KR20240108834A - Method for cultivation of mung bean leaves with increased isoflavone derivatives and its use - Google Patents
Method for cultivation of mung bean leaves with increased isoflavone derivatives and its use Download PDFInfo
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- KR20240108834A KR20240108834A KR1020220190325A KR20220190325A KR20240108834A KR 20240108834 A KR20240108834 A KR 20240108834A KR 1020220190325 A KR1020220190325 A KR 1020220190325A KR 20220190325 A KR20220190325 A KR 20220190325A KR 20240108834 A KR20240108834 A KR 20240108834A
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- South Korea
- Prior art keywords
- mung bean
- bean leaves
- isoflavone derivatives
- enhanced
- isoflavone
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- A—HUMAN NECESSITIES
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/40—Fabaceae, e.g. beans or peas
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- A—HUMAN NECESSITIES
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A—HUMAN NECESSITIES
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- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
본 발명에서는 이소플라본 유도체가 증진된 녹두 잎의 재배방법 및 그 이용이 개시된다.
본 발명에 따른 녹두 잎의 재배방법에 의하면, 단기간 (20~40일)에 이소플라본 유도체가 현저히 증진된 녹두 잎을 재배할 수 있다. 또한 본 발명의 재배방법은 이소플라본 유도체 고 함량 녹두 잎을 연중 안정적으로 생산할 수 있다.
본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎, 그 추출물 또는 그 분획물은 에스트로겐 대체재로 사용될 수 있고, 에스트로겐 결핍에 기인한 질환의 예방 및 개선용, 항산화용, 및 피부미용용 약제학적 조성물, 간강기능성 식품 및 화장품의 소재로 유용하다.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
본 발명은 이소플라본 유도체가 증진된 녹두 잎의 재배방법 및 그 이용에 관한 것으로, 더 상세하게는 특정 조건의 에틸렌 처리를 이용하여 이소플라본 유도체가 증진된 녹두 잎의 재배방법 및 이소플라본 유도체가 증진된 녹두 잎의 이용에 관한 것이다.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.
여성호르몬인 에스트로겐은 40대 이후 줄어들기 시작하여 궁극적으로 갱년기를 유발한다. 에스트로겐이 줄어들면 이 호르몬의 작용점인 뼈, 심장, 피부, 뇌 등에 노화가 시작된다. 이러한 노화 진행을 억제하기 위해 에스트로겐 대체제가 절실히 요구되며 구조적인 유사성을 갖는 이소플라본 유도체가 효과적인 에스트로겐 대체제로 보고되고 있다.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.
이소플라본 유도체는 약리학적 효능이 뛰어난 대표적인 플라보노이드로서 전세계적으로 이들이 고-함유된 기능성 식물개발에 많은 자본이 투자되고 있는 실정이다. 이소플라본은 비스테로이드 에스트로겐으로 화학적 및 생리학적 특성이 에스트로겐과 유사하여 식물성 에스트로겐이라 불리며 경쟁적으로 에스트로겐 수용체와 결합함으로써 에스트로겐 또는 항 에스트로겐 작용을 하는 물질이다(Hudson, 2001). 이들은 여성호르몬인 에스트로겐과 구조가 유사하여 에스트로겐에서 나타나는 유용한 생리활성을 보여준다. 이소플라본을 섭취하면, 여성의 내분비 상태를 조절하고 난소의 주기성에 영향을 미친다고 알려져 있다(Kenneth, Setchell, & Cassidy, 1999). 특히 여성호르몬이 부족하여 나타날 수 있는 갱년기 증세 중 하나인 골다공증의 위험을 감소시키고, 혈장 내 콜레스테롤 수치를 낮추는 것으로 보고되고 있다(Obstetrics & Gynecology 2001. 389). 또한 관상동맥심장질환(coronary arterial heart disease, CAHD)의 위험을 낮추는 효과와 항산화 활성이 뛰어난 것으로 알려져 있다. 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.
이소플라본 유도체 중 비배당체로는 제니스테인(genistein), 다이드제인(daidzein) 및 글리시테인(glycitein) 등이 있으며, 배당체로는 제니스틴(genistin), 다이드진(daidzin) 및 글리시틴(glycitin) 등이 있다. 대두 유래 비 발효식품에는 배당체가 많은 반면, 된장과 같은 대두를 이용한 발효식품에는 발효에 관계된 박테리아에 의해 배당체로부터 당이 분해되면서 비배당체가 주로 존재하게 된다. 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.
콩과 작물(콩, 녹두 등)의 이소플라본 유도체는 에스트로겐 수용체인 ER-베타(estrogen receptor beta)에 높은 친화도를 가지고 있어 가장 효과적인 에스트로겐 대체제이다(Endocrinology 1997, 863). 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).
이와 같이 녹두 이소플라본 유도체는 에스트로겐 대체재로 유용한 생물소재이지만, 녹두에 포함된 단백질, 지방 및 탄수화물의 함량이 높아 이소플라본 유도체의 추출, 정제 및 제품화와 같은 가공과정에 어려움이 있다. 또한 녹두 잎은, 콩잎과 다르게, 식품 등으로 이용되지 못하고 버려지고 있는 실정이다. 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.
이소플라본 유도체가 고 함유된 작물 연구는 주로 콩을 대상으로 진행되고 있으며, 최근에 콩잎에 에틸렌 또는 에틸렌 공여체인 에테폰을 처리하여 이소플라본 유도체 고함유 콩잎을 생산하는 재배 방법 기술이 보고되었다 (한국등록특허 제10-1451298, 한국등록특허 제10-2282625). 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).
따라서 버려지는 녹두 잎을 이용할 수 있도록 이소플라본 유도체 고 함량 녹두 잎의 재배방법의 개발이 요구되고 있다. 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.
이에, 본 발명자들은 종래 기술에서의 요구에 부응하기 위해 지속적으로 연구한 결과, 특정 조건의 에틸렌 처리를 통하여 이소플라본 유도체가 현저히 증진된 녹두 잎을 재배할 수 있음을 확인하고 본 발명을 완성하였다.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.
본 발명의 목적은 이소플라본 유도체가 증진된 녹두 잎의 재배방법을 제공하는 것이다.The purpose of the present invention is to provide a method for cultivating mung bean leaves with enhanced isoflavone derivatives.
본 발명의 또 다른 목적은 상기 방법에 의해 재배된 이소플라본 유도체가 증진된 녹두 잎을 제공하는 것이다.Another object of the present invention is to provide mung bean leaves with enhanced isoflavone derivatives grown by the above method.
본 발명의 또 다른 목적은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 건강기능성 식품을 제공하는 것이다.Another object of the present invention is to provide a health functional food containing mung bean leaves with enhanced isoflavone derivatives.
본 발명의 또 다른 목적은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 화장품을 제공하는 것이다.Another object of the present invention is to provide cosmetics containing mung bean leaves with enhanced isoflavone derivatives.
본 발명의 또 다른 목적은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 약제학적 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition comprising mung bean leaves enhanced with the isoflavone derivatives.
본 발명의 목적에 따라서, 본 발명은 하기 화학식 1의 이소플라본 유도체가 증진된 녹두 잎의 재배방법을 제공한다: 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):
[화학식 1] [Formula 1]
상기 식에서, R1은 H 또는 OH이고, R2는 글루코오스(glucose) 또는 글루코오스 말로네이트(glucose malonate)이다. In the above formula, R 1 is H or OH, and R 2 is glucose or glucose malonate.
상기 재배방법은 The cultivation method is
i) 재배 시설에서 녹두 종자 또는 발아된 녹두를 파종하여 20 ~ 40일을 생육시키는 단계; i) sowing mung bean seeds or germinated mung beans in a cultivation facility and growing them for 20 to 40 days;
ii) 생육된 녹두에 2,500 ~ 10,000 ppm의 농도의 에틸렌을 8 ~ 12 시간씩 2 ~ 3회 처리하는 단계; 및 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) 녹두 잎을 수확하는 단계;를 포함한다. iii) harvesting mung bean leaves.
