KR20240108834A - Method for cultivation of mung bean leaves with increased isoflavone derivatives and its use - Google Patents

Abstract

The present invention discloses a method of cultivating mung bean leaves with enhanced isoflavone derivatives and their use.
According to the method for cultivating mung bean leaves according to the present invention, mung bean leaves with significantly increased isoflavone derivatives can be cultivated in a short period of time (20 to 40 days). In addition, the cultivation method of the present invention can stably produce mung bean leaves with a high content of isoflavone derivatives throughout the year.
Mung bean leaves, extracts thereof, or fractions thereof enhanced with isoflavone derivatives according to the present invention can be used as an estrogen substitute, and in pharmaceutical compositions, liver preparations, for preventing and improving diseases caused by estrogen deficiency, for antioxidant purposes, and for skin care. It is useful as a material for functional foods and cosmetics.

Description

{Method for cultivation of mung bean leaves with increased isoflavone derivatives and its use}

The present invention relates to a method of cultivating mung bean leaves with enhanced isoflavone derivatives and their use, and more specifically, to a method of cultivating mung bean leaves with enhanced isoflavone derivatives using ethylene treatment under specific conditions, and to a method of cultivating mung bean leaves with enhanced isoflavone derivatives using ethylene treatment under specific conditions. It concerns the use of mung bean leaves.

The female hormone, estrogen, begins to decrease after the age of 40, ultimately causing menopause. When estrogen decreases, aging begins in the bone, heart, skin, and brain areas where this hormone acts. To suppress this aging process, estrogen replacements are urgently needed, and isoflavone derivatives with structural similarity have been reported as effective estrogen replacements.

Isoflavone derivatives are representative flavonoids with excellent pharmacological efficacy, and a lot of capital is being invested worldwide in the development of functional plants containing high levels of these. Isoflavones are non-steroidal estrogens and are called phytoestrogens because they have similar chemical and physiological properties to estrogen. They are substances that have estrogen or anti-estrogen effects by competitively binding to estrogen receptors (Hudson, 2001). They have a similar structure to the female hormone, estrogen, and show useful physiological activities that occur in estrogen. Consumption of isoflavones is known to regulate the endocrine status of women and affect ovarian periodicity (Kenneth, Setchell, & Cassidy, 1999). In particular, it is reported to reduce the risk of osteoporosis, one of the menopausal symptoms that can occur due to a lack of female hormones, and to lower plasma cholesterol levels (Obstetrics & Gynecology 2001. 389). It is also known to have an effect in lowering the risk of coronary heart disease (CAHD) and to have excellent antioxidant activity.

Among isoflavone derivatives, non-glycosides include genistein, daidzein, and glycitein, and glycosides include genistin, daidzin, and glycitin. ), etc. While non-fermented foods derived from soybeans contain many glycosides, fermented foods using soybeans, such as soybean paste, mainly contain non-glycosides as sugars are decomposed from glycosides by bacteria involved in fermentation.

Isoflavone derivatives from legumes (soybeans, mung beans, etc.) have high affinity for the estrogen receptor ER-beta (estrogen receptor beta) and are the most effective estrogen substitutes (Endocrinology 1997, 863).

In this way, mung bean isoflavone derivatives are useful biological materials as estrogen substitutes, but due to the high content of protein, fat, and carbohydrates contained in mung beans, processing processes such as extraction, purification, and commercialization of isoflavone derivatives are difficult. In addition, unlike soybean leaves, mung bean leaves cannot be used as food and are discarded.

Research on crops with a high content of isoflavone derivatives is mainly conducted on soybeans, and a cultivation method technology to produce soybean leaves high in isoflavone derivatives by treating soybean leaves with ethylene or ethephon, an ethylene donor, was recently reported (Korea) Registered Patent No. 10-1451298, Korean Patent No. 10-2282625).

Therefore, there is a need to develop a method of cultivating mung bean leaves with a high content of isoflavone derivatives so that discarded mung bean leaves can be used.

Accordingly, as a result of continuous research to meet the needs of the prior art, the present inventors confirmed that it was possible to cultivate mung bean leaves with significantly increased isoflavone derivatives through ethylene treatment under specific conditions, and completed the present invention.

The purpose of the present invention is to provide a method for cultivating mung bean leaves with enhanced isoflavone derivatives.

Another object of the present invention is to provide mung bean leaves with enhanced isoflavone derivatives grown by the above method.

Another object of the present invention is to provide a health functional food containing mung bean leaves with enhanced isoflavone derivatives.

Another object of the present invention is to provide cosmetics containing mung bean leaves with enhanced isoflavone derivatives.

Another object of the present invention is to provide a pharmaceutical composition comprising mung bean leaves enhanced with the isoflavone derivatives.

According to the object of the present invention, the present invention provides a method for cultivating mung bean leaves enhanced with isoflavone derivatives of the following formula (1):

[Formula 1]

In the above formula, R ₁ is H or OH, and R ₂ is glucose or glucose malonate.

The cultivation method is

i) sowing mung bean seeds or germinated mung beans in a cultivation facility and growing them for 20 to 40 days;

ii) treating the grown mung beans with ethylene at a concentration of 2,500 to 10,000 ppm 2 to 3 times for 8 to 12 hours each; and

iii) harvesting mung bean leaves.

In the present invention, the isoflavone derivative of Formula 1 may be any one or more of the compounds represented by the following [Formula 2] to [Formula 7]:

Step i) Growth

Mung bean seeds or germinated mung beans are sown in a cultivation facility and grown for 20 to 40 days.

In the present invention, 'cultivation facility' is not limited as long as it is a cultivation facility that can artificially control the growth environment, and includes smart farms, plant factories, plant cultivation devices, vertical farms, vertical cultivation devices, container cultivation devices, hydroponic cultivation devices, and smart chambers. , a greenhouse system, a cultivation system, etc., and preferably a vertical plant factory or a smart chamber.

For example, a vertical plant factory includes a plant factory section including an internal space; A door part installed in the center of the plant factory part; LED lighting facilities, cooling and heating facilities for temperature control, ventilation facilities for plant and factory ventilation; irrigation facilities; ICT facilities for remotely automated systems; and an operating application (FIGS. 1A and 1B).

In another example, a smart chamber includes a chamber part including an internal space; A door part that opens and closes the chamber part; an observation window installed in the center of the door; An LED light source installed on the ceiling inside the chamber; Temperature, humidity and carbon dioxide meters; Ventilation fan for chamber ventilation; Temperature control fan installed on the back inside the chamber unit; ethylene gas injector; ethylene gas meter; A light source control device that controls the LED light source; Automatic temperature and humidity control and data collection device; water circulator for temperature control; ethylene gas cylinder; and a data storage device (FIGS. 2A and 2B).

In step i), mung bean seeds or germinated mung beans can be used for sowing, and preferably sprouted mung beans are sowed.

