KR20110001359A - A method for preparing functional concentrate of black galic - Google Patents

Abstract

PURPOSE: A producing method of a functional aged garlic concentrate is provided to enable a user to easily eat garlic by using an aged garlic extract, a Japanese apricot extract, and an agaricus blazei mycelium extract. CONSTITUTION: A producing method of a functional aged garlic concentrate comprises the following steps: ferment-aging garlic into aged black garlic; extracting the aged garlic with a solvent for 1~48hours at room temperature, and condensing at 30~100deg C to obtain an aged garlic extract; mixing Japanese apricot with sugar before fermenting for 1~6months at 10~100deg C, and obtaining a Japanese apricot extract by filtering the mixture; extracting an agaricus blazei mycelium with the solvent for 1~48hours at room temperature, and filtering to obtain an agaricus blazei mycelium extract; and mixing the aged garlic extract, the Japanese apricot extract, and the agaricus blazei mycelium extract before sterilizing, dispensing, and sealing.

Description

A method for preparing functional concentrate of black galic

The present invention relates to a method for producing a functional black garlic concentrate having excellent nutrition, flavor, palatability, and anticancer effect using plum extract and agaricus mushroom mycelium extract as main ingredients.

[Document 1] Seo Hwa-jung, Korean Journal of Food Science and Nutrition, 29 (3) , pp.485-492, 2000

[Reference 2] Cho Hyun-joo et al., Korean Journal of Food Science and Nutrition, 31 (1) , pp.98-103, 2002

[3] Huh Geun et al., Korean Journal of Pharmacy, 45 (3) , pp.258-268, 2001

[School 4] Kim, Sun-Min, Food Industry and Nutrition, 4 (3) , pp.55-58, 1999

[Reference 5] Park, Moo-hyun et al., Korean Journal of Food Science and Nutrition, 29 (1) , pp.128-133, 2000

[Reference 6] Young Ho Baek, Korean Journal of Nutrition and Nutrition, 24 (6) , pp.970-977, 1995

[Document 7] Park Jong-hee, Encyclopedia of Herbal Medicine (Ha), Book Publishing Company, Shin-il, p.574-575, 2002

Carmichael et al., Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemisensitivity testing. Cancer Research , 47 , pp. 944-946, 1987

Fruit beverage consumption in Korea was 13.1% based on soft drink production in 1987, and grew to 23.7% in 1991, accounting for one third of 1.5 trillion won in soft drink production. (Ministry of Health and Welfare, 1991). This trend will increase as consumers' preference for natural juices increases. In fact, the market share of natural juices was only 62.9 billion won in 1995, but grew rapidly to 200 billion won in 1996.

Fruits and vegetables are considered as health foods because they contain a large amount of vitamins, minerals, fiber, enzymes and other pharmacological ingredients. Fruit and vegetable juices have the advantage of reducing intestinal burden and ingesting large amounts of active ingredients. have. In particular, the effects of mixing several vegetables or fruit juices are greater than consuming only one type of juice solution alone. Therefore, the processing process for producing high quality juices using domestically produced fruits and vegetables is more effective. Optimization is an urgent research task, and moreover, it is urgently required to provide functionality or quality in keeping with the increasing and diversifying preferences of consumers.

In addition, increasing the productivity of agriculture and creating value-added and consumed domestic fruits and vegetables by diversifying relatively low-use agricultural products will eventually benefit both farmers, businesses and consumers.

Garlic is a perennial bulb plant belonging to the genus Liliumaceae Allium sativum for peiinense Makino (Alium scorodoprassum L.). With the development of China, it is spread to Arabia, Afghanistan, India, and China, and is now cultivated all over the world. Garlic is mainly composed of 56 ~ 68% water, 26 ~ 30% carbohydrate, 2.3% organosulfur compound, 2% protein (mainly allinase), 1.5% fiber, etc. It is known to be due to allicin (allicin), one of the organic sulfur compounds of the allicin is a compound having a very low solubility in water, exists in the garlic (alliin) in garlic. The structure of garlic is a protein called allin, which does not have a bad smell on its own. However, when an external change such as a wound, a knife or a grind on the surface of the garlic occurs, it has a unique smell. The alinase enzyme in the body changes into allicin, which has a strong smell and pungent taste due to contact with oxygen. Garlic lowers blood pressure and lowers lipids (Seohwajung, Korean Journal of Food and Nutrition, 29 (3) , pp.485-492, 2000; Cho, Hyun-joo et al., Korean Journal of Food and Nutrition Science, 31 (1) , pp.98- 103, 2002), antioxidant activity (Hu Geun et al., Korean Journal of Pharmacy, 45 (3) , pp.258-268, 2001; Kim Sun-min, Food Industry and Nutrition, 4 (3) , pp.55-58, 1999), immune (bakmuhyeon et al., Journal of the Korea Food and nutrition, 29 (1), pp.128-133, 2000), anti-fatigue effects (Journal baekyoungho, Korea Food and nutrition, 24 (6), pp.970-977, 1995).

