KR101823525B1 - Antibacterial composition for inhibiting Pectobacterium carotovorum subsp. Carotovorum comprising Suaeda japonica extracts as an active ingredient - Google Patents
Antibacterial composition for inhibiting Pectobacterium carotovorum subsp. Carotovorum comprising Suaeda japonica extracts as an active ingredient Download PDFInfo
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- KR101823525B1 KR101823525B1 KR1020170174187A KR20170174187A KR101823525B1 KR 101823525 B1 KR101823525 B1 KR 101823525B1 KR 1020170174187 A KR1020170174187 A KR 1020170174187A KR 20170174187 A KR20170174187 A KR 20170174187A KR 101823525 B1 KR101823525 B1 KR 101823525B1
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Abstract
Description
본 발명은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는, 배추 무름병 원인균 억제용 항균 조성물에 관한 것이다. 보다 상세하게는, 칠면초 에탄올 추출물로부터 수득한 헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, 및 부탄올 분획물의 배추 무름병 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC) 억제용 항균 조성물, 상기 조성물을 포함하는 농업용 작물 보호제, 상기 조성물을 포함하는 식품 세척제, 상기 조성물을 포함하는 식품 첨가제 및 상기 조성물을 이용한 배추 무름병 원인균 방제 방법에 관한 것이다. 또한, 상기 칠면초 추출물을 유효성분으로 포함하는 펙토박테리움 카로토보룸 증식 억제 또는 사멸용 조성물에 관한 것이다.The present invention relates to an antimicrobial composition for inhibiting root rot caused by a Chinese cabbage ( Suaeda japonica ) extract as an active ingredient. More particularly, the present invention relates to an antimicrobial composition for inhibiting Pectobacterium carotovorum subsp. Carotovorum (PCC), which is a causative agent of cabbage root rot, of hexane fraction, chloroform fraction, ethyl acetate fraction and butanol fraction obtained from an ethanol extract of chile, A food cleaner comprising the composition, a food additive containing the composition, and a method for controlling causative agents of cabbage acne causing bacteria using the composition. Further, the present invention relates to a composition for inhibiting or preventing the growth of Pectobacterium carotovorum, which comprises the above-mentioned extract of chile peony root as an active ingredient.
세균성 무름병은 주요 채소작물에 병을 일으킨다. 무름병은 독특한 냄새가 나면서 흐물흐물해져서 썩는 식물의 병해를 지칭하는 것으로, 흔히 연부병(軟腐病)이라고도 한다. 무름병에 걸리면 세포벽 중간층의 펙틴질이 녹아 처음에는 물기가 보이는 것에 그치나, 점차로 물러져 썩고 액체처럼 흐물흐물해진다. 무름병은 채소작물의 이동 보관 시에 큰 손실을 일으키며, 배추에 많이 나타나는 병해이나, 그 밖에 토마토, 고구마, 감자에도 많이 나타난다.Bacterial rotting disease causes disease in major vegetable crops. Grape rot disease is a disease that rotates due to unusual smell and stains, often called soft rot. When it gets cloudy, the pectin in the middle layer of the cell wall melts, and at first it looks like water, but it gradually recovers and rotates and becomes like liquid. Grape rot disease causes a great loss in storage and transportation of vegetable crops, and many diseases appear in cabbage, but also in tomatoes, sweet potatoes and potatoes.