The mung beans germinated in the present invention are made by adding 1 to 3 times the weight of the mung bean seed in water, soaking the mung beans at 15 to 25 ℃, placing them in a germination machine or bean sprout maker at 20 to 30 ℃, and germinating them in a dark room for 16 to 24 hours. It can be manufactured as

In step i), the growth of mung beans is carried out 20 to 40 days after sowing, preferably 30 to 35 days. If it is less than 20 days, the biomass content of mung beans is low, and if it is more than 40 days, problems occur in growth in the cultivation facility.

For mung bean growth, the LED light source installed in the cultivation facility uses a combination of blue and red dual mixed light sources and white plant growth LEDs.

The amount of light (Lux) in the cultivation facility for growing mung beans is about 10,000 to 15,000. If the temperature is less than 10,000 Lux, photosynthesis is not smooth, so the mung bean biomass (leaf) content is low and the production of isoflavone derivatives is not smooth. If it is more than 15,000 Lux, growth is not smooth and leaves die due to the high amount of light. .

The temperature in the cultivation facility for growing mung beans is preferably 20 to 35°C, and more preferably 25 to 30°C. If the temperature is below 20℃, mung beans cannot grow due to cold damage, and if it is above 35℃, high temperature failure may occur.

The humidity in the cultivation facility for growing mung beans is preferably 40 to 80%, and more preferably 50 to 60%. If the moisture content is less than 40%, the photosynthetic metabolism of the mung bean may not be smooth due to lack of moisture, and the leaves may die, and if it exceeds 80%, the roots may rot due to excessive moisture.

Step ii) Ethylene treatment

The grown mung beans are treated with ethylene at a concentration of 2,500 to 10,000 ppm 2 to 3 times for 8 to 12 hours each.

The ethylene treatment concentration is a high concentration of 2,500 to 10,000 ppm, preferably 5,000 to 1,0000 ppm. When the ethylene treatment concentration is less than 2,500 ppm, there is almost no accumulation of isoflavone derivatives in mung bean leaves, and even if the ethylene treatment concentration exceeds 10,000 ppm, there is no significant difference in the accumulation of isoflavone derivatives.

The preferred ethylene treatment time is 8 to 12 hours. If the ethylene treatment time is less than 8 hours, isoflavone derivative accumulation is not sufficient, and if the ethylene treatment time is more than 12 hours, there is no significant difference in isoflavone derivative accumulation.

Ethylene treatment is preferably performed 2 to 3 times. If ethylene treatment is less than two times, isoflavone derivative accumulation is not sufficient, and if ethylene treatment is more than three times, there is no significant difference in isoflavone derivative accumulation.

In step ii), the ethylene treatment takes place in a closed space with the cultivation facility completely closed. In other words, the door to the cultivation facility is closed, the ventilation fan is blocked, and the ethylene is treated.

The time between ethylene treatment and the next ethylene treatment is about 1 to 2 hours, and the cultivation facility is opened and ventilated to remove ethylene and stabilize the mung beans.

Before ethylene treatment, the mung beans are kept in the closed space as above for 1 to 2 hours to stabilize. It is stabilized to control humidity due to the moisture generated by the respiration of mung beans.

During ethylene treatment, the cultivation facility uses a single light source of white, blue, and red; Dual mixed light sources of white and blue, white and red, and blue and red; Alternatively, a triple mixed light source of white, blue, and red can be used, and a triple mixed light source of white, blue, and red is preferred, and the light quantity is 100 ㎛ol/m ² /s or more, preferably 200 ㎛ol/s. Use m ² /s or more.

During ethylene treatment, the temperature of the cultivation facility is preferably 20 to 40°C, and more preferably 30 to 35°C. The humidity is preferably 60 to 90%, and more preferably 70 to 80%. If the ethylene treatment temperature is less than 20℃ and humidity less than 60%, the metabolism of mung beans is not smooth, so isoflavone derivatives are hardly accumulated, and if the temperature is over 40℃ and humidity exceeds 90%, metabolic processes such as respiration are also impaired due to the high temperature and excessive moisture content. This is not smooth and isoflavone derivatives are not made.

iii) Harvest

Harvest the mung bean leaves on which ethylene treatment has been completed in step ii).

As described above, the cultivation method according to the present invention can significantly increase the content of isoflavone derivatives in mung bean leaves by treating mung beans with ethylene under specific conditions such as step ii) before harvesting.

In the case of mung bean leaves grown according to the present invention (about 14,333 μg/g; Example 1), mung bean leaves grown in a vertical plant factory and not treated with ethylene (about 1,016 μg/g; Comparative Example 3), germinated mung beans Compared to (about 48 μg/g; Comparative Example 2) and mung bean seeds (about 1.8 μg/g; Comparative Example 1), the content of isoflavone derivatives was significantly increased by about 14.1 times, about 298.63 times, and about 7,962 times, respectively ( Figure 10).

According to another object of the present invention, there is provided mung bean leaves enhanced with isoflavone derivatives grown by the above method.

Mung bean leaves enhanced with isoflavone derivatives according to the present invention contain more than 14,333 μg/g of isoflavone derivatives, more than 13,347 μg/g of beta-glycoside isoflavone derivatives, and more than 13,347 μg/g of maloyl-beta-glycoside isoflavone derivatives. It contains more than 324 μg/g of flavone derivatives and more than 761 μg/g of aglycone isoflavone derivatives (Table 7, Figure 10).

Mung bean leaves enriched with isoflavone derivatives according to the present invention can be used as a material for foods, cosmetics, and pharmaceutical compositions by themselves or by extracting them with a solvent.

In the present invention, water, alcohol, or a mixture thereof can be used as the extraction solvent. It is preferable to use C1 to C2 lower alcohol as the alcohol, and it is preferable to use ethanol or methanol as the lower alcohol.

The extraction method is preferably shaken extraction, Soxhlet extraction, or reflux extraction, but is not limited thereto. Specifically, it is preferable to extract by adding 1 to 10 times (v/w) of the extraction solvent to dried mung bean leaves, the extraction temperature is preferably 20 to 100°C, and the extraction time is preferably 10 to 48 hours. However, it is not limited to this.

The extract can be concentrated under reduced pressure and then dried. It is preferable to use a vacuum concentrator or a vacuum rotary evaporator for reduced pressure concentration, but is not limited thereto. In addition, drying is preferably performed by reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

The extract can be further fractionated with an organic solvent, and the extract can be suspended in water and then fractionated using n-hexane, chloroform, ethyl acetate, or butanol as an organic solvent, with chloroform being preferred. Fractions can be obtained from the extract by repeating the fractionation process 1 to 5 times, preferably 3 times, and it is preferable to concentrate under reduced pressure after fractionation, but is not limited to this.

According to another object of the present invention, the present invention provides a health functional food containing mung bean leaves with enhanced isoflavone derivatives.

The health functional food according to the present invention is useful for preventing and improving diseases caused by estrogen deficiency, as an antioxidant, and for skin care.

In the present invention, the disease caused by estrogen deficiency is any one selected from the group consisting of osteoporosis, heart disease, breast cancer, vulvovaginal disease, hyperlipidemia, skin aging, and facial flushing.