Plum is a fruit of plum tree, round shape, ripen in green from the end of May to the middle of June. It is originated from China, and has been used as a health supplement or medicine since 3000 years ago. It is assumed to be used as. It can be seen that the Chinese medicine book, Xin Nong-Kwon, about 2000 years ago, was used as a medicine.In addition, the effect is recorded in Dongbobom and Herbal Wood, and especially in the Chinese Herbal Wood. It is. Plum fruit is about 80% of the fruit, of which about 85% is water and the sugar is about 10%. It contains a large amount of organic acids, minerals such as calcium and potassium, vitamins such as carotene and the like. Among them, the main component of organic acid is citric acid.In addition, it contains citric acid, malic acid, succinic acid, etc.In addition, organic acid activates gastrointestinal action and stimulates appetite, and citric acid promotes metabolism of sugar and releases fatigue. give. Plum is an alkaline food, good for fatigue recovery and constitution improvement (Park Jong-hee, Encyclopedia of Herbal Medicine, Hae, Shinil Trading Co., p.574-575, 2002).

Agaricus blazei is a genus of fungi, fungus, coliform , agaric, and fungi. It is native to Brazil and grows in a mountainous area called Piedade in Brazil. Dr. WJ Synden of Pennsylvania State University and ED Lambert, Ph.D., of the Lambert Institute, said the mushrooms were known to have caused mushrooms called Agaricus, which causes local people to suffer from disease and other diseases. In Brazil, it is called Royal Agaricus or Mushroom of the Sun. In Korea, it is called White Mushroom or Spirit Mushroom. In the same genus, there are mushrooms, and in the same throat, mushrooms, enoki mushrooms and oyster mushrooms. The effect is known.

However, none of the above documents teaches or describes any preparation method for producing functional black garlic concentrate using the black garlic, plum and agaricus mushroom mycelium of the present invention.

Therefore, the present inventors are studying a method to increase the palatability and flavor of garlic, which is not easy to ingest due to its specific smell, and prepares black garlic concentrate by using a black garlic extract as a main ingredient, a plum extract and agaricus mushroom mycelium extract. Not only excellent flavor and palatability, but also shows an excellent anti-cancer effect of inhibiting the proliferation of cancer cells, the production method of functional black garlic concentrate was completed in the present invention.

In accordance with the above object, the present invention provides a manufacturing method for producing a functional black garlic concentrate having high nutrition, flavor, palatability and anticancer effect using black garlic.

Specifically, the present invention, the black garlic concentrate is the first step of fermenting garlic with black garlic fermentation; Removing the skin of the black garlic prepared in step 1 and hot water extraction, cold immersion of a solvent selected from 1 to 30 times the volume of the sample weight water, lower alcohol of 1 to 4 carbon atoms or a mixed solvent thereof at room temperature for 1 to 48 hours A second step of extracting by extraction method such as extraction, ultrasonic extraction, reflux cooling extraction, heating extraction, and concentrating under reduced pressure at a temperature of 30 to 100 ° C. to obtain black garlic extract; Plums and sugar are mixed at a ratio of 1-20: 1-20 (w / w / w), preferably 1-10: 1-10 (w / w / w), and 1-6 at a temperature of 10-100 ° C. Fermentation for months, followed by filtration to obtain plum extract; Soybean digested extract was added to the incubator, sterilized, and then inoculated with 50 to 1000 milliliters of mushroom spawn to incubate for 24 to 144 hours to obtain mycelium. Or a fourth step of extracting and filtering the solvent selected from these mixed solvents by extraction methods such as hot water extraction, cold sediment extraction, ultrasonic extraction, reflux cooling extraction, and heat extraction at room temperature for 1 to 48 hours to obtain mushroom mycelium extract; The black garlic extract, plum extract and mushroom mycelium extract prepared above are mixed at a mixing ratio of 50 to 90:10 to 40: 1 to 10 (w / w / w), and then sterilized, and a predetermined amount, preferably 10 to 10 per ampule. After dispensing 40 mL, the functional black garlic concentrate of the present invention can be prepared from the manufacturing process, which comprises a fifth step of sealing to obtain a black garlic concentrate.