본 발명에서 화학식 1의 이소플라본 유도체는 하기 [화학식 2] 내지 [화학식 7]로 표기되는 화합물 중 어느 하나 이상 일 수 있다: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]:
단계 i) 생육Step i) Growth
재배 시설에서 녹두 종자 또는 발아된 녹두를 파종하여 20 ~ 40일을 생육시킨다.Mung bean seeds or germinated mung beans are sown in a cultivation facility and grown for 20 to 40 days.
본 발명에서 '재배 시설'은 인공적으로 증식환경을 조절할 수 있는 재배 시설이면 제한되지 않으며, 스마트팜, 식물공장, 식물재배장치, 수직농장, 수직재배장치, 컨테이너 재배장치, 수경 재배장치, 스마트 챔버, 온실시스템, 재배시스템 등일 수 있으며, 바람직하게는 수직형 식물공장 또는 스마트 챔버일 수 있다. 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.
일례로 수직형 식물공장은 내부공간을 포함하는 식물공장부; 상기 식물공장부 중앙에 설치된 도어부; LED 조명 시설, 온도조절을 위한 냉·난방 시설, 식물공장부 환기를 위한 환기 시설; 관수 시설; 원격으로 자동화 시스템을 위한 ICT 설비; 및 운영 어플을 포함할 수 있다 (도 1a 및 도 1b).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).
또 다른 일례로 스마트 챔버는 내부공간을 포함하는 챔버부; 상기 챔버부를 개폐하는 도어부; 상기 도어부 중앙에 설치된 관찰 창; 챔버부 내부 천장에 설치된 LED 광원부; 온도, 습도 및 이산화탄소 측정기; 챔버부 환기를 위한 환기 팬; 챔버부 내부 뒷면에 설치된 온도조절 팬; 에틸렌 가스 분사장치; 에틸렌 가스 측정기; LED 광원을 조절하는 광원조절장치; 온도 및 습도 자동제어 및 데이터 수집장치; 온도조절용 물 순환장치; 에틸렌 가스통; 및 데이터 저장장치를 포함할 수 있다 (도 2a 및 도 2b). 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).
단계 i)에서, 파종은 녹두 종자 또는 발아된 녹두를 이용할 수 있으며, 바람직하게는 발아된 녹두를 파종한다. In step i), mung bean seeds or germinated mung beans can be used for sowing, and preferably sprouted mung beans are sowed.
본 발명에서 발아된 녹두는 녹두종자 무게에 1~3배 정도의 물을 넣고 15~25℃에서 녹두를 불린 후 20~30℃의 발아기 또는 콩나물 제조기에 넣고 암실에서 16~24시간 내 발아를 시키는 것으로 제조될 수 있다.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
단계 i)에서 녹두의 생육은 파종 후 20~40 일 수행되며, 바람직하게는 30~35 일이 바람직하다. 20일 이하의 경우 녹두의 바이오매스 함량이 적고 40일 이상의 경우 재배시설에서 생육에 문제가 발생한다.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.
녹두 생육을 위하여 재배 시설에 설치된 LED 광원부는 청색과 적색의 이중 혼합광원과 백색의 식물성장 LED를 혼합하여 사용한다.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.
녹두 생육을 위한 재배 시설의 광량(Lux)은 약 10,000 ~15,000를 사용한다. 10,000 Lux 미만의 경우 광합성이 원활하지 않아 녹두 바이오매스(잎) 함량이 적고 이소플라본 유도체의 생성이 원활지 않고, 15,000 Lux 초과일 경우 센 광량에 의하여 생육이 원활하지 않고 잎 고사하는 경우가 발생한다.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. .
녹두 생육을 위한 재배 시설 내의 온도는 20 ~ 35℃인 것이 바림직하고, 25~30℃인 것이 더욱 바람직하다. 20℃ 미만의 경우 냉해로 인해 녹두가 생육하지 못하고 35℃ 초과의 경우 고온 장애가 발생할 수 있다.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.
녹두 생육을 위한 재배 시설 내의 습도는 40 ~ 80%인 것이 바람직하고, 50 ~ 60%인 것이 더욱 바람직하다. 40% 미만의 경우 수분 부족으로 녹두의 광합성 등은 신진대사가 원활하지 않아 잎 등이 고사할 수 있고 80% 초과의 경우 과다한 수분으로 뿌리 등이 썩을 수 있다.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.
단계 ii) 에틸렌 처리Step ii) Ethylene treatment
생육된 녹두에 2,500 ~ 10,000 ppm의 농도의 에틸렌을 8 ~ 12 시간씩 2 ~ 3회 처리한다.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.
에틸렌 처리 농도는 2,500 ~ 10,000 ppm의 고농도이며, 바람직하게는 5,000~1,0000 ppm의 농도로 처리한다. 에틸렌 처리 농도가 2,500 ppm 미만의 경우 녹두 잎에 이소플라본 유도체 축적이 거의 이루어지 않고 10,000 ppm 초과 처리해도 이소플라본 유도체 축적에 큰 차이가 없다.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.
1회 에틸렌 처리시간은 8~12시간이 바람직하다. 에틸렌 처리시간이 8시간 미만의 경우 이소플라본 유도체 축적이 충분히 이루어지 않고 12시간 초과의 경우에도 이소플라본 유도체 축적에 큰 차이가 없다.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.
에틸렌 처리는 2~3회 처리하는 것이 바람직하다. 에틸렌 처리가 2회 미만의 경우 이소플라본 유도체 축적이 충분히 이루어지 않고 3회 초과의 경우에는 이소플라본 유도체 축적에 큰 차이가 없다.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.
단계 ii)에서 에틸렌 처리는 재배시설을 완전히 닫힌 밀폐된 공간에서 이루어진다. 즉 재배시설의 도어를 닫고 환기팬을 막아서 밀폐를 시킨 후 에틸렌을 처리한다. 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.
에틸렌 처리와 다음 에틸렌 처리 사이 시간은 1 ~ 2시간 정도로 재배시설을 오픈하여 환기시켜 에틸렌을 제거하여 녹두를 안정화시킨다. 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.
에틸렌 처리 전에 상기와 같은 밀폐된 공간에서 녹두를 1 ~ 2시간 유지하여 안정화시킨다. 녹두의 호흡에 의해서 발생하는 수분으로 인해 습도조절을 위해 안정화시킨다.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.
에틸렌 처리시, 재배시설은 백색, 청색, 적색의 단일광원; 백색과 청색, 백색과 적색, 청색과 적색의 이중 혼합광원; 또는 백색과 청색과 적색의 삼중 혼합광원이 사용될 수 있으며, 백색과 청색과 적색의 삼중 혼합광원이 바람직하며, 광량은 100 ㎛ol/m2/s 이상을 사용하며, 바람직하게는 200 ㎛ol/m2/s 이상을 사용한다.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.
에틸렌 처리시, 재배시설의 온도는 20~40℃인 것이 바람직하고, 30~35℃인 것이 더욱 바람직하다. 습도는 60~90%인 것이 바람직하고, 70~80%인 것이 더 바람직하다. 에틸렌 처리 온도 20℃ 및 습도 60% 미만의 경우 녹두의 대사작용이 원활하지 않아 이소플라본 유도체 축적이 거의 이루어지지 않고 온도 40℃ 및 습도 90% 초과 역시 고온과 과다 수분 함량으로 인해서 호흡 등의 대사작용이 원활하지 않아 이소플라본 유도체 이루어지지 않는다.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) 수확iii) Harvest
단계 ii)에서 에틸렌 처리가 완료된 녹두 잎을 수확한다. Harvest the mung bean leaves on which ethylene treatment has been completed in step ii).
상기와 같이 본 발명에 따른 재배방법은 녹두를 수확 전에 단계 ii)와 같이 특정 조건으로 에틸렌을 처리함에 따라서 녹두 잎의 이소플라본 유도체의 함량을 현저히 증진시킬 수 있다. 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.