Since isoflavone derivatives have established physiological activity for preventing and improving diseases caused by estrogen deficiency as described above, for antioxidant purposes, and for skin care, the mung bean leaves according to the present invention containing a high content of isoflavone derivatives are It can be usefully used as a health functional food as described above.

The health functional food according to the present invention may contain mung bean leaves with enhanced isoflavone derivatives as is, or may be included together with other foods or food ingredients in the form of extracts or fractions as described above.

In the present invention, 'health functional food' refers to food manufactured using nutrients that are easily deficient in daily diet or raw materials or ingredients with useful functions for the human body, and helps maintain the health of the human body.

In the present invention, the form and type of health functional food are not particularly limited. Specifically, it may be in the form of tablets, capsules, powders, granules, liquids, and pills. Nutraceutical foods may contain various flavors, sweeteners or natural carbohydrates as additional ingredients. The sweetener may be a natural or synthetic sweetener, and examples of natural sweeteners include thaumatin and stevia extract. Meanwhile, examples of synthetic sweeteners include saccharin and aspartame. Additionally, natural carbohydrates may include monosaccharides, disaccharides, polysaccharides, oligosaccharides, and sugar alcohols.

In the present invention, in addition to the above additional ingredients, the health functional food includes nutrients, vitamins, electrolytes, flavors, colorants, pexane and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, and glycerin. , may further contain alcohol, etc. These ingredients can be used independently or in combination.

In the present invention, the type of food is not particularly limited and may include pills, tablets, capsules, tea, snacks, flour foods, soups, dairy products, sauces, meat additives, beverages (pouches, drinks, etc.), etc., but is limited thereto. It doesn't work.

Preferably, the food according to the present invention may be tea. The tea according to the present invention may be a mixed tea mixed with mung bean leaves enhanced with the isoflavone derivative according to the present invention and herbal plants with flavor. The herbal plant is preferably at least one selected from the group consisting of chamomile, lemongrass, rosehip, lavender, peppermint, fennel, rosemary, jasmine, hibiscus, roseflower, apple fruit, strawberry fruit, lemon fruit and orange flower. The tea or mixed tea of the present invention may be processed into a form selected from the group consisting of flavored tea, tea bags, instant tea, and cans.

According to another object of the present invention, the present invention provides a cosmetic product comprising mung bean leaves enhanced with the above isoflavone derivatives.

The cosmetic according to the present invention may contain mung bean leaves with enhanced isoflavone derivatives as is, or may be included together with other cosmetic ingredients in the form of extracts or fractions as described above.

In the present invention, cosmetics may be in the form of lotions, ointments, gels, creams, patches, mask packs or sprays, but are not limited thereto, and may further include fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, etc. Antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, It may contain any additives commonly used in cosmetics, such as pigments, hydrophilic or lipophilic active agents, and lipid vesicles.

According to another object of the present invention, the present invention provides a pharmaceutical composition for the prevention and treatment of diseases caused by estrogen deficiency, comprising mung bean leaves with enhanced isoflavone derivatives.

In the present invention, the disease caused by estrogen deficiency is any one selected from the group consisting of osteoporosis, heart disease, breast cancer, vulvovaginal disease, hyperlipidemia, skin aging, and facial flushing.

Since isoflavone derivatives are established as estrogen substitutes for the prevention and improvement of diseases caused by estrogen deficiency as described above, the mung bean leaves enhanced with isoflavone derivatives according to the present invention containing a high content of isoflavone derivatives are as described above. It is useful as a pharmaceutical composition for the same purpose.

The pharmaceutical composition according to the present invention can be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. You can. Carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, and methyl cellulose. , microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations are mixed herbal medicines with at least one excipient, such as starch, calcium carbonate, and sucrose. Alternatively, it is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

The pharmaceutical composition according to the present invention can be administered orally or parenterally. When administered parenterally, it can be administered externally under the skin or by intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. You can.

The preferred dosage of the pharmaceutical composition according to the present invention varies depending on the patient's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. For a desirable effect, the composition is preferably administered at 0.0001 to 1 g/kg per day, preferably at 0.001 to 200 mg/kg. The above administration may be administered once a day, or may be administered several times.

According to the method for cultivating mung bean leaves according to the present invention, mung bean leaves with significantly increased isoflavone derivatives can be cultivated in a short period of time (20 to 40 days). In addition, the cultivation method of the present invention can stably produce mung bean leaves with a high content of isoflavone derivatives throughout the year.

Mung bean leaves, extracts thereof, or fractions thereof enhanced with isoflavone derivatives according to the present invention can be used as an estrogen substitute.

Mung bean leaves with enhanced isoflavone derivatives, extracts thereof, or fractions thereof according to the present invention are used as ingredients for pharmaceutical compositions, liver functional foods, and cosmetics for preventing and improving diseases caused by estrogen deficiency, for antioxidant purposes, and for skin care. useful.

Figure 1a shows an example of a configuration diagram of a vertical plant factory, a type of cultivation facility according to the present invention. Figure 1b shows an example of the internal facilities and automatic control system of a vertical plant factory. Figure 1c shows mung beans grown 15 and 35 days after sowing in a vertical plant factory.
Figure 2a shows an example of the configuration of a smart metabolite chamber, a type of cultivation facility according to the present invention. Figure 2b is a photograph showing the actual manufactured product of the smart metabolite chamber. Figure 2c shows mung beans before and after ethylene treatment.
Figure 3a is an HPLC chromatogram analyzing isoflavone derivatives of mung bean seeds (Comparative Example 1), Figure 3b is an HPLC chromatogram analyzing isoflavone derivatives of germinated mung beans (Comparative Example 2), and Figure 3c is a vertical plant. This is an HPLC chromatogram analyzing the isoflavone derivatives of mung bean leaves grown 35 days after sowing in a factory (Comparative Example 3), and Figure 3d shows a sample grown in a vertical plant factory for 35 days after sowing and treated with ethylene in a smart metabolite chamber. This is an HPLC chromatogram analyzing isoflavone derivatives of mung bean leaves (Example 1).
a is daidzein, b is genistin, c is malonyl daidzine, d is malonyl genistin, e is daidzein, f is genistein.
Figure 4 shows the BPI spectrum of LC-ESI-TOF/MS of peak a in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, daidzine.
Figure 5 shows the BPI spectrum of LC-ESI-TOF/MS of peak b in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, genistin.
Figure 6 shows the LC-ESI-TOF/MS BPI spectrum of peak c in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, malonyl daidzine.
Figure 7 shows the LC-ESI-TOF/MS BPI spectrum of peak d in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, malonyl genistin.
Figure 8 shows the BPI spectrum of LC-ESI-TOF/MS of peak e in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, daidzein.
Figure 9 shows the BPI spectrum of LC-ESI-TOF/MS of peak f in the HPLC chromatogram of Figure 3d and the chemical formula of the corresponding compound, genistein.
Figure 10 is a graph comparing the isoflavone derivative contents of mung bean leaves of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Example 1.
Figure 11 is a graph comparing the beta-glycoside isoflavone content of mung bean leaves and ethylene-treated soybean leaves according to the present invention.