The black garlic concentrate as defined herein is characterized in that the concentrate is a mixture of black garlic extract, plum extract and mushroom mycelium extract.

Mushrooms as defined herein are mushroom mushrooms, matsutake mushrooms, shiitake mushrooms, agaricus, eggplant mushrooms, roe deer mushrooms or chaga mushrooms, and the like, and preferably include mushrooms, matsutake mushrooms or agaricus mushrooms.

Hereinafter, the present invention will be described in detail.

The black garlic extract of the present invention can be obtained as follows.

For example, (a) in order to prepare black garlic extract, garlic, preferably domestic Namhae village, domestic Uiseong garlic or domestic Muan garlic, more preferably about 10 to 120 ℃ grown garlic grown in Namhae-gun, Gyeongsangnam-do, Korea, A first step of first aging with hot air, preferably at about 40 to 90 ° C. for about 150 to 500 hours, preferably about 280 to 320 hours; A second step of naturally drying garlic aged in the step for 10 to 60 hours, preferably 38 to 42 hours; Third step comprising the second ripening step of ripening the dried garlic for 10 to 100 hours, preferably 30 to 50 hours with hot air at a temperature of 10 to 50 ℃, preferably 20 to 30 ℃ After grinding the black garlic obtained in step 3, a solvent selected from 1 to 30 times the weight of the sample, preferably 2 to 15 times the volume of water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water Extraction methods such as hot water extraction, cold sediment extraction, ultrasonic extraction, reflux cooling extraction, heat extraction, and the like, selected from ethanol, a solvent selected from ethanol, more preferably water, for 1 to 48 hours, preferably 3 to 12 hours. After the extraction by hot water extraction method, the black is concentrated through a fourth step of manufacturing the black garlic extract by concentrating under reduced pressure at a temperature of 30 to 100 ° C, preferably 50 to 100 ° C. You can prepare a garlic extract.

(b) To prepare a plum extract, the plums are washed with water and the plums and sugars are 1-20: 1-20 (w / w / w), more preferably 1-10: 1-1-10 (w / w / w), even more preferably, 1 to 6 months, preferably 2 at a temperature of 10 to 100 ° C., preferably 15 to 70 ° C., mixed with each other at a mixing ratio of about 1: 1 (w / w / w) After fermentation for 5 to 5 months, the plum extract may be prepared through a process of preparing the filtered plum extract.

In addition, (c) in order to prepare a mushroom extract, the first step of putting the soybean decomposition extract in the incubator, sterilization; After cooling the sterilized solution prepared in step 1, mushroom seed, preferably Agaricus blazei seed, is inoculated at 50 to 1000 milliliters, preferably 200 to 700 milliliters, for 24 to 144 hours, preferably 48 A second step of incubation for 120 hours; A third step of obtaining the mycelia grown by growing the mycelia cultured in step 2; A fourth step of obtaining the mushroom mycelium powder by pulverizing and drying the mycelium; A solvent selected from 1 to 30 times the weight of the powder sample, preferably 5 to 15 times the volume of water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water or a solvent selected from ethanol, more preferably Preferably, the extract available in water is extracted for 1 to 48 hours, preferably 5 to 12 hours, at room temperature for extraction, such as hot water extraction, cold extraction, ultrasonic extraction, reflux cooling extraction, heating extraction, and preferably hot water extraction. A fifth step; After filtering the extract of step 5 and concentrated under reduced pressure at a temperature of 30 to 100 ℃, preferably 50 to 100 ℃ can be prepared mushroom mycelium extract through a process for producing agaricus mushroom mycelium extract.

Each of the composition components obtained in the above production method, that is, (a) black garlic extract, (b) plum extract and (c) mushroom mycelium extract is a mutually constant mixing ratio, preferably (a) black garlic extract, (b) plum The dry weight mixing ratio of the extract and (c) the mushroom mycelium extract is 50 to 90: 10 to 40: 1 to 10 (w / w / w), more preferably 60 to 80: 15 to 30: 3 to 8 ( w / w / w), more preferably, in a mixing ratio of about 70: 25: 5 (w / w / w), mixed with each other and sterilized to obtain a black garlic concentrate; After dispensing the black garlic concentrate sterilized in step 1 in a predetermined amount, preferably 10 to 40 mL per ampoule, more preferably 15 to 30 mL, the black garlic concentrate of the present invention may be obtained through the manufacturing process of the second step of sealing. Can be.