배추 무름병은 세균에 의한 병해로 고온다습 조건에서 발병이 잘 되고 발병하면 방제가 매우 어려운 식물병 중 하나이다. 배추 무름병은 최근 기후변화에 의해 고온다습 환경조건이 형성되면서 배추 농가에 많은 피해를 주고 있다(농촌진흥청, 1997). 배추 무름병에 관여하는 병원균은 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)으로 보고(한국식물병리학회, 2009)되어 있으며, 이러한 병원균은 펙테이트리에이즈(pectate lyase)라는 효소를 생산하여 식물체의 세포벽을 붕괴시키는 특성을 가지고 있다. 펙토박테리움 카로토보룸 아종 카로토보룸은 생장 적온이 26~28℃이나, 다른 식물 병원 세균과는 달리 상온인 36℃ 정도에서도 잘 자라, 여름철 배추 생산에 가장 큰 장애 요인이다. 무름병균은 배추를 비롯하여 거의 모든 식물에 침입하여 발병할 수 있는데, 한번 침입하면 주변의 다른 채소로 옮겨가 발병을 전파한다. 또한, 채소의 재배 시기뿐 아니라 이동 보관 시에도 신선 채소 등에 무름병이 발병한다. 이러한 채소 무름병은 신선 채소 재배 및 유통에서 가장 피해가 큰 병원균으로 오랫동안 알려져 왔으나 병원균인 펙토박테리움 카로토보룸 아종 카로토보룸은 토양에서 비교적 오랫동안 생존이 가능하며, 농약에 의한 방제 효과도 일반적으로 낮아 새로운 방제법 개발이 요구되고 있다.Cabbage rot disease is a disease caused by bacteria that is well tolerated under high temperature and high humidity conditions and is one of the plant diseases that are very difficult to control when it develops. In recent years, cabbage blight has been causing many damage to cabbage farms due to the formation of high temperature and high humidity environment conditions due to climate change (RDA, 1997). The pathogenic bacteria responsible for cabbage blight are Pectobacterium carotovorum subsp. Carotovorum (Korean Society of Plant Pathology, 2009), and these pathogens are called pectate lyase It has the property of disrupting the cell walls of plants by producing enzymes. Pectobacterium carotovorum The subspecies of carotovorum are 26-28 ℃ in growth temperature, but unlike other botanical germs, it grows well at 36 ℃, which is the biggest obstacle to the production of cabbage in summer. Dried bacillus can invade almost all plants including cabbage, and once it breaks in, it moves to other vegetables in the vicinity and spreads the disease. In addition, not only when the vegetables are grown, but also when they are stored for moving, there is a risk of redness in fresh vegetables. These vegetable rot ones have long been known as the most harmful pathogens in fresh vegetable cultivation and distribution. However, the pectobacterium carotovorum, which is a pathogenic bacterium, is able to survive for a relatively long time in the soil, It is required to develop a new control method.
배추 무름병 방제 약제로 등록되어 있는 화학 농약은 가스가마이신 액제 등 16종이 등록(한국작물보호협회, 2012)되어 있으나, 대부분이 농용항생제(agricultural antibiotics)가 주성분으로 되어있어 우리가 식품으로 먹는 배추에 장기적인 사용으로 인한 항생제 내성 문제 등이 대두되고 있으며, 무름병에 대한 방제 효과 또한 낮은 점이 농가에서 많은 어려움을 느끼고 있는 부분이다.We have registered 16 kinds of chemical pesticides such as gas-liquid liquor (Korea Crop Protection Association, 2012), but most of them are agricultural antibiotics. Antibiotics resistance due to long-term use, and the low control effect against erosion is a difficult part of farming.
식물병 방제를 위해 사용하는 항생제 또는 화학 물질은 농업 생태계에서 항생제 저항성의 유발 및 환경에 대한 오염 및 잔류 독성의 문제를 일으킨다. 이에, 환경 친화적이면서도 인축에 대한 독성이 거의 없는 배추 무름병의 방제제 개발의 필요성이 절실히 요구되고 있다.Antibiotics or chemicals used to control plant diseases cause problems with antibiotic resistance in the agricultural ecosystem, environmental pollution and residual toxicity. Therefore, it is urgently required to develop a control agent for Chinese cabbage blight which is environmentally friendly and has little toxicity to human skin.
최근 배추 무름병에 대한 친환경 방제법 개발이 시급히 요구되어 연구 개발이 점차 이루어지고 있는데, 국내에서의 무름병 친환경 방제법 연구는 미네랄 오일에 의한 방제(Lee, S.H. et al., Phytopathology, 91:S53-S54, 2001) 등이 보고되어 있고, 배추 무름병 생물적 방제 연구로는 3종의 유용세균 Lactobacillus KLF01, Lactococcus KLC02, 및 Paenibacillus KPB3 균주를 이용한 생물적 방제연구(Anupama Shrestha et al., The Korean Journal of Pesticide Science, 13(4):325-331, 2009) 등이 있을 뿐 아직 많은 연구 결과가 보고되지 않고 있다.Recently, the development of environment-friendly control method for cabbage rot disease has been urgently required and research and development has been progressing gradually. The research on environment control of rottling disease in Korea has been conducted by controlling mineral oil (Lee, SH et al ., Phytopathology , 91: S53-S54, 2001 ), And bioassay studies using three species of useful bacteria, Lactobacillus KLF01, Lactococcus KLC02, and Paenibacillus KPB3 (Anupama Shrestha et al. , The Korean Journal of Pesticide Science , 13 (4): 325-331, 2009), but many studies have not yet been reported.