Hereinafter, the present invention will be described in detail with reference to drawings, examples and manufacturing examples. However, the drawings, examples, and manufacturing examples only illustrate the present invention, and the content of the present invention is not limited by the drawings, examples, and manufacturing examples.

<Vertical plant factory>

The vertical plant factory used for growing mung beans in the present invention will be described in detail with reference to the drawings.

Figure 1a shows an example of a configuration diagram of a vertical plant factory, a type of cultivation facility according to the present invention.

Referring to Figure 1a, the external width × length × height of the vertical plant factory according to the present invention may be 3000 mm × 8200 mm × 3000 mm, and the internal width × length × height may be 2800 mm × 7700 mm × 2720 mm. is desirable.

A plant factory consists of a plant factory section including an internal space, a door installed in the center of the plant factory section, an LED lighting facility, an air-conditioning/heating facility for temperature control, a ventilation facility for ventilation of the plant factory section, an irrigation facility, and a remotely automated system. Includes ICT facilities and operating apps for

The configuration of the vertical plant factory may include LED lighting facilities, cooling/heating facilities, ventilation facilities, irrigation facilities, ICT facilities, and operating apps. The LED lighting facility acts like sunlight, and unlike general LED lighting, it is an LED for plant growth and can be adjusted to suit the cultivation environment.

The cooling and heating facilities control temperature and humidity and can control temperature and humidity by using data programs accumulated according to the external temperature and humidity.

The ventilation facility is a facility that allows plants to breathe, and through ventilation, oxygen moves away and carbon dioxide necessary for photosynthesis enters the stomata.

The irrigation facility is a facility for rain and watering, and can be used easily and conveniently by automatically watering the desired amount at the desired time.

The above ICT equipment is a facility that automatically controls temperature, humidity, and ventilation, etc. remotely, and plays an important role in increasing leisure time and improving quality of life by being free from the temporal/spatial constraints of agricultural work.

The operation application is a remotely controllable application that controls the growth environment remotely via mobile instead of directly in the field and analyzes crop growth information using big data.

<Smart-metabolite chamber>

The smart metabolite chamber used in the ethylene treatment step in the present invention will be described in detail with reference to the drawings.

Figure 2a shows an example of the configuration of a smart metabolite chamber, a type of cultivation facility according to the present invention.

Referring to Figure 2a, the smart-metabolite chamber according to the present invention includes a chamber part including an internal space, an opening and closing door part that opens and closes the chamber part, an observation window 13 installed in the center of the door part, and a ceiling inside the chamber part. Installed LED light source (3), temperature, humidity and carbon dioxide meter (5); Ventilation fan (6) for ventilation of the chamber, temperature control fan (7) installed on the back of the chamber, ethylene gas injection device (10), ethylene gas meter (11), light source control device (1, 2), it may include an automatic temperature and humidity control and data collection device (4), a water circulation device for temperature control (8), an ethylene gas cylinder (9), and a data storage device (12).

In the smart-metabolite chamber according to the present invention, the chamber portion may have a size of 500 to 1200 mm x 400 to 1000 mm x 400 to 8000 mm in width x length x height, and the size of 700 mm x 500 mm x 500 mm is preferred. The chamber part is preferably made of aluminum, stainless steel, wood, or polyacrylic plate, but is more preferably made of aluminum or stainless steel.

The door part allows the chamber part to be opened and closed, and an observation window 13 is installed in the center so that the growth of the plant can be observed (FIG. 2c). The observation window 13 may be made of a material such as transparent acrylic plate or glass.

The LED light source unit 3 can be installed on the ceiling inside the chamber and can irradiate plants with one or more types of light selected from the group consisting of red, blue, white, UV-A, and UV-B. The amount of light can be adjusted from 1% to 100% for each LED light source under dark room conditions.

The light source control device (1, 2) that controls the LED light source can be installed on any part of the outer surface of the chamber, and is a three-channel light source control device (1) of red (660 nm), blue (450 nm), and white. It can be divided into UV-A (375 nm) and UV-B (315 nm) light source control device (2). The light source control device can control the light amount of each LED light source from 1% to 100%.

The temperature, humidity and carbon dioxide meter 5 can be installed on one side of the chamber and is connected to the temperature and humidity automatic control and data collection device 4 installed outside the chamber. Temperature can be measured from -10℃ to 60℃, and humidity can be measured from 0% to 100%. Carbon dioxide can be measured from 0 to 2,000 ppm.

One or more ventilation fans 6 may be installed to ventilate the chamber portion.

The temperature control fan (7) is installed at the rear of the chamber, functions to control the temperature of the internal space of the chamber, and is connected to a water circulation device (8) for temperature control installed outside. The temperature within the smart-metabolite chamber can be maintained constant from 15℃ to 45℃, and the actual humidity within the smart-metabolite chamber can be maintained constant from 20% to 100%.

The ethylene gas injection device 10 is installed on one side of the chamber to spray ethylene on plants, and is connected to an ethylene gas cylinder 9 installed outside the chamber. The ethylene gas concentration inside the chamber can be sprayed at a concentration of up to 10,000 ppm, and the ethylene gas is evenly sprayed into the smart-metabolite chamber through the spray device 10 that temporarily passes through water. Ethylene gas injection is controlled by installing a gas switch.

The ethylene gas meter 11 is installed below the temperature control fan 7 to measure the ethylene gas concentration inside the chamber. The ethylene gas meter is installed externally and measures the amount detected by the meter.

The data storage device 12 is installed outside the chamber and is connected to the automatic temperature and humidity control and data collection device 4. Data collection device 4 may be a computer.

The smart metabolite chamber includes a power supply (DC24VDC), a voltage drop module (VDIV10.1), a modem (Ethernet modem) for remote access to the data logger, and a wired/wireless router for wireless Wi-Fi modem communication. Electrical controllers for operating the in-router and controlled fan/ventilation fan may be additionally installed.

Figure 2b is a photograph showing the actual manufactured product of the smart metabolite chamber according to the present invention.

Reference Example 1: Preparation of extract

The extract for measuring isoflavones was dried and powdered, and 10 g of each powdered harvest was added to 400 ml of 50% methanol and extracted at 25°C for 5 days. The extracted solvent was concentrated under reduced pressure to obtain an extract with a yield of 16-20%.

Reference Example 2: Isoflavone derivative analysis

High performance liquid chromatography (HPLC) analysis was performed to confirm the degree of accumulation of isoflavone derivatives in the extract.

