixed powder (red dotted line indicates urushiol extraction time).
2 shows an example of a manufacturing process diagram of a brown sauce according to the present invention.
3 shows the HPLC chromatogram of the brown source according to the present invention (the red dotted area indicates the urushiol extraction time).
Figure 4 is a graph showing the antioxidant activity of the brown sauce according to the present invention. 4A shows DPPH radical scavenging activity, FIG. 4B shows ABTS radical scavenging activity, FIG. 4C shows hydroxyl radical scavenging activity, and FIG. 4D shows FRAP reducing power.
Figure 5 shows the alpha-glucosidase inhibitory activity of the brown source according to the present invention.
6 shows the pancreas-lipase inhibitory activity of Brown's sauce according to the present invention.

The present invention is explained in more detail by the following examples. These examples are intended to illustrate the invention, and the scope of the invention should not be limited by them.

Preparation Example 1. Fermentation and analysis of lacquer mixed powder pine mushroom mycelium

The fermented product of the lacquer tree mixed powder of the present invention was prepared, and urushiol, which is an allergy-inducing substance, was analyzed before and after fermentation.

<Lacquer mixed powder fermentation>

The lacquer tree was collected from Macheon-myeon, Hamyang-gun and was supplied from the Hamyang-gun Agricultural Technology Center, and the beans were supplied from the South Crop Department of the National Food Service of the Rural Development Administration. .

The pine mushroom mycelium was pre-sold from the forest processing laboratory of Gyeongsang National University, inoculated in 10% white rice liquid medium, and cultured at 25°C for 5 days to prepare a seed.

The lacquer was cut, treated at 100°C for 1 hour, dried at 55°C for 2 to 3 days, and then chopped with a grinder to powder. In addition, beans and brown rice were also pulverized with a grinder to prepare a powder.

After mixing well, 120 g of sumac powder, 40 g of soybean powder, and 40 g of brown rice powder, added purified water to adjust the moisture to about 50%, sterilized at 121°C for 30 minutes to cool, and then the matsutake mycelium spores prepared above were 5 %(v/v) was inoculated, fermented at 25°C for 12 days, dried at 55°C for 2-3 days, and pulverized with a grinder to prepare a powder.

<Sample preparation>

Samples for urushiol analysis were prepared in accordance with the'Urusiol Identification Test' of harmful substances in food in the General Test Methods of the Food Code. Specifically, for comparison with the fermented lacquer mixed powder prepared above, 20 g of each lacquer mixed powder before fermentation was added and 100 ml of 95% ethanol was added, followed by stirring and extraction for at least 3 hours, and then using a Buchner funnel. After suction filtration, the residue and the container were washed with ethanol, and the filtrate was collected and concentrated under reduced pressure in a water bath of 40°C or less. The residue was dissolved in 2 ml of acetonitrile, filtered through a 0.45 μm syringe filter, and prepared as a sample.

<Urusiol analysis>

Urushiol analysis was performed with a high-pressure liquid chromatogram (HPLC, Agilent 1200 series, Agilent Co) by slightly modifying the'Urusiol Identification Test' of harmful substances in food in the General Test Method of the Food Code. The mobile phase solvent was analyzed with 2.0% glassial acetic acid (in water) (solvent A) and acetonitrile (solvent B), and the mobile phase conditions were 10%, 95% for 5, 20, 27, 29 and 30 minutes, respectively, based on solvent B. , 95%, 10% and 10%. 20 µl of the sample was injected and the mobile phase rate was maintained at 1 ml/min at 60° C., and the absorbance of a diode array UV detector (Agilent 1200 series, Agilent Co.) was quantified at 276 nm and is shown in Table 1, and an HPLC chromatogram is shown in FIG. Shown in.

Urushiol content (mg/100 g) Before fermentation 1.52 After fermentation nd All experiments were repeated five times and expressed as an average value.
nd: not detected

As shown in Table 1 and FIG. 1, 1.52 mg/100 g of urushiol, which is an allergen, was detected before fermentation, but it was not detected at all after fermentation, so that the allergen was removed (detoxified) by fermentation.

Preparation Example 2. Preparation of extract of lacquer tree mixed powder matsutake mycelium fermented product and herbal medicine

<Manufacture of herbal medicine powder>

Sanyangsam was prepared by purchasing more than 3 years grown in Seo-myeon, Hamyang-gun, washing 3 times in running water and drying for 2-3 days at 55℃. Ginseng, angelica, baeksuoh, cinnamon, licorice and jujube were dried. It was purchased from Jayeonae Pharmaceutical Co., Ltd. located in Geumseo-myeon, Sancheong-gun.

