KR102199387B1 - Teriyaki Sauce having high biological active materials and preparation method thereof - Google Patents

Abstract

In the present invention, 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 teriyaki sauce containing a high content of physiologically active components of quercetin, epigallocatechin, and chlorogenic acid, including cheonggukjang sauce stock, and a method for producing the same are disclosed.
Teriyaki sauce according to the present invention has improved antioxidant activity, improved diabetes and improved obesity.

Description

Teriyaki Sauce having high biological active materials and preparation method thereof}

The present invention relates to a teriyaki sauce with improved physiologically active ingredients and a method for producing 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 teriyaki sauce containing a high content of physiologically active components of quercetin, epigallocatechin, and chlorogenic acid, including cheonggukjang sauce stock, 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.

Teriyaki is the name of a Japanese dish in which seafood is coated with soy sauce and grilled. This Japanese-style soy sauce is called teriyaki sauce. Teriyaki sauce (sauce) is a soy sauce-based sauce that has a thick consistency, sweetness, and saltiness, and is widely applied to meat dishes as well as fish and shellfish due to its characteristic salty and sweet taste. Boil it with soy sauce, liquor, kelp, and sugar.

Technologies for improving palatability or adding functionality to teriyaki sauce are being developed. For example, teriyaki sauce (Registration No. 10-1919694) with improved palatability by adding charcoal fire, teriyaki sauce having lipolytic enzyme inhibitory properties by containing the ethyl acetate layer fraction of Madi grass extract as an active ingredient (Publication No. 10-2015- 0064724) and the like are known, but development of teriyaki sauce excellent in palatability or functionality using new materials is still 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.

Chlorogenic acid, a type of phenolic acid, is known for its antibacterial and anti-inflammatory properties.

Therefore, the development of teriyaki sauce containing the flavonol and phenolic acid components in a high content 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 Teriyaki sauce containing extracts of herbal medicinal herbs composed of jujube together with cheonggukjang sauce stock contains high content of physiologically active ingredients such as quercetin, epigallocatechin and chlorogenic acid, and has excellent functionality (enhanced physiological activity). After confirming that it had, 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 cheonggukjang sauce stock, to provide a teriyaki sauce containing quercetin, epigallocatechin, and chlorogenic acid, and having an enhanced functionality (physiological activity) and a method for producing 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 components of quercetin, epigallocatechin and chlorogenic acid are enhanced, and teriyaki sauce having excellent functionality is provided.

Lacquer mixed powder Pine mushroom mycelium Fermented extract

The teriyaki sauce of the present invention includes a fermented extract of matsutake mycelia 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 the amount of lacquer powder is less than 60%, the physiological activity may not be sufficient. If the amount of lacquer powder is less than 60%, the amount of brown rice 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 about 50-60% moisture to the mixed powder, swelling it for about 1 to 2 hours, sterilizing at 100 to 120°C for 30 to 60 hours, and then adding 3 to 6% (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 tree mixed powder according to the present invention prepared as described above was detoxified by removing the allergen, Woori Siul (Table 1, Fig. 1), 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 and the like.

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

In the teriyaki sauce of the present invention, the fermented extract of lacquer mixed powder is added in an amount of 3 to 6% by volume when the total weight of the teriyaki sauce is 100% by volume. If it 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 teriyaki sauce of the present invention includes an extract of herbal medicinal herbs consisting of mixed ginseng of wild ginseng and ginseng, angelica, baekshou, 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.

Herbal medicine extract from the teriyaki sauce of the present invention is added in an amount of 3 to 6% by volume when the total weight of the teriyaki 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

The teriyaki 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, as the cheonggukjang sauce stock, preferably, cheonggukjang sauce stock fermented with Bacillus amyloliquefacience MGD02 strain is used.

The present inventors isolated and identified'Bacillus amyloliquefacience 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.

The source stock of Cheonggukjang is dried Cheonggukjang fermented with Bacillus amyloliquefacience MGD02 strain to prepare dried Cheonggukjang, then added 2 to 5 volume times of water to the dried Cheonggukjang, boiled, extracted, filtered, and then made 20 to 40 brix. It is prepared by concentration.

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

In the teriyaki 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 teriyaki 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.