본 발명에 따라 재배된 녹두 잎의 경우 (약 14,333 ㎍/g; 실시예 1), 수직형 식물공장에서 생육되고 에틸렌이 처리되지 않은 녹두 잎 (약 1,016 ㎍/g; 비교예 3), 발아 녹두 (약 48 ㎍/g; 비교예 2) 및 녹두 종자 (약 1.8 ㎍/g; 비교예 1)에 비하여, 이소플라본 유도체의 함량이 각각 약 14.1배, 약 298.63배 및 약 7,962 배로 현저히 증진되었다 (도 10).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).
본 발명의 또 다른 목적에 따라서, 상기 방법에 의해 재배된 이소플라본 유도체가 증진된 녹두 잎을 제공한다.According to another object of the present invention, there is provided mung bean leaves enhanced with isoflavone derivatives grown by the above method.
본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎은 이소플라본 유도체를 14,333 ㎍/g 이상 함유하고, 베타-글리코시드 이소플라본 유도체를 13,347 ㎍/g 이상 함유하고, 말로일-베타-글로코시드 이소플라본 유도체를 324 ㎍/g 이상 함유하고, 아글리콘 이소플라본 유도체를 761 ㎍/g 이상 함유한다 (표 7, 도 10).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).
본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎은 그 자체로, 또는 용매로 추출되어 식품, 화장품 및 약제학적 조성물의 소재로 사용될 수 있다.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.
본 발명에서 추출 용매로는 물, 알코올 또는 이들의 혼합물을 사용할 수 있다. 알코올로는 C1 내지 C2 저급 알코올을 이용하는 것이 바람직하며, 저급 알코올로는 에탄올 또는 메탄올을 이용하는 것이 바람직하다. 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.
추출 방법으로는 진탕추출, Soxhlet 추출 또는 환류 추출을 이용하는 것이 바람직하나 이에 한정되지 않는다. 구체적으로는 추출 용매를 건조된 녹두 잎에 1~10배(v/w) 첨가하여 추출하는 것이 바람직하고, 추출온도는 20~100℃인 것이 바람직하고, 추출시간은 10~48시간인 것이 바람직하나 이에 한정되지는 않는다. 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.
추출물은 감압 농축한 후 건조될 수 있으며, 감압농축은 진공감압농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나 이에 한정하지 않는다. 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.
추출물은 추가로 유기용매로 분획될 수 있으며, 출물을 물에 현탁시킨 후 유기용매로 n-헥산, 클로로포름, 에틸아세테이트 또는 부탄올이 사용하여 분획 할 수 있으며, 클로로포름을 사용하는 것이 바람직하다. 추출물로부터 분획 과정을 1~5회, 바람직하게는 3회 반복하여 분획물을 수득할 수 있고, 분획 후 감압 농축하는 것이 바람직하나 이에 한정하지 않는다.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.
본 발명의 또 다른 목적에 따라서, 본 발명은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 건강기능성 식품을 제공한다.According to another object of the present invention, the present invention provides a health functional food containing mung bean leaves with enhanced isoflavone derivatives.
본 발명에 따른 건강기능성 식품은 에스트로겐 결핍에 기인한 질환의 예방 및 개선용, 항산화용, 및 피부미용용으로 유용하다. 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.
본 발명에서 상기 에스트로겐 결핍에 기인한 질환은 골다공증, 심장질환, 유방암, 외음질 질환, 과지질혈증, 피부노화 및 안면홍조로 구성된 군으로부터 선택된 어느 하나이다. 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.
이소플라본 유도체들은 상기와 같은 에스트로겐 결핍에 기인한 질환의 예방 및 개선용, 항산화용, 및 피부미용용으로 생리활성이 확립되어 있으므로, 고함량의 이소플라본 유도체를 포함하는 본 발명에 따른 녹두 잎은 상기와 같은 건강기능성 식품으로 유용하게 사용될 수 있다.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.
본 발명에 따른 건강기능성 식품은 이소플라본 유도체가 증진된 녹두 잎을 그대로 포함하거나, 상기와 같은 추출물 또는 분획물의 형태로 다른 식품 또는 식품 성분과 함께 포함될 수 있다. 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.
본 발명에서 '건강기능성 식품'이란 일상 식사에서 결핍되기 쉬운 영양소나 인체에 유용한 기능을 가진 원료나 성분을 사용하여 제조한 식품으로, 인체의 건강을 유지하는데 도움을 주는 식품을 의미한다. 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.
본 발명에서 건강기능성 식품의 형태 및 종류는 특별히 제한되지 않는다. 구체적으로는 정제, 캅셀, 분말, 과립, 액상 및 환의 형태일 수 있다. 건강기능성 식품은 추가성분으로서 여러 가지 향미제, 감미제 또는 천연 탄수화물을 포함할 수 있다. 감미제는 천연 또는 합성 감미제일 수 있고, 천연 감미제의 예로는 타우마틴, 스테비아 추출물 등이 있다. 한편, 합성 감미제의 예로는 사카린, 아스파탐 등이 있다. 또한 천연 탄수화물은 모노사카라이드, 디사카라이드, 폴리사카라이드, 올리고당 및 당알코올 등일 수 있다.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.
본 발명에서 건강기능성 식품은 상기 추가성분 외에, 영양제, 비타민, 전해질, 풍미제, 착색제, 펙스탄 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 등을 더 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합으로 사용될 수 있다. 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.
본 발명에서 상기 식품의 종류는 특별히 제한되지 않으며, 환제, 정제, 캡슐제, 차, 선식, 밀가루 식품, 스프, 유제품, 소스, 육류 첨가제, 음료 (파우치제, 드링크제 등) 등일 수 있으나, 이에 제한되지는 않는다. 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.
바람직하게는 본 발명에 따른 식품은 차(Tea)일 수 있다. 본 발명에 따른 차는 본 발명에 따른 이소플라본 유도체가 강화된 녹두 잎 및 향미를 갖는 허브 식물체를 혼합한 혼합차일 수 있다. 상기 허브 식물체는 캐모마일, 레몬그라스, 로즈힙, 라벤다, 페퍼민트, 펜넬, 로즈마리, 자스민, 히비스커스, 로즈플라워, 애플후르츠, 스트로베리후르츠, 레몬후르츠 및 오렌지 플라워로 구성된 군으로부터 선택된 하나 이상인 것이 바람직하다. 본 발명의 차 또는 혼합차는 착향차(着香茶), 티백(tea bag), 인스턴트(instant tea) 및 캔(can)으로 구성된 군으로부터 선택된 형태로 가공될 수 있다.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.
본 발명의 또 다른 목적에 따라서, 본 발명은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 화장품을 제공한다.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.
본 발명에 따른 화장품은 이소플라본 유도체가 증진된 녹두 잎을 그대로 포함하거나, 상기와 같은 추출물 또는 분획물의 형태로 다른 화장품 성분과 함께 포함될 수 있다. 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.
본 발명에서 화장품은 로션, 연고, 겔, 크림, 패치, 마스크팩 또는 분무제 등의 형태일 수 있으나 여기에 국한되는 것은 아니며, 추가로 지방 물질, 유기 용매, 용해제, 농축제 및 겔화제, 연화제, 항산화제, 현탁화제, 안정화제, 발포제, 방향제, 계면활성제, 물, 이온형 또는 비이온형 유화제, 충전제, 금속이온봉쇄제 및 킬레이트화제, 보존제, 비타민, 차단제, 습윤화제, 필수 오일, 염료, 안료, 친수성 또는 친유성 활성제, 지질 소낭 등 화장품에 통상적으로 사용되는 임의의 첨가제를 함유할 수 있다.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.
본 발명의 또 다른 목적에 따라서, 본 발명은 상기 이소플라본 유도체가 증진된 녹두 잎을 포함하는 에스트로겐 결핍에 기인한 질환의 예방 및 치료용 약제학적 조성물을 제공한다. 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.
본 발명에서 상기 에스트로겐 결핍에 기인한 질환은 골다공증, 심장질환, 유방암, 외음질 질환, 과지질혈증, 피부노화 및 안면홍조로 구성된 군으로부터 선택된 어느 하나이다. 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.