Wild ginseng, ginseng, angelica, baekshou, cheongoong, licorice and jujube were pulverized into powders, respectively, and mixed ginseng powder was prepared by mixing wild ginseng powder and ginseng powder in a weight ratio of 1:9.

After mixing well mixed ginseng powder 20 g, angelica powder 20 g, baekshou powder 20 g, cheonggung powder 20 g, licorice powder 20 g, and jujube powder 20 g, the lacquer tree mixed powder prepared in Preparation Example 1 matsutake mycelium After 80 g of fermented product powder was added to the mixed powder, 200 ml of 50% ethanol was added to 10 g, diluted 20 times, and extracted by maintaining a 600 rpm speed at 70° C. for 5 hours. Extracted twice under the same conditions, collected, filtered through filter paper (No. 2, Whatman, Tokyo Roshi Kaisha, Ltd., Tokyo, Japan), and concentrated using a reduced pressure concentrator so that the soluble solid content becomes 10 brix. A complex extract of the fermented matsutake mushroom mycelium of lacquer tree mixed powder and herbal medicine was prepared.

Manufacturing example 3. Cheonggukjang Sauce stock Produce

After immersing 1 kg of beans in about 2 times the volume of water for about 12 hours, drain the water for about 2 hours, cook for 30 minutes at 120°C, cool to about 40°C, and put the steamed beans in a fermentation mold, then TSB Inoculated with 2.5% (55 ml) of Bacillus amyloliquefacians MGD02 culture medium cultured for about 12 hours in liquid medium, and fermented at 37°C for 72 hours to prepare Cheonggukjang, which was dried at 50 ~ 60°C for 2 days. Dry cheonggukjang was prepared.

Add 4 L of drinking water to 1 kg of dried cheonggukjang, first heat it over strong heat, and when it boils, lower it to medium heat and boil for 1 hour to extract. Filter it on a tray to get the cheonggukjang extract. The extracting process of cheonggukjang was repeated twice to collect the whole cheonggukjang extract so that it became 4L, then boiled over medium heat and concentrated to about 30 brix to prepare cheonggukjang sauce stock.

Preparation Example 4. Preparation of Brown Sauce

Brown sauce was prepared by using the complex extract of the lacquer tree mixed powder prepared in Preparation Examples 2 and 3, the fermented matsutake mycelium and herbal medicinal herbs, and cheonggukjang sauce stock.

Specifically, olive oil was poured into a pan heated at 130°C, flour was added, and then the heat was lowered and stirred until it became brown while stirring so as not to stick to it to prepare roux. As shown in Table 2, to 400 ml of cheonggukjang sauce stock in Preparation Example 3, 30 g of Lu, carrots, onions, celery, green onions (40 g each), 40 ml of vinegar, 40 ml of Schisandra chinensis, 40 ml of vegetable stock, sugar as a seasoning , Salt, garlic, and ginger were each added 5 g and boiled at 100° C., filtered through a stalk to prepare a sauce, and the extract (soluble solid content: 10 brix) prepared in Preparation Example 2 was each 0 ml (control example) and 40ml (about 5% by weight; Example) was added and boiled at 100° C. for 30 minutes, and then cooled to prepare a brown sauce (FIG. 2).

sample Mixing amount ratio (%) Cheonggukjang Sauce Stock 400 52.2 Lou (fried flour) 30 3.8 Mixed vegetables
(Carrot, onion, celery, green onion: 40 g each) 160 20.8 2x vinegar 40 5.2 Vegetable broth 40 5.2 Sugar and salt (5 g each) 10 1.2 Omija Cheong 40 5.2 Garlic and ginger (5 g each) 10 1.2 extract 40 5.2 Sum 770 100

Test Example 1. Analysis of whether brown sauce contains allergens

The analysis of whether urushiol, which is an allergy-inducing substance to the brown sauce prepared in Preparation Example 4, was contained in the same manner as in Preparation Example 1, and the resulting HPLC chromatogram is shown in FIG.

As shown in Figure 3, in the brown sauce according to the present invention, urushiol, which is an allergen, was not detected.