Teriyaki sauce according to the present invention including the above extracts and cheonggukjeong sauce stock contains physiologically active components of quercetin, epigallocatechin, and chlorogenic acid, respectively, of 1,450 µg/ml or more, 2,480 µg/ml or more, and 450 It was enhanced to more than μg/ml (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 and chlorogenic acid are enhanced, and physiological activities such as antioxidant activity, anti-diabetes and anti-obesity are enhanced. Provides a method of making an enhanced teriyaki sauce:

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 cheonggukjang sauce stock prepared in step iii) to a mixture of soy sauce, vinegar, sugar, and vegetable stock, 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, if it boils at first by heating it 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 again for a certain period of time over medium heat to obtain an appropriate concentration. Can be concentrated.

Step iv) Sauce Base Preparation

To a mixture of soy sauce, sugar, vinegar and vegetable stock, the cheonggukjang sauce stock prepared in step iii) is added to boiled, concentrated, and filtered to prepare a sauce base.

Vegetable stock uses radish, green onions, mushrooms, kelp, etc., and boiled for at least 1 hour.

In the mixture, soy sauce, sugar, vinegar, and vegetable stock may be mixed in a volume ratio of 2 to 1: 2 to 1: 1 to 0.5: 2 to 1.

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 teriyaki 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 teriyaki sauce may be sterilized, cooled and/or packaged further.

Teriyaki source according to the present invention is a vegetable source with improved antioxidant activity, improved diabetes and improved obesity (FIGS. 4A to 6 ).

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

The fermented extract of mixed powder of lacquer matsutake mycelium contained in the teriyaki 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.

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

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

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).
Figure 2 shows an example of a manufacturing process diagram of the teriyaki sauce according to the present invention.
3 shows the HPLC chromatogram of the teriyaki sauce according to the present invention (the red dotted area indicates the urushiol extraction time).
Figure 4 is a graph showing the antioxidant activity of teriyaki 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 teriyaki source according to the present invention.
Figure 6 shows the pancreatic-lipase inhibitory activity of teriyaki 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.

Manufacturing example 1. Mixed powder of lacquer Pine mushroom mycelium Fermentation and analysis

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 bean was supplied from the South Crop Department of the National Food Service of the Rural Development Administration, and brown rice was purchased on the market. .

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 extracting 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.

Manufacturing example 2. Mixed powder of lacquer Pine mushroom mycelium Fermented products Manufacture of extracts of 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, baeksuoh, 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 ratio of 1: 9 (w/w).

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 To the mixed powder obtained by mixing 80 g of the fermented product powder, 200 ml of 50% ethanol was added to 10 g, diluted 20 times, and extracted by maintaining a speed of 600 rpm for 5 hours at 70°C. Extracted twice under the same conditions, collected, filtered with filter paper (No. 2, Whatman, Tokyo Roshi Kaisha, Ltd., Tokyo, Japan), and concentrated using a vacuum 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 soaking 1 kg of beans in about twice the amount of water for about 12 hours, drain the water for about 2 hours, cook for 30 minutes at 120℃, cool to about 40℃, and put the steamed beans in the fermentation mold, and then liquid TSB Inoculated with 2.5% (55 ml) of Bacillus amyloliquefacience MGD02 culture medium cultured for about 12 hours in a medium, and fermented at 37°C for 72 hours to prepare Cheonggukjang, which was dried at 50 to 60°C for 2 days and dried. 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.Add 2L of drinking water to the filtered cheonggukjang bean and boil it for 30 minutes. 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.

Manufacturing example 4. Teriyaki Sauce manufacturing

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

Vegetable stock was prepared by adding 800 g of radish, 150 g of green onions, 30 g of dried shiitake mushrooms, and 60 g of dried seaweed to 6,000 mL of water, boiled at 100° C. for 1 hour, cooled, and filtered through the solids.