이소플라본 유도체들은 에스트로겐 대체재로서 상기와 같은 에스트로겐 결핍에 기인한 질환의 예방 및 개선용으로 확립되어 있으므로, 고함량의 이소플라본 유도체를 포함하는 본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎은 상기와 같은 용도의 약제학적 조성물로 유용하다.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.
본 발명에 따른 약제학적 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화될 수 있다. 상기 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토오스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 혼합생약재에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.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.
본 발명에 따른 약제학적 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여시 피부 외용 또는 복강내주사, 직장 내주사, 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식으로 투여될 수 있다.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.
본 발명에 따른 약제학적 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 바람직한 효과를 위해서, 상기 조성물은 1일 0.0001 내지 1 g/kg으로, 바람직하게는 0.001 내지 200 mg/kg으로 투여하는 것이 바람직하다. 상기 투여는 하루에 한 번 투여할 수도 있고, 수 회 나누어 투여할 수도 있다.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.
본 발명에 따른 녹두 잎의 재배방법에 의하면, 단기간 (20~40일)에 이소플라본 유도체가 현저히 증진된 녹두 잎을 재배할 수 있다. 또한 본 발명의 재배방법은 이소플라본 유도체 고 함량 녹두 잎을 연중 안정적으로 생산할 수 있다. 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.
본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎, 그 추출물 또는 그 분획물은 에스트로겐 대체재로 사용될 수 있다. Mung bean leaves, extracts thereof, or fractions thereof enhanced with isoflavone derivatives according to the present invention can be used as an estrogen substitute.
본 발명에 따른 이소플라본 유도체가 증진된 녹두 잎, 그 추출물 또는 그 분획물은 에스트로겐 결핍에 기인한 질환의 예방 및 개선용, 항산화용, 및 피부미용용 약제학적 조성물, 간강기능성 식품 및 화장품의 소재로 유용하다. 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.
도 1a는 본 발명에 따른 재배 시설의 일종인 수직형 식물공장의 구성도의 일례를 도시한 것이다. 도 1b는 수직형 식물공장의 내부 시설 및 자동제어시스템의 일례를 도시한 것이다. 도 1c는 수직형 식물공장에서 파종 후 15일 및 35일 생육시 녹두의 모습이다.
도 2a는 본 발명에 따른 재배 시설의 일종인 스마트 대사체 챔버의 구성도의 일례를 도시한 것이다. 도 2b는 스마트 대사체 챔버의 실제 제작된 제품의 보여주는 사진이다. 도 2c는 에틸렌 처리 전/후 녹두의 모습이다.
도 3a은 녹두 종자의 이소플라본 유도체를 분석한 HPLC 크로마토그램이며 (비교예 1), 도 3b는 발아 녹두의 이소플라본 유도체를 분석한 HPLC 크로마토그램이며 (비교예 2), 도 3c는 수직형 식물공장에서 파종 후 35일 생육된 녹두 잎의 이소플라본 유도체를 분석한 HPLC 크로마토그램이며 (비교예 3), 도 3d는 수직형 식물공장에서 파종 후 35일 생육시킨 후 스마트 대사체 챔버에서 에틸렌 처리한 녹두 잎의 이소플라본 유도체를 분석한 HPLC 크로마토그램이다 (실시예 1).
a는 다이드진, b는 제니스틴, c는 말로닐 다이드진, d는 말로닐 제니스틴, e는 다이드제인, f는 제니스테인.
도 4은 도 3d HPLC 크로마토그램에서 피크 a의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 다이드진의 화학식을 나타낸 것이다.
도 5은 도 3d HPLC 크로마토그램에서 피크 b의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 제니스틴의 화학식을 나타낸 것이다.
도 6은 도 3d HPLC 크로마토그램에서 피크 c의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 말로닐 다이드진의 화학식을 나타낸 것이다.
도 7은 도 3d HPLC 크로마토그램에서 피크 d의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 말로닐 제니스틴의 화학식을 나타낸 것이다.
도 8은 도 3d HPLC 크로마토그램에서 피크 e의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 다이드제인의 화학식을 나타낸 것이다.
도 9은 도 3d HPLC 크로마토그램에서 피크 f의 LC-ESI-TOF/MS의 BPI 스펙트럼 및 해당 화합물인 제니스테인의 화학식을 나타낸 것이다.
도 10는 비교예 1, 비교예 2, 비교예 3 및 실시예 1의 녹두 잎의 이소플라본 유도체 함량을 비교한 그래프이다.
도 11은 본 발명에 따른 녹두잎과 에틸렌 처리된 콩잎의 베타-글리코시드 이소플라본 함량을 비교한 그래프이다. 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.
이하, 본 발명을 도면, 실시예 및 제조예에 의해 상세히 설명한다. 단 도면, 실시예 및 제조예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 도면, 실시예 및 제조예에 의해서 한정되는 것은 아니다.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.
<수직형 식물공장><Vertical plant factory>
본 발명에서 녹두 생육에 사용된 수직형 식물공장를 도면을 참조하여 상세히 설명한다.The vertical plant factory used for growing mung beans in the present invention will be described in detail with reference to the drawings.
도 1a는 본 발명에 따른 재배시설의 일종인 수직형 식물공장의 구성도의 일례를 도시한 것이다.Figure 1a shows an example of a configuration diagram of a vertical plant factory, a type of cultivation facility according to the present invention.
도 1a를 참조하면, 본 발명에 따른 수직형 식물공장 외부의 가로×세로× 높이는 3000 mm × 8200 mm × 3000 mm 크기일 수 있으며, 내부의 가로×세로× 높이는 2800 mm × 7700 mm × 2720 mm 크기가 바람직하다. 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.
식물공장의 구성으로 내부공간을 포함하는 식물공장부, 상기 식물공장부 중앙에 설치된 도어부, LED 조명 시설, 온도조절을 위한 냉·난방 시설, 식물공장부 환기를 위한 환기 시설, 관수 시설, 원격으로 자동화 시스템을 위한 ICT 설비 및 운영 어플을 포함한다. 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
상기 수직형 식물공장의 구성으로 LED 조명 시설, 냉·난방 시설, 환기 시설, 관수 시설, ICT 설비, 운영 어플을 포함할 수 있다. 상기 LED 조명 시설은 햇빛과 같은 역할을 하며 일반 LED 조명과 달리 식물 생장용 LED로 재배환경에 맞게 조절 가능하다. 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.
상기 냉·난방 시설은 온도와 습도 조절을 하고 외부 온도 및 습도에 맞춰 축적된 데이터 프로그램을 활용해 온도와 습도 조절이 가능하다.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.
상기 환기 시설은 식물이 숨을 쉴 수 있도록 하는 시설이며, 환기를 통해 산소는 멀리 이동하고 광합성에 필요한 탄산가스가 기공으로 오게 하는 역할한다.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.
상기 관수 시설은 비, 물주기 역할위한 시설로써 자동으로 원하는 시간에 원하는 양만큼 물을 주고 쉽고 간편하게 사용가능하다.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.
상기 ICT 설비는 온도 습도 환기 등을 원격으로 자동제어하는 역할의 시설로 농작업의 시간적/공간적 구속으로부터 자유로워 여유시간이 늘고 삶의 질 개선에 중요한 역할을 한다.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.
상기 운영 어플은 원격 조절 가능 어플리케이션으로 현장에서 직접하지 않고 모바일로 원격 생육환경을 조절하며 빅데이터 활용 작물 생육 정보 분석하는 역할을 한다.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.
<스마트-대사체 챔버><Smart-metabolite chamber>
본 발명에서 에틸렌 처리 단계에서 사용된 스마트 대사체 챔버의 도면을 참조하여 상세히 설명한다.The smart metabolite chamber used in the ethylene treatment step in the present invention will be described in detail with reference to the drawings.
도 2a는 본 발명에 따른 재배시설의 일종인 스마트 대사체 챔버의 구성도의 일례를 도시한 것이다.Figure 2a shows an example of the configuration of a smart metabolite chamber, a type of cultivation facility according to the present invention.