Test Example 2. Analysis of Bioactive Substances in Brown Sauce

Total phenolics, total flavonoids, phenolic acids, and flavonols, which are physiologically active substances of the brown sauce of Example and Control Example prepared in Preparation Example 4 were analyzed.

<Sample preparation>

The brown sauces of Examples and Controls prepared in Preparation Example 4 were diluted 10 times with tertiary distilled water, respectively, and then filtered through a 0.45 μm filtration filter to prepare a sample.

<Analysis of total phenolics and total flavonoid content>

The total phenolics content was measured by slightly modifying the Folin-Denis method. 0.5 ml of each sample was dispensed into a test tube, 0.5 ml of a 25% Na ₂ CO ₃ solution was added and allowed to stand for 3 minutes. Then, 0.25 ml of a 2 N Folin-Ciocalteu phenol solution was added and mixed, followed by color development at 30° C. for 1 hour. The absorbance of the colored sample was measured using a spectrophotometer (Spectronic 2D) at 750 nm, and a value was calculated from a standard calibration curve prepared using gallic acid, and the results are shown in Table 3.

Total flavonoid content was measured by the Davis method. 0.5 ml of each sample 1.0 ml of diethylene glycol and 0.01 ml of 1 N-NaOH were added and left in a constant temperature water bath at 37° C. for 1 hour. Thereafter, the absorbance was measured at 420 nm using a spectrophotometer (Spectronic 2D), and the value was calculated from a standard calibration curve prepared using a routine, and the results are shown in Table 3.

sample Analysis item Total Phenolic (mg/ml) Total flavonoids (mg/ml) Contrast 0.28 0.26 Example 0.32 0.34 All experiments were carried out in triplicate and expressed as an average value.

As shown in Table 3, the brown sauce according to the present invention (Example) had a total phenolics and total flavonoid content of 0.32 mg/ml (about 14% enhancement) and 0.34 mg/ml (about 30% enhancement), respectively. As a result, it was significantly improved compared to the brown sauce of the control example (no extract was added).

<Phenolic acid and flavonol analysis>

The analysis of phenolic acid and flavonol substances was performed by high-performance liquid chromatography (Agilent 1200 series, Agilent Co, Forest Hill, Vic, Australia). The mobile phase solvents were analyzed with 2.0% glacial acetic acid (in water) (solution A) and 2.0% acetonitrile (in glacial acetic acid) (solution B), and the mobile phase conditions were 10, 15, 20, 25, respectively, based on solvent B. It was held at 15%, 5%, 15%, 5%, 10%, 50%, 50%, 60%, 80% and 90% for 30, 35, 40, 45, 55 and 60 minutes. Each sample was injected with 20 µl, and the mobile phase rate was maintained at 1 ml/min at 30°C, and phenolic acids were quantified at 280 nm absorbance by a diode array UV detector (Agilent 1200 series, Agilent Co.). The bonol (flavonols) material was quantified at an absorbance of 270 nm, and the results are shown in Table 4 and Table 5, respectively.

Content (㎍/ml) Contrast Example Gallic acid 137.34 161.14 Protocatechuic acid 364.60 403.26 Chlorogenic acid 478.26 499.56 p- Hydrobenzoic acid 59.18 82.66 Vanillic acid 25.78 31.66 Ferulic acid 25.49 33.25 Vertaric acid 45.20 65.33 t -Cinnamic acid 1.24 1.33 All experiments were repeated three times and expressed as an average value.

As shown in Table 4, it can be seen that the brown sauce of the Example according to the present invention contains a phenolic acid compound in an increased content compared to the control example, and in particular, protocatechuic acid and chlorgenic acid acid) was contained in high content at 403.26 µg/ml and 499.56 µg/ml, respectively.

Content (㎍/mL) Contrast Example Epigallocatechin 2481.87 2601.56 Catechin 240.29 256.29 Epicatechin 52.06 153.48 Epigallocatechin gallate 28.30 25.57 Rutin 32.91 56.96 Catechin gallate 40.17 88.98 Quercetin 1010.18 1301.23 Naringin 34.75 96.68 Naringenin 81.78 76.22 Formonoetin 8.08 5.24 All experiments were repeated three times and expressed as an average value.

As shown in Table 5, the brown sauce of the examples according to the present invention contained both flavonol compounds in an enhanced content compared to the control example, and in particular epigallocatechin and quercetin were 2601.56 μg/ml, respectively. And 2601.56 μg/ml and contained in a high content.