As described above, the black garlic concentrate of the present invention is prepared by using the black garlic extract as a main ingredient, plum extract and Agaricus mushroom mycelium extract, not only excellent in high nutrition, flavor and palatability, but also excellent anti-cancer effect of inhibiting cancer cell proliferation. Since it can be prepared as a functional black garlic concentrate of excellent quality.

Below, The invention is illustrated in detail by the following examples and experimental examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Examples, Reference Examples and Experimental Examples.

Example 1 Preparation of Black Garlic Extract

Domestic fresh garlic grown in Namhae-gun, Gyeongsangnam-do is purchased from Namhae Agricultural Cooperatives, and 500g of fresh garlic is aged at 40-90 ℃ for about 280-320 hours by hot air, and the mature village is naturally dried for about 38-42 hours, and then 20-30 ℃. The hot air was aged for about 30 to 50 hours to obtain 347.5 g of black garlic. After removing the bark of the obtained black garlic, three times the purified water was added to the weight of black garlic and extracted by hydrothermal extraction for 5 hours at a temperature of 100 ℃ using an extraction concentrator (Sama Engineering, Korea). The black garlic hot water extract was concentrated under reduced pressure with a vacuum rotary concentrator (Tokyo Rikakikai Co. Ltd., EYELA) at a temperature of 70 ℃ to obtain a black garlic concentrate of 30 ° Brix.

Example 2 Preparation of Plum Extract

1kg of plum and 1kg of white sugar purchased from Changnyeong-gun, Gyeongsangnam-do, Korea are mixed at a ratio of 1: 1 (w / w), fermented at 20-25 ℃ for about 3 months, and then filtered and the plum extract (36.6 ° Brix) Obtained.

Example 3 Preparation of Agaricus Mycelium Extract

1 liter of soybean digestion extract (Brix: 8) was put into an 18 liter incubator and sterilized according to a conventional method. After the sterile liquid was sufficiently cooled, Agaricus blazei species purchased from domestic HK Biotech (Jinju, Gyeongsangnam-do) was inoculated with 200 to 400 milliliters and incubated for 96 hours. The mycelium was grown to about 75% of the culture solution, and then transferred to the extraction container, water was added at a ratio of 1:10 (w / v), mixed, and extracted at 100 ° C. for 2 hours. In order to remove solids from the extracted solution, the resultant was filtered through a filter to obtain Agaricus mycelium extract (36.6 ° Brix).

Example 4. Preparation of Black Garlic Concentrate

The black garlic, plum and Agaricus mushroom mycelium extracts obtained in Examples 1 to 3 were diluted with purified water and used to adjust to 30 ° Brix. Black garlic extract: Agaricus mushroom mycelium extract was mixed at a ratio of 70: 25: 5 (v / v), sterilized by autoclaving at 121 ° C. for about 10 minutes, and put in a 20 ml glass bottle with a dispenser (Ceramus, Germany) and covered with a rubber stopper. Seal with aluminum seal.

Reference Example 1. Experiment Preparation

1-1. Experimental material

RPMI 1640 medium, fetal bovine serum, penicillin and streptomycin were purchased from Gibco BRL, Grand Island, NY, USA, and MTT ((GrandIsland, NY). 3- (4 , 5-Dimethylthiazol-2-yl) -2,5-diphenyl-tertazoliumbromide) and DMSO (dimethyl sulfoxide) were purchased from Sigma Chemical Co., St. Louis, Mo., USA.

1-2. Preparation of Cancer Cell Lines

The AGS cell line, a human gastric cancer cell used in this experiment, was used by the Korea Biotechnology Research Institute (KCLB, Seoul, Korea). The cell culture was performed in RPMI 1640 or DMEM (Dulbecco's modified Eagle's minimum essential medium). Incubated at 37 ° C. and 5% CO 2 using medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin and streptomycin inactivated by heat treatment.

1-3. Treatment of Black Garlic Concentrate

In order to process the black garlic concentrate of Example 4, AGS cell lines were divided into 6 wells at 1 × 10 ⁵ cells / ml, and stabilized for 24 hours. Microorganisms and impurities were filtered out using a filter unit for syringes with a pore size of 0.22 μm or disposable pump filter units, and then directly treated while grinding the growth medium of the cells.

Reference Example 2. Statistics Processing Method

The measurements were made three times, and the statistical and significance verification of the experimental results was evaluated by Sigma Plot Program 4.0 (SPSS Ins. USA).