Specifically, as shown in Table 2, in a mixture of 80 ml of soy sauce, 80 ml of brown sugar, 120 ml of vegetable stock, and 40 ml of vinegar, 400 ml of the cheonggukjang sauce stock in Preparation Example 3 was mixed to reach 200 ml over medium heat. After concentration, sieve through a sieve, to 190 ml of the concentrate, 0 ml (control example) and 10 ml (about 5 vol%; Example) of the extract (soluble solid content: 10 brix) prepared in Preparation Example 2 were added, respectively, and teriyaki sauce Was prepared (Fig. 2).

sample Mixing amount ratio (%) Cheonggukjang Sauce Stock 400 52.6 Vegetable broth 120 15.8 2x vinegar 40 5.3 Brown sugar 80 10.5 Soy sauce 80 10.5 extract 10 (converted value 40) 5.3 Sum 100

Test example One. Teriyaki Analysis of whether the source contains allergens

The analysis of whether urushiol, which is an allergy-inducing substance, to the teriyaki sauce of the Example prepared in Preparation Example 4 was performed in the same manner as in Preparation Example 1, and the resulting HPLC chromatogram is shown in FIG. 3.

As shown in Figure 3, the teriyaki sauce according to the present invention did not detect the allergen urushiol.

Test example 2. Teriyaki Analysis of bioactive substances in source

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

<Sample preparation>

The teriyaki sauces of Examples and Control Examples 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. To 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 0.32 0.17 Example 0.35 0.19 All experiments were carried out in triplicate and expressed as an average value.

As shown in Table 3, teriyaki sauce (Example) according to the present invention has a total phenolics and total flavonoid content of 0.35 mg/ml (approximately 9.3% enhancement) and 0.19 mg/ml (approximately 12% enhancement), respectively. As a result, it was significantly improved compared to the teriyaki sauce (extract was not added) of the control example.

<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 (μg/ml) Contrast Example Gallic acid 131.78 133.70 Chlorogenic acid 342.20 453.14 Vanillic acid 25.20 27.46 p -Coumaric acid nd 36.08 Vertaric acid 14.96 15.97 t -Cinnamic acid 3.01 3.05 All experiments were repeated three times and expressed as an average value.

As shown in Table 4, the teriyaki sauce of the examples according to the present invention had an increased content of a phenolic acid compound, and in particular, chlorgenic acid contained a high content of 453.14 µg/ml.

Content (μg/mL) Contrast Example Epigallocatechin 2317.72 2483.87 Catechin 258.97 272.99 Epicatechin 106.77 117.28 Epigallocatechin gallate 45.79 50.82 Rutin 107.85 118.04 Quercetin 1188.64 1457.62 Naringin 88.56 104.76 Naringenin 36.34 41.21 All experiments were repeated three times and expressed as an average value.

As shown in Table 5, the teriyaki sauce of the examples according to the present invention had an increased content of flavonol compounds, and in particular epigallocatechin and quercetin were 2483.87 μg/ml and 1457.62 μg/ml, respectively. It contained in a high content.

Test example 3. Teriyaki Source's antioxidant activity assay

The antioxidant activity of the teriyaki source 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>

The teriyaki sauce of Example and Control Example prepared in Preparation Example 4 was diluted 1, 5, 10, 50, and 100 times with 3 distilled water, respectively, and then filtered through a 0.45 μm filtration filter to prepare a sample. 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 Figure 4a, when the teriyaki sauce was diluted 10 times, 50 times and 100 times, the examples according to the present invention exhibited scavenging activities of 80.45%, 46.1% and 11.81%, respectively, and thus the control examples (75.97%, and 100 times). 41.9% and 11%) 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 teriyaki sauce of the embodiment according to the present invention all showed enhanced scavenging activity compared to the control example. When teriyaki sauce was diluted 10-fold, 50-fold, and 100-fold, the examples according to the present invention exhibited 99.76%, 98.87% and 44.53% of scavenging activity, respectively, compared to the control examples (99.53%, 95.99% and 41.33%). Showed enhanced scavenging activity.

<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 teriyaki sauce of the embodiment according to the present invention all showed enhanced scavenging activity compared to the control example. In particular, when the teriyaki sauce was diluted 50 times, the Example according to the present invention exhibited a scavenging activity of 19.29%, showing a significantly higher scavenging activity compared to the control example (13.13%).

<FRAP reducing power>

FRAP reducing power is 10:1:1 (v/v/v) in Acetate buffer (30 mM, pH 3.6), TPTZ reagent (10 mM in 40 mM HCl), and FeCl ₃ solution (20 mM in DW). After mixing to prepare a FRAP measuring reagent, pre-reacting for 15 minutes in a 37°C incubator, 50 µl of each sample diluted 10 times, 50 times and 100 times and filtered and 950 µl of FRAP reagent prepared above were dispensed into a test tube. The reaction was performed 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 teriyaki sauce of the embodiment according to the present invention all showed enhanced reducing power compared to the control example. In particular, when the teriyaki sauce was diluted 10 times, the example according to the present invention exhibited a reducing power of 1.535, which was considerably higher than that of the control example (1.384).