Test Example 3. Analysis of Antioxidant Activity of Brown Sauce

The antioxidant activity of the brown sources of Examples and Control Examples prepared in Preparation Example 4 was analyzed by measuring DPPH radical scavenging activity, ABTS radical scavenging activity, hydroxyl radical scavenging activity, and FRAP reducing power.

<Sample preparation>

Samples were prepared by diluting the brown sauces of Examples and Controls prepared in Preparation Example 4 1 times (stock solution), 5 times, 10 times, 50 times and 100 times with tertiary distilled water, respectively, and then filtered through a 0.45 μm filtration filter. I did.

<DPPH radical scavenging activity>

DPPH radical scavenging activity was obtained by adding 0.8 ml of DPPH solution (1.5 ^-4 M) to 0.2 ml of each sample diluted 10-fold, 50-fold and 100-fold and filtered, and allowed to stand for 30 minutes after uniformly mixing. It was carried out by measuring the absorbance at nm. The negative control of DPPH radical scavenging activity was performed in the same manner using distilled water instead of the sample, and the difference in absorbance was calculated as a percentage (%) by the following equation, and the result is shown in FIG. 4A.

Radical scavenging activity (%) = [1-(absorbance in negative control ÷ absorbance in experimental section)] ×100

As shown in Fig. 4A, when brown sauce was diluted 10 times, 50 times and 100 times, the examples according to the present invention showed scavenging activities of 77.18%, 44.21% and 11.51%, respectively, and thus the control examples (72.46%, and 100 times). 39.08% and 10.45%) showed enhanced scavenging activity.

<ABTS radical scavenging activity>

ABTS radical scavenging activity was obtained by mixing 5 ml of 7mM ABTS reagent and 5 ml of 140mM K ₂ S ₂ O ₈ (FW 270.3, Sigma 9392) and leaving it in a dark place for 14 to 16 hours to generate cation radicals, which were then mixed with methanol and mixed at 732 nm. The ABTS solution adjusted so that the absorbance value of the control was 0.7±0.02 was used.

0.1 ml of each sample diluted 10 times, 50 times and 100 times prepared above and filtered and 0.9 ml of ABTS solution were mixed, reacted for 3 minutes, and absorbance was measured at 732 nm. The ABTS radical scavenging activity was also negative control using distilled water instead of the sample, and the difference in absorbance was calculated as a percentage (%) by the above equation, and the result is shown in FIG. 4B.

As shown in Figure 4b, the brown sauce of the embodiment according to the present invention all showed enhanced scavenging activity compared to the control example. In particular, when brown sauce was diluted 50 times, the Example according to the present invention exhibited 88.91% of the scavenging activity, which was significantly enhanced compared to the control example (81.54%).

<Hydroxy radical scavenging activity>

The hydroxyl radical scavenging activity is 10mM FeSO _{4 .} 7H ₂ 0-EDTA 0.2 ml, 10 mM 2-deoxyribose 0.2 ml, 10 mM H ₂ O ₂ 0.2 ml, and 1.4 ml of each sample diluted 10 times, 50 times and 100 times prepared above and filtered. After reacting at °C for 4 hours to prepare a mixture, 1 ml of 1% thiobarbital acid (in DW) and 2.8% trichloroacetic acid (in DW) were added to the mixture, followed by color development at 100 °C for 20 minutes and cooling. Absorbance was measured at 520 nm. As a negative control, PBS (1L NaCl 8.76g, NaH ₂ PO ₄ 0.11g, Na ₂ HPO ₄ 0.596g) was used instead of the sample. As for the hydroxyl radical scavenging activity, the difference in absorbance between the addition and no addition of the sample solution was calculated by the above equation, and the results are shown in FIG. 4C.

As shown in Figure 4c, the brown sauce of the embodiment according to the present invention all showed enhanced scavenging activity compared to the control example. In particular, when the brown sauce was diluted 10 times, the Example according to the present invention showed 48.75% of the scavenging activity, which was considerably higher than the control example (41.68%).

<FRAP reducing power>

FRAP reducing power is Acetate buffer (30 mM, pH 3.6), TPTZ reagent (10 mM in 40 mM HCl), and FeCl ₃ The solution (20 mM in DW) was mixed in a ratio of 10:1:1 (v/v/v) to prepare a FRAP measurement reagent, and then pre-reacted for 15 minutes in an incubator at 37° C., and then prepared 10 times, 50 50 µl of each sample diluted twice and 100-fold and filtered, and 950 µl of FRAP reagent were dispensed into a test tube, reacted at 37°C for 15 minutes, and absorbance was measured at 593 nm using a spectrophotometer. In the negative control experiment, an extraction solvent was used instead of the sample, and the results are shown in FIG. 4D.