The extract prepared as in Reference Example 1 was centrifuged at 3500 rpm for 10 minutes using a centrifuge, and the supernatant was filtered using a 0.45 ㎛ syringe filter. The instrument used for isoflavone analysis was HPLC using the Agilent 1260 system LC from Agilent Technologies Inc. (Waldbronn, Germany), and the column was Lichrophore 100 RP C18 column (4.6×250 mm, 5μm). , Merck, Germany) and DAD (diode array detector) was used. As for the analysis conditions, 20 ㎕ of sample extract was injected, the column temperature was 30°C, the solvent was flowed at a flow rate of 1 ml/min, and the analysis solvent was 0.2% acetic acid aqueous solution (solvent A) with 0.2% acetic acid added. Isoflavone derivatives were separated by applying gradient elution as shown in Table 1 using a mixed solution of acetonitrile (solvent B) and confirmed at a detection wavelength of 254 nm. The standard solution used for quantitative analysis was prepared by dissolving 98% purity daidzin, genistin, daidzein, and genistein purchased from Sigma (USA) in methanol in the range of 1 to 100 ㎍/ml. HPLC analysis was performed with the prepared standard solution, and a calibration curve was created from the peak area.

time (minutes) Solvent A (%) Solvent B (%) 0 100 0 15 90 10 25 80 20 35 75 25 45 65 35 50 65 35

Test Example 1: Analysis of optimal conditions for ethylene treatment of mung bean leaves

<Ethylene treatment concentration>

Add about twice the weight of the mung bean seed with water, hydrate it at 20℃ for 15 hours, put it in a germination device in a dark room, and germinate it at 20℃ for 24 hours. The germinated mung bean (germinated mung bean) is placed in a plant pot for general horticulture. After filling about 80 g of topsoil, 2 to 3 seeds were sown. The pot was then grown in a vertical plant factory for 35 days. At this time, in the plant factory environment, the amount of light was set to about 13000 LUX and about 1200 FC, the temperature was set to 25 ± 0.5°C, and the humidity was set to 65 ± 5%. The grown mung beans were divided into 4 groups, transferred to a smart metabolite chamber, and ethylene treatment was performed in a completely closed, sealed space. After completely blocking the entrance and ventilation fan of the smart metabolite chamber, create a closed space. The light source control device (1) sets the white, blue, and red light ratio to 1:1:1 and the light amount is 400 ㎛ol/m. ² /s and stabilized for 2 hours, grown at a temperature of 35 ± 1°C and humidity of 80 ± 5%, then ethylene gas was injected at 0, 2,500, 5,000 and 10,000 for each group through the ethylene gas injection device (9). Harvested after treatment for 24 hours at a concentration of ppm. Extracts were prepared from the harvested mung bean leaves of each group as in Reference Example 1, and the isoflavone derivative content was analyzed in the same manner as in Reference Example 2. The results are shown in Table 2.

content
(㎍/g dw) Treatment by ethylene concentration (ppm) 0 2500 5000 10000 β-Glycosides Daidzin (a) 1402.71 2861.59 3461.06 3973.82 Genistin (b) 290.51 782.72 953.20 953.88 Sum 1,693.22 3,644.31 4,414.26 4,927.70 Malonly-β-Glycosides Malonlydaidzin (c) 0 0 0 0 Malonlygenistin (d) 128.21 91.37 118.15 83.68 Sum 128.21 91.37 118.15 83.68 Aglycones Daidzein (e) 170.88 240.01 304.56 327.28 Genistein (f) 22.84 55.80 67.18 65.35 Sum 193.72 295.81 371.74 392.63 Note 1) All experiments were performed 5 times and expressed as the average value.
Note 2) Ethylene treatment was performed once for 24 hours at 35 degrees at each concentration.

As shown in Table 2, the isoflavone derivative content of mung bean leaves treated with different ethylene concentrations (0, 2,500, 5,000, and 10,000 ppm) increased as the ethylene concentration increased. When comparing beta-glycoside content by concentration, compared to 0 ppm ethylene treatment, it was found that treatment with 2,500 ppm increased by 2.2 times, 2.6 times with 5,000 ppm treatment, and 2.9 times with 10,000 ppm treatment.

According to the above results, the appropriate concentration of ethylene treatment for mung bean leaves is 2,500 to 10,000 ppm, preferably 5,000 to 10,000 ppm.

<Ethylene treatment time>

Mung beans were grown as described above, divided into 4 groups, and harvested after treating each group with ethylene at a concentration of 10,000 ppm for 0, 12, 24, and 36 hours in a smart metabolite chamber in the same environment as above. After preparing the extract from mung bean leaves as in Reference Example 1, the isoflavone derivative content was analyzed in the same way as in Reference Example 2, and the results are shown in Table 3.

content
(㎍/g dw) Ethylene processing time (hours) 0 12 24 36 β-Glycosides Daidzin (a) 552.81 887.11 3393.15 3772.85 Genistin (b) 130.47 198.56 696.59 1186.42 Sum 683.28 1,085.67 4,089.74 4,959.27 Malonly-β-Glycosides Malonlydaidzin (c) 0 0 0 0 Malonlygenistin (d) 0 106.67 69.44 96.03 Sum 0 106.67 69.44 96.03 Aglycones Daidzein (e) 34.81 80.13 351.18 300.76 Genistein (f) 0 11.04 59.95 77.01 Sum 34.81 91.17 411.13 377.77 Note 1) All experiments were performed 5 times and expressed as the average value.
Note 2) Ethylene treatment was performed once at a concentration of 10,000 ppm at 35 degrees for 12, 24, and 36 hours.

As shown in Table 3, as ethylene treatment time increased, the isoflavone derivative content also increased. The total beta-glycoside content was found to be 683.28 ㎍/g at 0 hours. In comparison, the content increased to 1,085.67, 4,089.74, and 4,959.27 ㎍/g at 12, 24, and 36 hours of treatment, respectively, especially at 12 hours. The content rapidly increased by about 3.9 times in 24 hours.

According to the above results, the appropriate ethylene treatment time for mung bean leaves is 24 to 36 hours, most preferably 24 hours.

<Number of ethylene treatments>

Mung beans were grown as above, divided into 4 groups, and treated with ethylene at a concentration of 10,000 ppm in a smart metabolite chamber in the same environment as above. The total time for each group was 0 times, 1 time (24 hours), and 2 times based on 24 hours. They were harvested after treatment once (12 hours each) and three times (8 hours each), and extracts were prepared from the harvested mung bean leaves of each group as in Reference Example 1, and then the isoflavone derivative content was tested in the same manner as in Reference Example 2. was analyzed and the results are shown in Table 4.

content
(㎍/g dw) Number of ethylene treatments (times) 0 One 2 3 β-Glycosides Daidzin (a) 875.69 4,292.08 10633.91 11337.36 Genistin (b) 192.56 1063.76 2268.24 2244.99 Sum 1,068.25 5,355.84 12,902.15 13,582.35 Malonly-β-Glycosides Malonlydaidzin (c) 0 0 189.57 122.60 Malonlygenistin (d) 0 124.09 262.07 237.88 Sum 0 124.09 451.64 360.48 Aglycones Daidzein (e) 65.12 426.58 831.27 678.62 Genistein (f) 8.49 68.15 121.97 90.59 Sum 73.61 494.73 953.24 769.21 Note 1) All experiments were performed 5 times and expressed as the average value.
Note 2) Ethylene treatment was performed once for 24 hours, twice for 12 hours, and three times for 8 hours at 35 degrees at a concentration of 10,000 ppm.