Test example 4. Teriyaki Source of Antidiabetic And Anti-obesity Activity assay

The anti-diabetic activity and anti-obesity activity of the teriyaki sauce of Examples and Control Examples prepared in Preparation Example 4 were determined through measurement of alpha-glucosidase inhibitory activity, an indicator of anti-diabetic effect, and pancreatic-lipase inhibitory activity, an indicator of 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 Figure 5, the teriyaki sauce (Example) according to the present invention exhibited an enhanced alpha-glucosiadase inhibitory activity compared to the control example, particularly when diluted 5 times, the Example according to the present invention It showed significantly enhanced inhibitory activity compared to the control example (17.54%) at 20.5%.

<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 Fig. 6, the teriyaki sauce (Example) according to the present invention exhibited significantly enhanced pancreatic lipase inhibitory activity compared to the control example, and in Example, 38.66%, 5 times when diluted with a stock solution (1 times) At the time of dilution, the inhibitory activity was 23.21%, and at the time of 10-fold dilution, 13% was shown, and significantly higher inhibitory activity was shown compared to the control examples (29.14%, 20.24%, 8.14%, respectively).

Test example 5. Teriyaki Physicochemical properties of the sauce

The physicochemical properties of the teriyaki sauce of Examples and Control Examples 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 5.02 5.0 Acidity (%, based on lactic acid) 0.53 0.55 Soluble solids (Brix, %) 25.1 25.8 Reducing sugar (mg/ml) 46.78 49.65 Water soluble protein (mg/ml) 13.6 14.4 All experiments were repeated three times and expressed as an average value.

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

Institute of Agricultural Biotechnology KACC92158P 20161212

Claims (6)

Teriyaki sauce with enhanced physiologically active components of quercetin, epigallocatechin and chlorogenic acid. The teriyaki sauce
Mixed powder consisting of sumac powder 60-80% (w/w), brown rice powder 10-20% (w/w) and soybean powder 10-20% (w/w) is fermented with pine mushroom mycelium and extracted with ethanol. Pine mushroom mycelium prepared by concentrating with 5-10 brix 3-6% by volume of fermented extract;
Mixed ginseng of wild and ginseng 15~25%(w/w), Angelica 15~25%(w/w), Baeksuo 15~25%(w/w), Cheongung 15~25%(w/w), 3-6% by volume of extracts of herbal medicinal herbs prepared by ethanol extraction of oriental medicinal herbs consisting of 15-25% (w/w) licorice and 15-25% (w/w) jujubes and concentrating to 5-10 brix; And
Cheonggukjang sauce stock 50 prepared by fermenting soybeans with Bacillus amyloliquefaciens MGD02 strain deposited under the accession number KACC92158P to prepare and dry Cheonggukjang, add water to the dried Cheonggukjang, boil, filter, and concentrate to 20-40 brix. Contains ~60% by volume,
Teriyaki sauce, characterized in that the quercetin, epigallocatechin, and chlorogenic acid are fortified to 1,450 µg/ml or more, 2,480 µg/ml or more, and 450 µg/ml or more, respectively.
delete 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 matsutake mycelium at 3~6% (v/w) to ferment semi-solid Teriyaki sauce enriched with physiologically active components of quercetin, epigallocatechin 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.
delete delete A method for preparing teriyaki sauce enriched with physiologically active components of quercetin, epigallocatechin and chlorogenic acid according to claim 1, wherein the method comprises:
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 make cheonggukjang sauce stock Manufacturing steps;
Iv) adding the cheonggukjang sauce stock prepared in step iii) to the mixture of soy sauce, vinegar, sugar and vegetable stock, boiling, concentrating and filtering; And
V) adding the extracts prepared in steps ii) and ii) to the filtrate prepared in step iv),
Wherein the quercetin, epigallocatechin, and chlorogenic acid are fortified to 1,450 μg/ml or more, 2,480 μg/ml or more, and 450 μg/ml or more, respectively.

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