한편, 염생식물은 염류가 함유된 토지에서 생육할 수 있는 식물로, 바닷물과 담수가 반복적으로 교차하는 지역에서 생육하게 되면서, 육상 식물과 해상 식물이 가진 다양한 생리 활성 물질들을 동시에 포함하는 것으로 추정되고 있다. 국내의 경우 퉁퉁마디(함초)(Saliconia herbacea), 나문재(Suaeda asparagoides), 칠면초(Suaeda japonica), 갯질갱(Limonium tetragonum), 해당화(Rosa rugosa), 통보리사초(Carex kobomugi), 수송나물(Salsolakormarovi Iljin), 해홍나물(Suaeda maritima) 등이 대표적인 염생식물로 알려져 있다. 이러한 염생식물은 최근 식품 가공 기술의 발달로 인해, 다양한 먹거리로 이용되며, 또한 식물성 소금 제조원으로 각광받고 있다. 현재까지는 염생식물을 이용하여 주로 식품 원재료 이용 및 소금 제조가 가장 일반적으로 이루어지고 있으며, 대표적인 염생식물인 함초, 나문재, 해홍나물, 칠면초 등의 항산화, 항균, 항염증 등 다양한 생리 활성 효과에 관해 보고되고 있다.On the other hand, it is presumed that the halophyte is a plant that can grow on the soil containing salt, and it grows in a region where sea water and fresh water repeatedly intersect, and simultaneously contains various physiologically active substances of land plants and marine plants have. In Korea, Salmonia herbacea), namunjae (Suaeda Asparagoides , Suaeda japonica , Limonium tetragonum , Rosa rugosa , Carex kobomugi , Salsolakormarovi Iljin , and Suaeda maritima are known as typical halophytes. These halophilic plants have recently been used as a variety of foods due to the development of food processing technology, and are also attracting attention as a source of vegetable salt. So far, the most common use of food ingredients and salt production is by using halophytes, and various physiological activities such as antioxidation, antibacterial and anti-inflammatory effects of representative halophilic plants such as green tea, .
칠면초(Suaeda japonica)는 명아주과(Chenopodiaceae)에 속하는 일년생 초본식물로 고농도의 염류 토양에서 자라는 호염성 식물이며, 바닷물 속에 포함되어 있는 각종 미네랄이 다량 함유되어 있는 것으로 보고되었다. 특히, 칠면초는 함초보다 Na+ 함량은 적은 반면 Ca2 +, Fe2 +, Mn2 +은 함초보다 더 많은 양이 함유되어 있다고 보고되고 있다. 한편, Ca은 골다공증의 예방과 치료, 신경전달활성, 암예방 및 치료, 고혈압 치료, 콜레스테롤 수치 저하 및 성인병 예방과 관절염 예방 치료, 피부 노화 방지 등의 다양한 기능을 갖고 있는 것으로 보고되고 있다. Suaeda japonica is an annual herbaceous plant belonging to Chenopodiaceae. It is a horticultural plant growing in a high concentration of salt soil and reported to contain a large amount of various minerals contained in seawater. In particular, S. japonica is Na + content is less than Salicornia while Ca 2 +, Fe 2 +, Mn 2 + has been reported to contain a greater amount than the Salicornia. On the other hand, Ca has been reported to have various functions such as prevention and treatment of osteoporosis, neurotransmitter activity, prevention and treatment of cancer, treatment of hypertension, reduction of cholesterol level, prevention of adult diseases, prevention of arthritis and prevention of skin aging.
그러나, 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 칠면초(Suaeda japonica)의 항균 효과에 대해서는 현재까지 전혀 보고된 바 없다.However, the antimicrobial effect of Suaeda japonica against Pectobacterium carotovorum subsp. Carotovorum (PCC), a cause of cabbage root rot , has not been reported to date.
이러한 배경하에서, 본 발명자들은 배추 무름병균에 대한 항균 활성을 나타내면서, 향후 절임배추, 김치 등 식품에 적용하였을 시, 인체에 안전하고 친환경적인 항균 조성물을 개발하고자 예의 연구노력한 결과, 약용식물로서 사용되어 온 염생식물인 칠면초(Suaeda japonica)의 추출물, 특히 칠면초 에탄올 추출물로부터 수득한 헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, 및 부탄올 분획물이 배추 무름병균에 대한 생장 억제 및 살균 효과가 우수함을 확인함으로써, 본 발명을 완성하게 되었다.Under these circumstances, the inventors of the present invention have made efforts to develop a safe and environmentally friendly antimicrobial composition for human body when they are applied to foods such as pickled cabbage, kimchi and the like, showing antimicrobial activity against Chinese cabbage root rot, It was confirmed that the extracts of Suaeda japonica , especially hexane fraction, chloroform fraction, ethyl acetate fraction, and butanol fraction obtained from the chrysanthemum ethanol extract, were excellent in growth inhibition and bactericidal effect against the cabbage root rot, Thereby completing the invention.