As shown in Figure 4d, the brown sauce of the embodiment according to the present invention all showed enhanced reducing power compared to the control example. In particular, when the brown sauce was diluted 10 times, the Example according to the present invention exhibited a reducing power of 1.548, indicating a significantly higher reducing power than the control example (1.357).

Test Example 4. Analysis of antidiabetic and antiobesity activity of Brown's sauce

The anti-diabetic activity and anti-obesity activity of the examples and control examples prepared in Preparation Example 4 were determined by measuring the alpha-glucosidase inhibitory activity, an indicator of the anti-diabetic effect, and the pancreatic-lipase inhibitory activity, which is an indicator of the anti-obesity effect. Analyzed.

<Antidiabetic activity>

Alpha-glucosidase inhibitory activity is 50 µl of each sample diluted 1, 5- and 10-fold prepared above and filtered, 50 µl of alpha-glucosidase (0.5 U/ml) enzyme solution, 50 µl of 200 mM sodium phosphate buffer solution (pH 6.8) was mixed and pre-reacted at 37°C for 10 minutes. Thereafter, 100 µl of p- NPG (5 mM) dissolved in a sodium phosphate buffer solution (pH 6.8) was added and reacted again at 37° C. for 10 minutes. Na ₂ CO ₃ in this reaction solution After adding 0.75 ml of (100 mM) to stop the final reaction, the absorbance was measured using a spectrophotometer at 420 nm, and the percentage (%) was calculated by the above equation using distilled water as a negative control, and the results are shown in FIG. Done.

As shown in FIG. 5, Brown sauce (Example) according to the present invention exhibited enhanced alpha-glucosiadase inhibitory activity compared to the control example, and in particular, when diluted 5 times, the Example according to the present invention was 34.87. In %, it showed significantly enhanced inhibitory activity compared to the control example (30.192%).

<Anti-obesity activity>

Pancreatic lipase inhibitory activity is 50 µl of each sample diluted 1, 5- and 10-fold and filtered, 50 µl of pancreatic lipase enzyme solution (1.0 U/mL) and sodium phosphate buffer (100 mM) adjusted to pH 6.8. ) 50 µl was mixed and pre-reacted in a water bath at 37°C for 10 minutes, 100 µl of p- NPB (5 mM) dissolved in sodium phosphate buffer was added, reacted for 10 minutes, and then 0.75 ml of sodium carbonate (100 mM) was added to the enzyme reaction. Was terminated and the absorbance was measured at 420 nm. As the negative control, distilled water was taken instead of the sample, and the difference in absorbance between the addition and the non-addition of the sample solution was calculated as a percentage (%) by the above equation, and the results are shown in FIG.

As shown in Figure 6, Brown sauce according to the present invention (Example) showed significantly enhanced pancreatic lipase inhibitory activity compared to the control example, and Example 59.11%, 5 times when the stock solution (1-fold dilution) was used. At the time of dilution, the inhibitory activity was 35.30%, and at the time of 10-fold dilution, the inhibitory activity was 17.39%, which was significantly higher than that of the control example (38.51%, 13.77%, and 10.66%, respectively).

Test example 5. Brown sauce Physicochemical properties

The physicochemical properties of the brown sauce of Example and Control Example prepared in Preparation Example 4 were analyzed by measuring the pH, acidity, soluble solids, reducing sugar and water-soluble protein contents of.

The pH was measured with a pH meter (MP 200, UK), and the acidity was expressed in% by converting the amount of lactic acid consumed when neutralized to pH 8.2±0.2 using 0.1 N NaOH. It was measured using. As for reducing sugar, 1 ml of DNS reagent was added to 0.1 ml of each analysis sample according to the DNS method, followed by color development at 100°C for 20 minutes, then rapidly cooled, and absorbance was measured at 570 nm using a spectrophotometer and compared with a calibration curve.