도 2a를 참조하면, 본 발명에 따른 스마트-대사체 챔버는 내부공간을 포함하는 챔버부, 상기 챔버부를 개폐하는 개폐 도어부, 상기 도어부 중앙에 설치된 관찰 창(13), 챔버부 내부 천장에 설치된 LED 광원부(3), 온도, 습도 및 이산화탄소 측정기(5); 챔버부 환기를 위한 환기 팬(6), 챔버부 내부 뒷면에 설치된 온도조절 팬(7), 에틸렌 가스 분사장치(10), 에틸렌 가스 측정기(11), LED 광원을 조절하는 광원조절장치(1, 2), 온도 및 습도 자동제어 및 데이터 수집장치(4), 온도조절용 물 순환장치(8), 에틸렌 가스통(9) 및 데이터 저장장치(12)를 포함할 수 있다. 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).
본 발명에 따른 스마트-대사체 챔버에서 챔버부는 가로×세로× 높이는 500~1200 mm × 400~1000 mm × 400~8000 mm 크기일 수 있으며, 700 mm × 500 mm × 500 mm 크기가 바람직하다. 상기 챔버부의 재질은 알루미늄 또는 스텐레스스틸, 나무, 폴리아크릴판으로 제작하는 바람직하나, 알루미늄 또는 스텐레스스틸로 제작하는 것이 더욱 바람직하다.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.
상기 도어부는 챔버부를 개폐할 수 있게 하고 중앙에 관찰 창(13)이 설치되어 있어 식물체의 생육을 관찰할 수 있다 (도 2c). 관찰 창(13)은 투명 아크릴판, 유리 등의 재질로 구성될 수 있다. 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.
상기 LED 광원부(3)는 챔버부 내부 천장에 설치될 수 있으며 적색, 청색, 백색, UV-A 및 UV-B으로 구성된 군에서 선택되는 1종 이상의 광을 식물체에 조사할 있다. 광량은 암실 조건에서 최대 각각의 LED 광원을 1%에서 100%로 조절될 수 있다. 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.
상기 LED 광원을 조절하는 광원조절장치(1, 2)는 챔버부 외부면의 어느 일부에 설치될 수 있으며, 적색(660 nm), 청색(450 nm) 및 백색의 3채널 광원조절장치(1)와 UV-A(375 nm) 및 UV-B(315 nm)의 광원조절장치(2)로 나뉠 수 있다. 광원조절장치는 각각의 LED 광원의 광량을 1%에서 100%로 조절할 수 있다.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%.
상기 온도, 습도 및 이산화탄소 측정기(5)는 챔버부 내부의 일측면에 설치될 수 있으며 챔버부 외부에 설치된 온도 및 습도 자동제어 및 데이터 수집장치(4)에 연결되어 있다. 온도는 -10℃에서 60℃까지 측정이 가능하며, 습도 측정은 0%에서 100%까지 측정이 가능하다. 이산화탄소는 0에서 2,000 ppm까지 측정이 가능하다The 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.
상기 환기 팬(6)는 챔버부의 환기를 위해 1개 이상 설치될 수 있다. One or more ventilation fans 6 may be installed to ventilate the chamber portion.
상기 온도조절 팬(7)은 챔버부 내부 뒷면에 설치되어 있으며, 챔버부의 내부공간의 온도를 조절하는 기능을 하며 외부에 설치되어 있는 온도조절용 물 순환장치(8)와 연결되어 있다. 스마트-대사체 챔버 내의 온도는 15℃에서 45℃까지 일정하게 유지 가능하며, 스마트-대사체 챔버 내의 실제 습도는 20%에서 100%까지 일정하게 유지 가능하다. 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%.
상기 에틸렌 가스 분사장치(10)는 챔버부 내부의 일측에 설치되어 식물체에 에틸렌을 분사할 수 있으며, 챔버부 외부에 설치된 에틸렌 가스통(9)과 연결되어 있다. 챔버부 내부의 에틸렌 가스 농도는 최대 10,000 ppm까지의 농도로 분사가 가능하며, 에틸렌 가스는 물에 일시적으로 통과한 분사 장치(10)를 통해 스마트-대사체 챔버 내에 고루 분사가 이루어진다. 에틸렌 가스 분사조절은 가스 개폐 장치를 설치하여 조절한다.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.
상기 에틸렌 가스 측정기(11)는 온도조절 팬(7) 아래에 설치되어 챔버부 내부의 에틸렌 가스 농도를 측정한다. 에틸렌 가스 측정기 계량기는 외부 설치하여 측정기로부터 감지되는 양으로 측정한다. 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.
상기 데이터 저장장치 (12)는 챔버부 외부에 설치되어 있으며, 온도 및 습도 자동제어 및 데이터 수집장치(4)와 연결되어 있다. 데이터 수집장치(4)는 컴퓨터일 수 있다. 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.
상기 스마트 대사체 챔버에는 전원공급용 전원 공급장치 (DC24VDC), 전압강하 모듈인 전압분할 장치(VDIV10.1), 데이터 로거와 원격접속을 위한 모뎀 (이더넷 모뎀), 무선 Wifi모뎀 통신을 위한 유무선 공유기인 라우터 및 도조절 팬 / 환기 팬 동작용 전기 제어기가 추가로 설치될 수 있다. 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.
도 2b는 본 발명에 따른 스마트 대사체 챔버의 실제 제작된 제품의 보여주는 사진이다.Figure 2b is a photograph showing the actual manufactured product of the smart metabolite chamber according to the present invention.
참고예 1: 추출물의 제조Reference Example 1: Preparation of extract
이소플라본 측정을 위한 추출물은 수확물을 건조 및 분말화하고, 분말화된 수확물을 각각 10 g 씩 50% 메탄올 400 ml을 넣고 25℃에서 5일간 추출하였다. 상기 추출한 용매를 감압농축하여 16~20%의 수율로 추출물을 얻었다.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%.
참고예 2: 이소플라본 유도체 분석Reference Example 2: Isoflavone derivative analysis
추출물의 이소플라본 유도체 축적정도를 확인하기 위해 HPLC(high performance liquid chromatography) 분석을 수행하였다. High performance liquid chromatography (HPLC) analysis was performed to confirm the degree of accumulation of isoflavone derivatives in the extract.
참고예 1에서와 같이 준비된 추출물을 원심분리기를 이용하여 3500 rpm에서 10분간 원심분리한 후 상등액을 0.45 ㎛ 실린지 필터를 이용하여 여과하였다. 이소플라본 분석을 위해 이용된 기기로는 HPLC는 에이질런트 테크놀로지사(Agilent Technologies Inc., Waldbronn, Germany)의 Agilent 1260 system LC를 이용하였으며, 칼럼은 Lichrophore 100 RP C18 column (4.6×250 mm, 5μm, Merck, Germany)을 검출기는 DAD (diode array detector) 사용하였다. 분석 조건으로는 시료 추출물의 주입량은 20 ㎕을 주입하였고 컬럼온도는 30℃에서 유속은 1 ml/분으로 용매를 흘려보냈으며, 분석 용매는 0.2% 아세트산 수용액(용매 A)를 0.2% 아세트산를 첨가한 아세토니트릴(용매 B) 혼합용액을 사용하여 표 1과 같은 구배 용출(gradient elution)을 적용하여 이소플라본 유도체를 분리하여 254 nm의 검출 파장에서 확인하였다. 정량분석에 이용한 표준용액은 시그마사(Sigma, USA)에서 구입한 순도 98%의 다이드진과 제니스틴, 다이드제인, 제니스테인을 메탄올에 용해시켜 1~100 ㎍/ml 범위로 제조하였다. 제조한 표준용액으로 HPLC 분석을 실시하여 피크 면적(peak area)으로부터 검량선을 작성하였다.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.