As shown in Table 4, as the number of ethylene treatments increased, the isoflavone derivative content also increased. When compared based on the total beta-glycoside content, it was found to be 1,068.25, 5,355.84, 12,902.15, and 13,582.35 ㎍/g for 0, 1, 2, and 3 treatments, respectively, especially for 2 and 3 treatments, and 1 treatment. It was found that the content of isoflavone derivatives increased significantly compared to the time.

According to the above results, it is desirable to divide the ethylene treatment of mung bean leaves into 2 to 3 treatments.

Manufacturing Example 1: Cultivation of green beans in a vertical plant factory

Mung bean seeds (Comparative Example 1) Approximately twice the weight of water was added, hydrated at 20°C for 15 hours, placed in a germinator in a dark room, and germinated mung beans (germinated mung beans) were prepared by germination at 20°C for 24 hours (Comparison) Example 2).

A plant pot was filled with about 80 g of general horticultural soil, and then 2 to 3 sprouted mung beans were sown. Thereafter, the pots of mung bean seeds sown above were grown in a vertical plant factory for 35 days (Comparative Example 3). At this time, in the plant factory environment, the amount of light was set to about 13000 LUX and about 1200 FC, the temperature was set to 25 ± 0.5 ℃, and the humidity was set to 65 ± 5% (Figures 1a to 1c).

The mung bean plants grown above were transferred to a smart metabolite chamber and ethylene treatment was performed in a completely closed and closed space. After completely blocking the entrance and ventilation fan of the smart metabolite chamber, create a closed space. The light source control device (1) sets the white, blue, and red light ratio to 1:1:1 and the light amount is 400 ㎛ol/m. It was set to ² /s and stabilized for 2 hours at a temperature of 35 ± 1°C and humidity of 80 ± 5%, and then treated with ethylene gas at a concentration of 10,000 ppm a total of 3 times for 8 hours through the ethylene gas injection device (9). Then, the mung beans were harvested (Example 1; FIGS. 2A to 2C). The time between ethylene treatment and the next ethylene treatment is about 2 hours, and the cultivation facility is opened and ventilated to remove ethylene and stabilize the mung beans.

The mung bean seeds of Comparative Example 1, the germinated mung beans of Comparative Example 2, the mung bean leaves of Comparative Example 3, and the mung bean leaves of Example 1 were each washed three times in running water, dried at 50 ± 2°C for 24 to 48 hours, and then powdered. .

Test Example 2: Isoflavone HPLC chromatogram analysis

The powders of Comparative Example 1, Comparative Example 2, Comparative Example 3, and Example 1 prepared in Preparation Example 1 were prepared as extracts as in Reference Example 1, and then HPLC chromatograms of each isoflavone derivative as in Reference Example 2. The analysis results are shown in Figures 3A to 3D.

As shown in FIGS. 3A to 3D, in Comparative Examples 1 and 2 (mung bean seeds and germinated mung beans), the isoflavone derivative peak was almost low or not detected (FIGS. 3A and 3B). In the case of ethylene-untreated mung bean leaves in Comparative Example 3 (mung bean leaves grown in a plant factory), peaks of six isoflavone derivatives (daedzin, genistin, malonyl daidzin, malonyl genistin, daidzein, and genistein) (Figure 3c), and in Example 1 (ethylene-treated mung bean leaves according to the present invention), it can be confirmed that the peaks of six types of isoflavone derivatives are significantly increased (Figure 3d). In particular, the mung bean leaves of Example 1 were found to have significantly increased peak areas of daidzin (peak a) and genistin (peak b).

Test Example 3: Separation, purification and structural analysis of isoflavone derivatives

UPLC-ESI-TOF/MS (Acquity UPLC system coupled with SYNAPT G2-Si HDMS, Waters) analysis was performed on the isoflavone derivatives corresponding to each peak of the mung bean leaves of Example 1 (FIG. 3d).

Specifically, for the extract prepared as in Reference Example 1, a UPLC BEH C18 column (2.1 mm (Solvent B) Isoflavone derivatives were separated by applying gradient elution as shown in Table 5 with the mixed solution. At this time, the amount of extraction solution injected was 1 ㎕, and UPLC-ESI-TOF mass spectrometry was performed under the conditions shown in Table 6, and the results are shown in Figures 4 to 9.

time (minutes) Solvent A (%) Solvent B (%) 0 7 3 5 85 15 10 75 25 11 70 30 12 55 45

Detail condition Ionization ESI-negative mode Capillary voltage 3 kV Sample cone voltage 40V Desolvation gas flow 800 L/h Cone gas flow 30 L/h Desolvation temperature 400℃ Ion source temperature 100℃ Data collection m/z 50-1500 rage with a scan time of 0.2 s Lock mass Leucine-enkephalin (556.2771 Da)

As shown in Figure 4, in the mass spectrometry spectrum of peak a in Figure 3d, [M+H] ^- was found to be 415.0 (theoretical molecular weight: 416.3). From this, it was confirmed that the elemental composition was C ₂₁ H ₂₁ O ₉ , which matched the elemental composition of Daidjin. Peak a matches daidzine shown in the reference (J. Agric. Food Chem. 2016, 64, 7315-7324) and was identified as daidzine of Formula 2.

As shown in Figure 5, in the mass spectrometry spectrum of peak b in Figure 3d, [M+H] ^- was found to be 431.1 (theoretical molecular weight: 432.3). From this, it was confirmed that the elemental composition was C ₂₁ H ₂₁ O ₁₀ , which was consistent with the elemental composition of genistin. Peak b is consistent with genistin shown in the reference (J. Agric. Food Chem. 2016, 64, 7315-7324) and was identified as genistin of formula 3.

As shown in Figure 6, in the mass spectrometry spectrum of peak c in Figure 3D, [M+H] ^- was found to be 501.4 (theoretical molecular weight: 502.4). From this, it was confirmed that the elemental composition was C ₂₄ H ₂₃ O ₁₂ , which was consistent with the elemental composition of malonyl daidzine. Peak c corresponds to malonyl daidzine shown in the reference (J. Agric. Food Chem. 2016, 64, 7315-7324) and was identified as malonyl daidzine of Formula 4.

As shown in FIG. 7, in the mass spectrometry spectrum of peak d in FIG. 3D, [M+H] ^- was found to be 517.1 (theoretical molecular weight: 518.4). From this, it was confirmed that the elemental composition was C ₂₄ H ₂₃ O ₁₃ , which was consistent with the elemental composition of malonyl genistin. Peak d corresponds to malonyl genistin shown in the reference (J. Agric. Food Chem. 2016, 64, 7315-7324) and was identified as malonyl genistin of formula 5.