Experimental Example 1. Measurement of cell growth inhibition effect of black garlic concentrate

In order to confirm the cytotoxicity of the black garlic concentrate obtained in Example 1, the experiment was performed using the MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay as follows. Carmichael et al., Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemisensitivity testing. Cancer Research, 47 , pp. 944-946, 1987.

The MTT assay is an indirect measure of cell viability. In living cells, water-soluble tetrazolium salt MTT (yellow) is transformed into formazan crystals (purple) by the dehydrogenase action of mitochondria, an intracellular organelle. The basic principle is that the larger the number of living cells, the higher the formazan crystal formation and the higher the absorbance.

The medium treated with black garlic concentrate of Reference Example 1-3 was stabilized for about 24 hours, and then diluted and cultured in a medium. The black garlic concentrate was treated at various concentrations (0, 1, 2, 3 mg / ml) to remove the medium after 48 hours, and 200 μl of MTT solution (5.0 mg / ml) was diluted to 0.5 mg / ml. And incubated for 3 hours. After incubation, remove the MTT reagent and dispense 100 μl of DMSO to dissolve all the generated formazin in the wells, and absorb the absorbance at 540 nm with an ELISA reader (Molecular Devices, Sunnyvale, CA, USA). Measured.

As a result, as shown in FIG. 2, it was found that the AGS cell line inhibited the proliferation of cancer cells depending on the treatment concentration of the black garlic concentrate. In particular, the survival rate of the cells decreased to 43% at the concentration of 3 mg / ml in the black garlic concentrate. By confirming that, as the concentration of the black garlic concentrate increases, various components contained in the black garlic concentrate have a strong anticancer effect as cell viability decreases (see FIG. 2).

[National R & D project supporting this invention]

[Task unique number]

06-RIS-001

[Name of Buddha]

Gyeongnam Provincial Namhae University

[Name of research project]

Core Competency Enhancement Project of Namhae Garlic Industry by Establishing Treasure Island Cluster

[Name of Research Project]

Development of anti-cancer and cancer prevention material from black garlic and development of cancer prevention functional product (Euidae University) / Development of functional product for improving insulin resistance and hyperlipidemia based on Namhae garlic

[Host]

Gyeongnam Provincial Namhae University

[Research period]

April 27, 2009 to June 30, 2009

1 is a view showing a manufacturing process of the black garlic concentrate,

Figure 2 is a diagram showing the cell proliferation inhibitory effect by black garlic concentrate using the MTT assay.

Claims (4)

A first step of fermenting garlic with black garlic; Removing the skin of the black garlic prepared in step 1 and hot water extraction, cold immersion of a solvent selected from 1 to 30 times the volume of the sample weight water, lower alcohol of 1 to 4 carbon atoms or a mixed solvent thereof at room temperature for 1 to 48 hours A second step of extracting by extraction method such as extraction, ultrasonic extraction, reflux cooling extraction, heating extraction, and concentrating under reduced pressure at a temperature of 30 to 100 ° C. to obtain black garlic extract; A third step of mixing plum and sugar at a ratio of 1 to 20: 1 to 20 (w / w / w), fermenting at a temperature of 10 to 100 ° C. for 1 to 6 months, and then filtering to obtain a plum extract; Soybean digested extract was added to the incubator, sterilized, and then inoculated with 50 to 1000 milliliters of mushroom spawn to incubate for 24 to 144 hours to obtain mycelium. Or a fourth step of extracting and filtering the solvent selected from these mixed solvents by extraction methods such as hot water extraction, cold sediment extraction, ultrasonic extraction, reflux cooling extraction, and heat extraction at room temperature for 1 to 48 hours to obtain mushroom mycelium extract; The black garlic extract, plum extract and mushroom mycelium extract prepared above were mixed at a mixing ratio of 50 to 90:10 to 40: 1 to 10 (w / w / w), sterilized, aliquoted and sealed, and then black garlic concentrate. Method for producing a functional black garlic concentrate, characterized in that it comprises a fifth step of obtaining. The method of claim 1, wherein the black garlic is a first step of ripening garlic with hot air for 150 to 500 hours at 10 to 120 ℃; A second step of naturally drying the garlic aged in the step for 10 to 60 hours; The dry garlic further comprises a second aging step of aging for 10 to 100 hours with hot air at a temperature of 10 to 50 ℃ again. The method according to claim 1, wherein the mushroom is mushroom mushroom, matsutake mushroom, shiitake mushroom, agaricus, eggplant mushroom, roe deer mushroom or chaga mushroom. Black garlic concentrate obtained with high nutrition, flavor, palatability and anticancer effect obtained by the method of claim 1.

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