시험예 1: 녹두 잎의 에틸렌 처리 최적 조건 분석Test Example 1: Analysis of optimal conditions for ethylene treatment of mung bean leaves
<에틸렌 처리 농도><Ethylene treatment concentration>
녹두 종자 무게에 물을 약 2배 정도 넣은 뒤 20℃에서 15시간 수화시킨 후 암실의 발아기에 넣은 후 20℃에서 24시간 발아시킨 발아된 녹두(발아녹두)를 식물용 포트(pot)에 일반 원예상토를 약 80 g 채운 뒤 2알 내지 3알 파종하였다. 이후 포트를 수직형 식물공장에서 35일간 생육시켰다. 이때 식물공장의 환경으로 광량은 약 13000 LUX 및 약 1200 FC로 설정하고, 온도는 25±0.5℃ 및 습도는 65±5%로 설정하였다. 생육된 녹두를 4그룹으로 나누어 스마트 대사체 챔버로 옮겨 완전히 차단된 밀폐된 공간에서 에틸렌 처리를 진행하였다. 스마트 대사체 챔버의 입구 및 환기 팬 등을 완전히 막은 후 밀폐된 공간으로 만들고 광원조절장치(1)는 백색, 청색 및 적색의 광 비율을 1 : 1 : 1로 설정하고 광량은 400 ㎛ol/m2/s로 설정하고 온도 35±1℃ 및 습도 80±5% 조건에서 생육된 2시간 안정화시킨후, 에틸렌 가스 분사장치(9)를 통하여 에틸렌 가스를 각 그룹에 따라서 0, 2,500, 5,000 및 10,000 ppm의 농도로 24시간 처리 후 수확하였다. 수확된 각 그룹의 녹두 잎을 참고예 1과 같이 추출물을 제조한 후 참고예 2에서와 같은 방식으로 이소플라본 유도체 함량을 분석하여 그 결과를 표 2에 나타냈다. 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.
(㎍/g d.w.)content
(㎍/g dw)
주2)에틸렌 처리는 각 농도로 35도에서 24시간 1회 처리하였다.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.
표 2에 기재된 바와 같이, 에틸렌 농도별(0, 2,500, 5,000 및 10,000 ppm)로 처리된 녹두 잎의 이소플라본 유도체 함량은 에틸렌 농도가 증가할수록 증가하였다. 농도별로 베타-글리코사이드 함량을 기준으로 비교할 시 에틸렌 0 ppm 처리에 비하여 2,500 ppm 처리시는 2.2 배, 5,000 ppm 처리시는 2.6 배, 10,000 ppm 처리시는 2.9배가 증가한 것으로 나타났다.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.
상기와 같은 결과에 의하면 녹두 잎의 에틸렌 처리 농도는 2,500 ~ 10,000 ppm이 적절하며 바람직하게는 5,000 ~ 10,000 ppm이다. 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.
<에틸렌 처리 시간><Ethylene treatment time>
상기와 같이 녹두를 생육하여 4그룹으로 나누어 상기와 같은 환경의 스마트대사체 챔버에서 에틸렌을 10,000 ppm 농도로 각 그룹에 따라 0, 12, 24 및 36시간 처리한 후 수확하였고, 수확된 각 그룹의 녹두 잎을 참고예 1과 같이 추출물을 제조한 후 참고예 2에서와 같은 방식으로 이소플라본 유도체 함량을 분석하여 그 결과를 표 3에 나타냈다. 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.
(㎍/g d.w.)content
(㎍/g dw)
주2)에틸렌 처리는 10,000 ppm 농도로 35도에서 12, 24 및 36시간 1회 처리하였다.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.
표 3에 기재된 바와 같이, 에틸렌 처리 시간이 증가할수록 이소플라본 유도체 함량도 증가하였다. 베타-글이코사이드 총 함량이, 0시간에서 683.28 ㎍/g로 나타났고 이와 비교하여, 12, 24 및 36시간 처리시에는 각각 1,085.67, 4,089.74 및 4,959.27 ㎍/g로 함량이 증가하였고 특히 12시간에서 24시간 사이에 약 3.9배 정도의 함량이 급격히 증가하였다.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.
상기와 같은 결과에 의하면 녹두 잎의 에틸렌 처리 시간은 24 ~ 36시간이 적절하며 가장 바람직하게는 24시간이다. According to the above results, the appropriate ethylene treatment time for mung bean leaves is 24 to 36 hours, most preferably 24 hours.
<에틸렌 처리 횟수><Number of ethylene treatments>
상기와 같이 녹두를 생육하여 4그룹으로 나누어 상기와 같은 환경의 스마트대사체 챔버에서 에틸렌을 10,000 ppm 농도로 각 그룹에 따라 처리 총 시간을 24시간 기준으로 0회, 1회(24시간), 2회(12시간씩) 및 3회(8시간씩) 처리한 후 수확하였고, 수확된 각 그룹의 녹두 잎을 참고예 1과 같이 추출물을 제조한 후 참고예 2에서와 같은 방식으로 이소플라본 유도체 함량을 분석하여 그 결과를 표 4에 나타냈다. 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.
(㎍/g d.w.)content
(㎍/g dw)
주2)에틸렌 처리는 10,000 ppm 농도로 35도에서 24시간 1회 처리, 12시간 2회 처리 및 8시간 3회 처리하였다.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.
표 4에 기재된 바와 같이, 에틸렌 처리 횟수가 증가할수록 이소플라본 유도체 함량도 증가하였다. 총 베타-글리코사이드 함량을 기준으로 비교하였을 시 0, 1, 2 및 3회 처리에서 각각 1,068.25, 5,355.84, 12,902.15 및 13,582.35 ㎍/g로 나타났으며, 특히 2회 및 3회 처리시, 1회 처리시에 비하여 이소플라본 유도체의 함량이 현저히 증가하는 것으로 나타났다.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.
상기와 같은 결과에 의하면 녹두 잎의 에틸렌 처리는 2~3회로 나누어 처리하는 것이 바람직하다. According to the above results, it is desirable to divide the ethylene treatment of mung bean leaves into 2 to 3 treatments.
제조예 1: 수직형 식물공장에서 녹두 재배Manufacturing Example 1: Cultivation of green beans in a vertical plant factory
녹두 종자 (비교예 1) 무게에 물을 약 2배 정도 넣은 뒤 20℃에서 15시간 수화시킨 후 암실의 발아기에 넣은 후 20℃에서 24시간 발아시킨 발아된 녹두(발아녹두)를 준비하였다 (비교예 2).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).
식물용 포트(pot)에 일반 원예상토를 약 80 g 채운 뒤 발아녹두를 2알 내지 3알 파종하였다. 이후 상기에서 파종한 녹두종자의 포트를 수직형 식물공장에서 35일간 생육시켰다 (비교예 3). 이때 식물공장의 환경으로 광량은 약 13000 LUX 및 약 1200 FC로 설정하고, 온도는 25±0.5℃ 및 습도는 65±5%로 설정하였다 (도 1a ~ 도 1c), 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).
상기에 생육된 녹두 식물체를 스마트 대사체 챔버로 옮겨 완전히 차단된 밀폐된 공간에서 에틸렌 처리를 진행하였다. 스마트 대사체 챔버의 입구 및 환기 팬 등을 완전히 막은 후 밀폐된 공간으로 만들고 광원조절장치(1)는 백색, 청색 및 적색의 광 비율을 1 : 1 : 1로 설정하고 광량은 400 ㎛ol/m2/s로 설정하고 온도 35±1℃ 및 습도 80±5% 조건에서 2시간 안정화시킨후, 에틸렌 가스 분사장치(9)를 통하여 에틸렌 가스를 10,000 ppm의 농도로 8시간씩 총 3회 처리한 후 녹두를 수확하였다 (실시예 1; 도 2a ~ 도 2c). 에틸렌 처리와 다음 에틸렌 처리 사이 시간은 2시간 정도로 재배시설을 오픈하여 환기를 시켜 에틸렌을 제거하여 녹두를 안정화시킨다. 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.
비교예 1의 녹두 종자, 비교예 2의 발아 녹두, 비교예 3의 녹두 잎, 실시예 1의 녹두 잎을 각각 흐르는 물에 3회 세척하고 50±2℃에서 24~48시간 건조한 뒤 분말화하였다. 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. .