As shown in Figure 8, in the mass spectrometry spectrum of peak e in Figure 3D, [M+H] ^- was found to be 253.1 (theoretical molecular weight: 254.2). From this, it was confirmed that the elemental composition was C ₁₅ H ₁₀ O ₄ , which was consistent with the elemental composition of malonyl daidzine. Peak e was consistent with malonyl daidzine shown in the reference (Food Chem. 2020, 305, 125462) and was identified as malonyl daidzine of Formula 6.

As shown in Figure 9, in the mass spectrometry spectrum of peak f in Figure 3d, [M+H] ^- was found to be 269.0 (theoretical molecular weight: 270.4). From this, it was confirmed that the elemental composition was C ₁₅ H ₁₀ O ₅ , which was consistent with the elemental composition of malonyl genistin. Peak f was consistent with malonyl genistin shown in the reference (Food Chem. 2020, 305, 125462) and was identified as malonyl genistin of formula 7.

Test Example 4: Content analysis and comparison of isoflavone derivatives

Each extract of Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3 prepared in the same manner as in Reference Example 1 was filtered using a 0.45㎛ syringe filter and then filtered with isoflavones in the same manner as in Reference Example 2. The content was analyzed and the total amount was quantified and compared, and the results are shown in Table 7 and Figure 10.

content
(㎍/g dw) Comparative Example 1 Comparative Example 2 Comparative Example 3 Example 1 β-Glycosides Daidzin (a) 0 43.90 810.69 11,302.36 Genistin (b) 0 0 141.68 1944.66 Sum 0 43.90 952.37 13,247.02 Malonly-β-Glycosides Malonlydaidzin (c) 0 0 0 110.86 Malonlygenistin (d) 0 0 0 214.12 Sum 0 0 0 324.98 Aglycones Daidzein (e) 1.78 4.19 56.32 668.99 Genistein (f) 0 0 7.46 92.24 Sum 1.78 4.19 63.78 761.23 Note 1) All experiments were performed 5 times and expressed as the average value.
Note 2) Ethylene treatment was performed three times for 8 hours at 35 degrees at a concentration of 10,000 ppm.

As shown in Table 7 and Figure 10, most isoflavone derivatives were not detected in the mung bean seeds and germinated mung beans of Comparative Examples 1 and 2, and in the case of the ethylene-untreated mung bean leaves of Comparative Example 3, isoflavone derivatives were not detected. was not detected or was detected in small amounts. In comparison, the mung bean leaves of Example 1 contained 14,333.23 ㎍/g of total isoflavone derivatives, 13,347.02 ㎍/g of beta-glycoside isoflavone derivatives, and 324.98 ㎍/g of maloyl-beta-glycoside isoflavone derivatives. It was confirmed to contain ㎍/g and 761.23 ㎍/g of aglycone isoflavone derivative.

The isoflavone derivative content of Example 1 was significantly increased by about 14.1 times, about 298.63 times, and about 7,962 times compared to Comparative Example 3, Comparative Example 2, and Comparative Example 1, respectively.

In the mung bean leaves of Example 1, daidzin was 11,302.36 μg/g, genistin was 1944.66 μg/g, malonyl daidzin was 110.86 μg/g, malonyl genistin was 214.12 μg/g, and daidzein was 668.99 μg/g. g and genistein were significantly increased to 92.24 ㎍/g.

Test Example 5: Comparative analysis with ethylene treated soybean leaves

Soybean plants were grown using soybean seeds in the same manner as in Preparation Example 1, treated with ethylene, harvested soybean leaves, and analyzed for beta-glycoside isoflavone derivative content in the same manner as in Test Example 4. Example of Test Example 4 The content of beta-glycoside isoflavone derivatives in the mung bean leaves of 1 was compared, and the results are shown in Figure 11.

As shown in Figure 11, it can be seen that the daidzin and genistin contents of mung bean leaves grown according to the present invention (Example 1) are 7.5 and 1.45 times higher, respectively, than those of soybean leaves grown according to the present invention. there is.

Preparation Example 2: Preparation of pharmaceutical composition

<1-1> Production of powder

Mung bean leaves enhanced with isoflavone derivatives of the present invention, 0.1 g of extract or fraction of extract thereof, 1.5 g of lactose and 0.5 g of talc.

The above ingredients were mixed and filled into an airtight bubble to prepare a powder.

<1-2> Preparation of tablets

0.1 g of mung bean leaves enhanced with the isoflavone derivative of the present invention, extract or fraction of the extract, 7.9 g of lactose. 1.5 g crystalline cellulose and 0.5 g magnesium stearate

After mixing the above ingredients, tablets were prepared by direct tableting method.

<1-3> Manufacturing of capsules

Mung bean leaves enhanced with isoflavone derivatives of the present invention, 0.1 g of extract or fraction of extract thereof, 5 g of corn starch and 4.9 g of carboxycellulose

After mixing the above ingredients to prepare a powder, the powder was filled into a hard capsule according to a typical capsule manufacturing method to prepare a capsule.

<1-4> Preparation of injections

Mung bean leaves enhanced with the isoflavone derivative of the present invention, 0.1 g of extract or fraction of the extract, appropriate amount of sterilized distilled water for injection, appropriate amount of pH adjuster

It was prepared with the above ingredients per 1 ampoule (2 ml) according to the usual manufacturing method for injections.

Manufacturing Example 3: Manufacturing of health functional food

<1-1> Manufacturing of mixed tea

A mixed tea was prepared by mixing mung bean leaves with enhanced isoflavone derivatives of the present invention, extracts or fractions of extracts thereof, and herbal plants with flavor.

<1-2> Manufacturing of flour food

0.5 to 5.0 parts by weight of mung bean leaves, extracts or extract fractions enhanced with the isoflavone derivative of the present invention were added to flour, and the mixture was used to prepare bread, cakes, cookies, crackers and noodles.

<1-3> Preparation of soups and gravies

0.1 to 5.0 parts by weight of mung bean leaves, extracts or fractions of the extracts enhanced with the isoflavone derivative of the present invention were added to soups and gravy to prepare health-enhancing meat products and noodle soup and gravy.

<1-4> Production of ground beef

Ground beef for health promotion was prepared by adding 10 parts by weight of mung bean leaves, extracts or fractions of the extracts enhanced with the isoflavone derivative of the present invention to ground beef.

<1-5> Manufacturing of dairy products

5 to 10 parts by weight of mung bean leaves, extracts or fractions of the extracts enhanced with the isoflavone derivative of the present invention were added to milk, and various dairy products such as butter and ice cream were manufactured using the milk.

<1-6> Manufacturing of Seonsik

Brown rice, barley, glutinous rice, and coix radish were gelatinized using a known method, dried, roasted, and then made into powder with a particle size of 60 mesh using a grinder.

Black beans, black sesame seeds, and perilla seeds were steamed and dried using a known method, roasted, and then made into powder with a particle size of 60 mesh using a grinder.

Mung bean leaves, extracts or fractions of the extracts enhanced with the isoflavone derivative of the present invention were concentrated under reduced pressure in a vacuum concentrator, dried with a spray and hot air dryer, and the dried product was pulverized with a grinder to a particle size of 60 mesh to obtain a dry powder.