The water-soluble protein content was carried out through the biuret method. That is, after dissolving 6.0 g of KNaC ₄ H ₄ O ₆ 4H ₂ O (sodium potassium tartrate) to 1.5 g of CuSO ₄ 5H ₂ O (copper sulfate pentahydrate) in 500 ml of distilled water and adding 300 ml of 10% NaOH, the final volume Biuret reagent was prepared by diagnosing to be 1,000 ml. 1 g of each sample was taken into a test tube, and 4 ml of a biuret reagent was added and mixed thereto, followed by reaction at 37°C for 20 minutes. At this time, the negative control was carried out by taking distilled water, and after 20 minutes reaction, centrifugation for 3 minutes, the supernatant was measured absorbance at 540 nm, and the water-soluble protein was quantified according to the calculation formula calculated by the protein standard curve (bovine serum albumin), and is shown in Table 6. Done.

Analysis item Contrast Example pH 4.71 4.67 Acidity (%, based on lactic acid) 1.75 1.81 Soluble solids (Brix, %) 19.8 20.1 Reducing sugar (mg/ml) 61.7 64.89 Water soluble protein (mg/ml) 15.8 16.1 All experiments were repeated three times and expressed as an average value.

As shown in Table 6, the brown sauce of Example had higher acidity, soluble solid content, reducing sugar content, and water-soluble protein content compared to the control example.

Institute of Agricultural Biotechnology KACC92158P 20161212

Claims (6)

Pine mushroom mycelium fermented extract of mixed powder consisting of poison ivy, brown rice and soybeans; Extracts of herbal medicinal herbs consisting of mixed ginseng of wild ginseng and ginseng, angelica, baeksuoh, cheonggung, licorice and jujube; And Cheonggukjang sauce stock fermented with Bacillus amyloliquefacience MGD02 strain deposited with accession number KACC92158P, and a vegetable brown sauce enriched with physiologically active components of quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid.
The method of claim 1, wherein the fermentation extract is included in an amount of 3 to 6 vol%, the extract of the herbal medicinal material is included in an amount of 3 to 6 vol%, and the cheonggukjang sauce stock is included in an amount of 50 to 60 vol%. A vegetable brown sauce enriched with the physiologically active components of quercetin, epigallocatechin, protocatecuic acid and chlorogenic acid.
The mixed powder of claim 1, wherein the fermentation extract is composed of lacquer tree powder 60-80% (w/w), brown rice powder 10-20% (w/w), and soybean powder 10-20% (w/w). After adding water to 50~60% moisture, swelling for 1~2 hours, sterilizing at 100~120℃ for 30~60 hours, inoculating pine mushroom mycelium at 3~6% (v/w) to ferment semi-solid After that, the physiologically active components of quercetin, epigallocatechin, protocatecuic acid and chlorogenic acid, characterized in that it is prepared by extracting with 40-70% ethanol and filtering to adjust the soluble solid content to 5-10 brix, are enhanced Vegetable brown sauce.
The method of claim 1, wherein the brown source is fortified with quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid, respectively, at 1,300 μg/ml or more, 2,600 μg/ml or more, 400 μg/ml or more, and 490 μg/ml or more. A vegetable brown sauce enriched with physiologically active components of quercetin, epigallocatechin, protocatecuic acid and chlorogenic acid, characterized in that it is.
The vegetable brown sauce according to claim 1, wherein the physiologically active components of quercetin, epigallocatechin, protocatecuic acid, and chlorogenic acid are enhanced, which have enhanced antioxidant activity, antidiabetic activity and antiobesity activity.
A method for preparing a vegetable brown sauce enriched with physiologically active components of quercetin, epigallocatechin, protocatecuic acid and chlorogenic acid, the method comprising:
I) fermenting the mixed powder consisting of lacquer, brown rice, and soybeans with pine mushroom mycelium, extracting ethanol, and concentrating to 5-10 brix;
Ii) extracting the herbal medicinal material consisting of mixed ginseng, angelica, baeksuoh, cheongoong, licorice and jujube, and concentrated to 5-10 brix;
Iii) After fermenting soybeans with Bacillus amyloliquefaciens MGD02 strain deposited under the accession number KACC92158P to prepare cheonggukjang, dry it, add water to the dried cheonggukjang, boil it, filter it, and concentrate it to 20~40 brix to prepare cheonggukjang sauce stock. Manufacturing steps;
Iv) adding the vegetable subsidiary material, the cheonggukjang sauce stock and seasoning prepared in step iii) to the roasted flour (roux), boiling, concentrating and filtering; And
V) A method comprising the step of adding the extracts prepared in steps i) and ii) to the filtrate prepared in step iv).

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