시험예 2: 이소플라본 HPLC 크로마토그램 분석Test Example 2: Isoflavone HPLC chromatogram analysis
상기 제조예 1에서 제조된 비교예 1, 비교예 2, 비교예 3 및 실시예 1의 분말을 참고예 1과 같이 추출물로 제조한 후, 참고예 2에서와 같이 각각의 이소플라본 유도체 HPLC 크로마토그램 분석 결과를 도 3a ~ 도 3d에 나타냈다. 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.
도 3a ~ 도 3d에 도시된 바와 같이, 비교예 1 및 비교예 2(녹두 종자 및 발아녹두)의 경우는 이소플라본 유도체 피크가 거의 낮거나 불검출되었으며 (도 3a 및 도 3b), 비교예 3(식물공장에서 생육한 녹두 잎)의 에틸렌 무처리 녹두 잎의 경우 6종 이소플라본 유도체(다이드진, 제니스틴, 말로닐 다이드진, 말로닐 제니스틴, 다이드제인 및 제니스테인)의 피크를 확인할 수 있는 정도이며 (도 3c), 실시예 1(본 발명에 따른 에틸렌 처리 녹두 잎)은 6종 이소플라본 유도체의 피크가 현저히 증가됨을 확인할 수 있다 (도 3d). 특히 실시예 1의 녹두 잎은 다이드진(피이크 a), 제니스틴(피이크 b)의 피크 면적이 크게 증가하는 것으로 확인되었다.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).
시험예 3: 이소플라본 유도체의 분리, 정제 및 구조분석Test Example 3: Separation, purification and structural analysis of isoflavone derivatives
실시예 1의 녹두 잎 (도 3d)의 각 피크에 해당하는 이소플라본 유도체에 대해 UPLC-ESI-TOF/MS(Acquity UPLC system coupled with SYNAPT G2-Si HDMS, Waters) 분석을 수행하였다.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).
구체적으로, 참고예 1에서와 같이 준비된 추출물에 대하여 UPLC BEH C18 칼럼(2.1 mm ×100 mm, 1.7 μm, waters)을 이용하고, 용리 조건은 0.1% 포름산 수용액(용매 A)과 0.1% 포름산 아세토니트릴(용매 B) 혼합용액으로 표 5와 같은 구배 용출(gradient elution)을 적용하여 이소플라본 유도체를 분리하였다. 이때 주입된 추출용액의 양은 1 ㎕이며, UPLC-ESI-TOF 질량분석은 표 6과 같은 조건으로 수행하였고, 그 결과를 도 4 ~ 도 9에 나타냈다.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.
도 4에 나타낸 바와 같이, 도 3d의 피크 a의 질량분석 스펙트럼에서 [M+H]-는 415.0 (이론치 분자량: 416.3)로 나타났다. 이로부터 원소 조성이 다이드진의 원소 조성과 일치하는 C21H21O9임을 확인하였다. 피크 a는 참고문헌(J. Agric. Food Chem. 2016, 64, 7315-7324)에 나타난 다이드진과 일치하는바, 화학식 2의 다이드진으로 동정하였다.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.
도 5에 나타낸 바와 같이, 도 3d의 피크 b의 질량분석 스펙트럼에서 [M+H]-는 431.1 (이론치 분자량: 432.3)로 나타났다. 이로부터 원소 조성이 제니스틴의 원소 조성과 일치하는 C21H21O10임을 확인하였다. 피크 b는 참고문헌(J. Agric. Food Chem. 2016, 64, 7315-7324)에 나타난 제니스틴과 일치하는바, 화학식 3의 제니스틴으로 동정하였다.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.
도 6에 나타낸 바와 같이, 도 3d의 피크 c의 질량분석 스펙트럼에서 [M+H]-는 501.4 (이론치 분자량: 502.4)로 나타났다. 이로부터 원소 조성이 말로닐 다이드진의 원소 조성과 일치하는 C24H23O12임을 확인하였다. 피크 c는 참고문헌(J. Agric. Food Chem. 2016, 64, 7315-7324)에 나타난 말로닐 다이드진과 일치하는바, 화학식 4의 말로닐 다이드진으로 동정하였다.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.
도 7에 나타낸 바와 같이, 도 3d의 피크 d의 질량분석 스펙트럼에서 [M+H]-는 517.1 (이론치 분자량: 518.4)로 나타났다. 이로부터 원소 조성이 말로닐 제니스틴의 원소 조성과 일치하는 C24H23O13임을 확인하였다. 피크 d는 참고문헌(J. Agric. Food Chem. 2016, 64, 7315-7324)에 나타난 말로닐 제니스틴과 일치하는바, 화학식 5의 말로닐 제니스틴으로 동정하였다.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.
도 8에 나타낸 바와 같이, 도 3d의 피크 e의 질량분석 스펙트럼에서 [M+H]-는 253.1 (이론치 분자량: 254.2)로 나타났다. 이로부터 원소 조성이 말로닐 다이드진의 원소 조성과 일치하는 C15H10O4임을 확인하였다. 피크 e는 참고문헌(Food Chem. 2020, 305, 125462)에 나타난 말로닐 다이드진과 일치하는바, 화학식 6의 말로닐 다이드진으로 동정하였다.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.
도 9에 나타낸 바와 같이, 도 3d의 피크 f의 질량분석 스펙트럼에서 M+H]-는 269.0 (이론치 분자량: 270.4)로 나타났다. 이로부터 원소 조성이 말로닐 제니스틴의 원소 조성과 일치하는 C15H10O5임을 확인하였다. 피크 f는 참고문헌(Food Chem. 2020, 305, 125462)에 나타난 말로닐 제니스틴과 일치하는바, 화학식 7의 말로닐 제니스틴으로 동정하였다.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.
시험예 4: 이소플라본 유도체의 함량 분석 및 비교Test Example 4: Content analysis and comparison of isoflavone derivatives
참조예 1에서와 같은 방식으로 준비된 실시예 1, 비교예 1, 비교예 2 및 비교예 3의 각 추출물을 0.45㎛의 실린지 필터를 이용하여 여과한 후 참고예 2에서와 같은 방식으로 이소플라본 함량을 분석하고 총량을 정량하여 비교하였고, 그 결과를 표 7 및 도 10에 나타냈다. 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.
(㎍/g d.w.)content
(㎍/g dw)
주2)에틸렌 처리는 10,000 ppm 농도로 35도에서 8시간씩 3회 처리하였다.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.
표 7 및 도 10에 기재된 바와 같이, 비교예 1 및 비교예 2의 녹두 종자 및 발아 녹두는 대부분의 이소플라본 유도체가 검출되지 않았으며, 비교예 3의 에틸렌 무처리 녹두 잎의 경우는 이소플라본 유도체는 검출되지 않거나 소량 검출되었다. 이에 비하여 실시예 1의 녹두 잎의 경우 총 이소플라본 유도체를 14,333.23 ㎍/g 함유하고, 베타-글리코시드 이소플라본 유도체를 13,347.02 ㎍/g 함유하고, 말로일-베타-글로코시드 이소플라본 유도체를 324.98 ㎍/g 함유하고, 아글리콘 이소플라본 유도체를 761.23 ㎍/g 함유하는 것으로 확인되었다. 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.
실시예 1의 이소플라본 유도체 함량은 비교예 3, 비교예 2 및 비교예 1에 비하여 각각 약 14.1배, 약 298.63배 및 약 7,962 배로 현저히 증진되었다. 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.
실시예 1의 녹두 잎에서 다이드진은 11,302.36 ㎍/g, 제니스틴은 1944.66 ㎍/g, 말로닐 다이드진은 110.86 ㎍/g, 말로닐 제니스틴은 214.12 ㎍/g, 다이드제인은 668.99 ㎍/g 및 제니스테인은 92.24 ㎍/g으로 현저히 증진되었다. 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.
시험예 5: 에틸렌 처리 콩잎과의 비교 분석Test Example 5: Comparative analysis with ethylene treated soybean leaves
콩 종자를 이용하여 제조예 1과 동일한 방식으로 콩 식물체를 재배하고 에틸렌 처리하여 콩 잎을 수확한 후 시험예 4와 동일한 방식으로 베타-글리코시드 이소플라본 유도체 함량을 분석하여 시험예 4의 실시예 1의 녹두 잎의 베타-글리코시드 이소플라본 유도체 함량과 비교하였으며 그 결과를 도 11에 나타냈다.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.