It was prepared by mixing the grains and seeds prepared above, mung bean leaves enhanced with the isoflavone derivative of the present invention, and extracts or fractions of the extracts in the following ratio.

Cereals (30 parts by weight brown rice, 15 parts by weight, barley 20 parts by weight), seeds (7 parts by weight perilla seeds, 8 parts by weight black beans, 7 parts by weight black sesame seeds), soybean leaves fortified with the isoflavone derivative of the present invention, soybeans Stems and bean roots, extracts or fractions of extracts (3 parts by weight), Reishi (0.5 parts by weight), Rehmannia glutinosa (0.5 parts by weight)

<1-7> Manufacturing of beverages

Mung bean leaves enhanced with the isoflavone derivative of the present invention, 100 mg of extract or extract fraction thereof, 100 mg of citric acid, 100 mg of oligosaccharide, 2 mg of plum concentrate, 100 mg of taurine, and purified water were added to make a total of 500 ml.

After mixing the above ingredients according to a typical health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered, obtained in a sterilized 1 L container, sealed, sterilized, and refrigerated to prepare the present invention. It is used in the production of health drink compositions.

The composition ratio is a mixture of ingredients relatively suitable for beverages of preference in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as demand class, demand country, and use purpose.

Manufacturing Example 4: Cosmetic manufacturing

<1-1> Cream production

Mung bean leaves enhanced with the isoflavone derivative of the present invention, 4.6 parts by weight of extract or fraction of the extract, 2.8 parts by weight of cetostearyl alcohol, 2.6 parts by weight of beeswax, 1.4 parts by weight of stearic acid, 2 parts by weight of lipophilic glycerin monostearate, blood Easy-100 stearate 1 part by weight, sorbital sesquioleate 1.4 parts by weight, jojoba oil 4 parts by weight, squalane 3.8 parts by weight, polysorbate 60 1.1 parts by weight, macadamia oil 2 parts by weight, tocopherol acetate 0.2 parts by weight, methyl 0.4 parts by weight of polysiloxane, 0.1 part by weight of ethylparaben, 0.1 part by weight of propylparaben, 0.1 part by weight of Euxyl K-400, 7 parts by weight of 1,3-butylene glycol, 0.05 parts by weight of methylparaben, 6 parts by weight of glycerin, d-panthenol It was prepared according to the cream manufacturing method by mixing 0.2 parts by weight of triethanolamine, 0.2 parts by weight of pt 41891, and 0.295 parts by weight of p-H2O 46.05.

<1-2> Production of lotion

3.5 parts by weight of the extract or fraction of the extract of mung bean leaves enhanced with the isoflavone derivative of the present invention, 1.6 parts by weight of cetostearyl alcohol, 1.4 parts by weight of stearic acid, 1.8 parts by weight of lipophilic glycerin monostearate, 2.6 parts by weight of PEG-100 stearate. Parts by weight, sorbital sesquioleate 0.6 parts by weight, squalene 4.8 parts by weight, macadamia oil 2 parts by weight, jojoba oil 2 parts by weight, tocopherol acetate 0.4 parts by weight, methylpolysiloxane 0.2 parts by weight, ethylparaben 0.1 parts by weight, propylparaben 0.1 Parts by weight, 1,3-butylene glycol 4 parts by weight, 0.1 parts by weight of methylparaben, 0.1 parts by weight of xanthan gum, 4 parts by weight of glycerin, 0.15 parts by weight of d-panthenol, 0.1 parts by weight of allantoin, carboxylic acid (2% aq. It was prepared according to the lotion manufacturing method by mixing 4 parts by weight of Sol), 0.15 parts by weight of triethanolamine, 3 parts by weight of ethanol, 0.1 parts by weight of pt 41891, and 48.3 parts by weight of p-H20.

1: Light source control device (red, blue, white light source)
2: Light source control device (UV-A or UV-B)
3: LED light source (red/blue/white/UV-A or B)
4: Automatic temperature and humidity control and data collection device
5: Temperature, humidity and carbon dioxide meter
6: Ventilation fan
7: Temperature control fan
8: Water circulation device for temperature control
9: Ethylene gas cylinder
10: Ethylene gas injection device
11: Ethylene gas meter
12: Data storage device
13: Plant growth observation window

Claims (16)

A method of cultivating mung bean leaves with enhanced isoflavone derivatives of the following formula (1):
[Formula 1]
wherein R ₁ is H or OH, R ₂ is glucose or glucose malonate,
The above method is
i) sowing mung bean seeds or germinated mung beans in a cultivation facility and growing them for 20 to 40 days;
ii) treating the grown mung beans with ethylene at a concentration of 2,500 to 10,000 ppm 2 to 3 times for 8 to 12 hours each; and
iii) harvesting mung bean leaves.
The method of claim 1, wherein the cultivation facility is selected from the group consisting of smart farms, plant factories, plant cultivation devices, vertical farms, vertical cultivation devices, container cultivation devices, hydroponic cultivation devices, smart chambers, greenhouse systems, and cultivation systems. A method of cultivating mung bean leaves with enhanced isoflavone derivatives.
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 2, wherein the cultivation facility is a vertical plant factory or a smart chamber.
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 1, wherein germinated mung beans are used for sowing in step i).
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 1, wherein in step i), the mung bean leaves are grown for 30 to 35 days after sowing.
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 1, wherein the ethylene concentration in step ii) is 5,000 to 1,0000 ppm.
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 1, wherein in step ii), ethylene at a concentration of 10,000 ppm is treated three times for 8 hours each.
The method of cultivating mung bean leaves with enhanced isoflavone derivatives according to claim 1, wherein in step ii), ethylene at a concentration of 10,000 ppm is treated twice for 12 hours each.
Mung bean leaves with enhanced isoflavone derivatives prepared by the cultivation method according to claim 1.
The mung bean leaves with enhanced isoflavone derivatives according to claim 9, wherein the mung bean leaves contain more than 14,333 μg/g of isoflavone derivatives.
The method of claim 9, wherein the mung bean leaves contain more than 13,347 μg/g of beta-glycoside isoflavone derivatives, more than 324 μg/g of maloyl-beta-glycoside isoflavone derivatives, and aglycone isoflavones. Mung bean leaves enhanced with isoflavone derivatives, containing more than 761 μg/g of the derivative.
The mung bean leaf with enhanced isoflavone derivatives according to claim 9, wherein the isoflavone derivative is at least one selected from the group consisting of Formulas 2 to 6 below:

A health functional food containing mung bean leaves with enhanced isoflavone derivatives according to claim 9.
A cosmetic comprising mung bean leaves enhanced with isoflavone derivatives according to claim 9.
A pharmaceutical composition comprising mung bean leaves enhanced with isoflavone derivatives according to claim 9,
The pharmaceutical composition is for preventing and treating diseases caused by estrogen deficiency.
A health functional food for estrogen supplementation containing as an active ingredient mung bean leaves enhanced with the isoflavone derivative according to claim 9.