도 11에 도시된 바와 같이, 본 발명에 따라 재배된 녹두 잎 (실시예 1)의 다이드진 및 제니스틴 함량은 본 발명에 따라 재배된 콩 잎에 비하여도 각각 7.5배 및 1.45배 높다는 것을 확인할 수 있다. 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.
제조예 2: 약제학적 조성물 제조Preparation Example 2: Preparation of pharmaceutical composition
<1-1> 산제의 제조<1-1> Production of powder
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.1 g, 유당 1.5 g 및 탈크 0.5 gMung 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.
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조하였다.The above ingredients were mixed and filled into an airtight bubble to prepare a powder.
<1-2> 정제의 제조<1-2> Preparation of tablets
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.1 g, 락토오스 7.9 g. 결정성 셀룰로오스 1.5 g 및 마그네슘 스테아레이트 0.5 g0.1 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
상기의 성분들을 혼합한 후 직타법(direct tableting method)으로 정제를 제조하였다.After mixing the above ingredients, tablets were prepared by direct tableting method.
<1-3> 캡슐제의 제조<1-3> Manufacturing of capsules
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.1 g, 옥수수전분 5 g 및 카르복시 셀룰로오스 4.9 gMung 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
상기의 성분들을 혼합하여 분말을 제조한 후, 상기 분말을 통상의 캡슐제의 제조방법에 따라 경질 캡슐에 충전하여 캡슐제를 제조하였다.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> 주사제의 제조<1-4> Preparation of injections
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.1 g, 주사용 멸균 증류수 적량, pH 조절제 적량Mung 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
통상의 주사제의 제조방법에 따라 1 앰플 당(2 ㎖) 상기의 성분 함량으로 제조하였다.It was prepared with the above ingredients per 1 ampoule (2 ml) according to the usual manufacturing method for injections.
제조예 3: 건강기능성 식품의 제조Manufacturing Example 3: Manufacturing of health functional food
<1-1> 혼합차의 제조<1-1> Manufacturing of mixed tea
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물과 향미를 갖는 허브 식물체를 혼합하여, 혼합차를 제조하였다.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> 밀가루 식품의 제조<1-2> Manufacturing of flour food
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.5 ~ 5.0 중량부를 밀가루에 첨가하고, 이를 혼합한 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하였다.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> 스프 및 육즙(gravies)의 제조<1-3> Preparation of soups and gravies
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 0.1 ~ 5.0 중량부를 스프 및 육즙에 첨가하여 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조하였다.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> 그라운드 비프(ground beef)의 제조<1-4> Production of ground beef
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 10 중량부를 그라운드 비프에 첨가하여 건강 증진용 그라운드 비프를 제조하였다.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> 유제품(dairy products)의 제조<1-5> Manufacturing of dairy products
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 5 ~ 10 중량부를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.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> 선식의 제조<1-6> Manufacturing of Seonsik
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화한 후 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.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.
검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.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.
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물을 진공 농축기에서 감압농축하고, 분무, 열풍건조기로 건조하여 얻은 건조물을 분쇄기로 입도 60 메쉬로 분쇄하여 건조분말을 얻었다.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.
상기에서 제조한 곡물류, 종실류 및, 본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물을 하기의 비율로 배합하여 제조하였다.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.
곡물류(현미 30 중량부, 율무 15 중량부, 보리 20 중량부), 종실류(들깨 7 중량부, 검정콩 8 중량부, 검정깨 7 중량부), 본 발명의 이소플라본 유도체가 강화된 콩잎, 콩 줄기 및 콩 뿌리, 이의 추출물 또는 추출물의 분획물(3 중량부), 영지(0.5 중량부), 지황(0.5 중량부)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> 음료의 제조<1-7> Manufacturing of beverages
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 100 mg, 구연산 100 mg, 올리고당 100 mg, 매실농축액 2 mg, 타우린 100 mg, 정제수를 가하여 전체 500 mlMung 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.
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 1 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관하여 본 발명의 건강음료 조성물 제조에 사용한다.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.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였으나, 수요계층, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합 비를 임의로 변형 실시하여도 무방하다.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.
제조예 4: 화장품 제조Manufacturing Example 4: Cosmetic manufacturing
<1-1> 크림 제조<1-1> Cream production
본 발명의 이소플라본 유도체가 증진된 녹두 잎, 이의 추출물 또는 추출물의 분획물 4.6 중량부, 세토스테아릴알코올 2.8 중량부, 밀납 2.6 중량부, 스테아린산 1.4 중량부, 친유형모노스테아린산글리세린 2 중량부, 피이지-100 스테아레이트 1 중량부, 세스퀴올레인산소르비탈 1.4 중량부, 호호바오일 4 중량부, 스쿠알란 3.8 중량부, 폴리소르베이트 60 1.1 중량부, 마카다미아오일 2 중량부, 초산토코페롤 0.2 중량부, 메칠폴리실록산 0.4 중량부, 에칠파라벤 0.1 중량부, 프로필파라벤 0.1 중량부, Euxyl K-400 0.1 중량부, 1,3-부틸렌글리콜 7 중량부, 메칠파라벤 0.05 중량부, 글리세린 6 중량부, d-판테놀 0.2 중량부, 트리에탄올아민 0.2 중량부, pt 41891 0.2 중량부, p-H2O 46.05 0.295 중량부를 혼합하여 크림 제조방법에 따라 제조하였다.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> 로션의 제조<1-2> Production of lotion
본 발명의 이소플라본 유도체가 증진된 녹두 잎의 추출물 또는 추출물의 분획물 3.5 중량부, 세토스테아릴알코올 1.6 중량부, 스테아린산 1.4 중량부, 친유형모노스테아린산글리세린 1.8 중량부, 피이지-100 스테아레이트 2.6 중량부, 세스퀴올레인산소르비탈 0.6 중량부, 스쿠알렌 4.8 중량부, 마카다미아오일 2 중량부, 호호바오일 2 중량부, 초산토코페롤 0.4 중량부, 메칠폴리실록산 0.2 중량부, 에칠파라벤 0.1 중량부, 프로필파라벤 0.1 중량부, 1,3-부틸렌글리콜 4 중량부, 메칠파라벤 0.1 중량부, 산탄검 0.1 중량부, 글리세린 4 중량부, d-판테놀 0.15 중량부, 알란토인 0.1 중량부, 카르보내(2% aq. Sol) 4 중량부, 트리에탄올아민 0.15 중량부, 에탄올 3 중량부, pt 41891 0.1 중량부, p-H20 48.3 중량부를 혼합하여 로션 제조방법에 따라 제조하였다.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: 광원조절장치 (적색, 청색, 백색 광원)
2: 광원조절장치 (UV-A 혹은 UV-B)
3: LED 광원부 (적색/청색/백색/UV-A 혹은 B)
4: 온도 및 습도 자동제어 및 데이터 수집 장치
5: 온도, 습도 및 이산화탄소 측정기
6: 환기 팬
7: 온도조절 팬
8: 온도조절용 물 순환장치
9: 에틸렌 가스통
10: 에틸렌 가스 분사장치
11: 에틸렌 가스 측정기
12: 데이터 저장장치
13: 식물생육 관찰 창1: 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)
[화학식 1]
상기 식에서, R1은 H 또는 OH이고, R2는 글루코오스 또는 글루코오스 말로네이트이고,
상기 방법은
i) 재배 시설에서 녹두 종자 또는 발아된 녹두를 파종하여 20 ~ 40일을 생육시키는 단계;
ii) 생육된 녹두에 2,500 ~ 10,000 ppm의 농도의 에틸렌을 8 ~ 12 시간씩 2 ~ 3회 처리하는 단계; 및
iii) 녹두 잎을 수확하는 단계;를 포함하는 것인 방법.
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.
상기 약제학적 조성물은 에스트로겐 결핍에 기인한 질환의 예방 및 치료용인 것인 약제학적 조성물.
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.
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