본 발명의 목적은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는, 배추 무름병 원인균 억제용 항균 조성물을 제공하기 위한 것이다.It is an object of the present invention to provide an antimicrobial composition for inhibiting root rot caused by a Chinese cabbage ( Suaeda japonica ) extract as an active ingredient.
본 발명의 다른 목적은 상기 조성물을 포함하는 농업용 작물 보호제, 식품 세척제 및 식품 첨가제를 제공하기 위한 것이다.Another object of the present invention is to provide an agricultural crop protection agent, a food cleaner and a food additive comprising the composition.
본 발명의 또 다른 목적은 상기 조성물을 이용하여 배추 무름병 원인균을 방제하는 방법을 제공하기 위한 것이다.It is still another object of the present invention to provide a method for controlling cabbage root rot caused by using the composition.
본 발명의 또 다른 목적은 상기 칠면초 추출물을 유효성분으로 포함하는, 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum) 증식 억제 또는 사멸용 조성물을 제공하기 위한 것이다.It is still another object of the present invention to provide a method for producing Pectobacterium tuberculosis, carotovorum subsp. Carotovorum ) proliferation inhibiting or killing composition.
상기 목적을 달성하기 위하여, 본 발명은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는, 배추 무름병 원인균 억제용 항균 조성물, 상기 조성물을 포함하는 농업용 작물 보호제, 상기 조성물을 포함하는 식품 세척제, 상기 조성물을 포함하는 식품 첨가제 및 상기 조성물을 이용한 배추 무름병 원인균 방제 방법을 제공한다. 또한, 상기 칠면초 추출물을 유효성분으로 포함하는 펙토박테리움 카로토보룸 증식 억제 또는 사멸용 조성물을 제공한다.In order to achieve the above object, the present invention provides an antimicrobial composition for inhibiting root rot caused by a Chinese cabbage comprising Suaeda japonica extract as an active ingredient, an agricultural crop protection agent containing the composition, a food cleaner containing the composition, And a method for controlling a causative agent of cabbage acne causing bacteria using the composition. In addition, the present invention provides a composition for inhibiting or inhibiting proliferation of Pectobacterium carotovorum, which comprises the above extract of chile peony root as an active ingredient.
이하, 본 발명을 구체적으로 설명한다.Hereinafter, the present invention will be described in detail.
하나의 양태로서, 본 발명은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는, 배추 무름병 원인균 억제용 항균 조성물을 제공한다.In one embodiment, the present invention provides an antimicrobial composition for inhibiting root rot caused by a Chinese cabbage comprising Suaeda japonica extract as an active ingredient.
본 발명의 용어 "칠면초(Suaeda japonica)"는 명아주과의 일년생 식물로, 우리나라의 서남해안 조간대에 자생하는 염생식물(halophyte)로, 성장시기에 따라 녹색에서 적색까지 다양한 색을 띈다. 한편, 칠면초는 적색을 띄고 있어 식물성 소금인 홍염과 천연 염료로써 이용되고 있으며, 한방에서는 지상부를 해열제로 사용하고 있다. 또한, 칠면초는 고농도의 NaCl 함량과 마그네슘, 칼슘, 칼륨 등의 미네랄이 풍부한 것으로 알려져 있다. 칠면초의 생리 활성에 대한 연구로는 항산화 효과가 있다고 보고되어 있을 뿐, 다양한 생리활성 및 유용성분에 대한 연구는 매우 미흡한 실정이다. 아울러, 칠면초는 수확량이 적은 함초를 비롯한 다른 염생식물들과 달리 갯벌 및 염습지 등의 조간대에 넓게 분포되어 있어 생물자원으로 활용할 경우 대량 원료 확보가 가능하다.The term " Suaeda japonica "of the present invention refers to a perennial plant of the Japanese apricot. It is a halophyte native to the intertidal zone of the west coast of Korea. It has various colors ranging from green to red depending on the growing season. On the other hand, the chili pepper is red, and it is used as vegetable salt red flour and natural dyestuff. Also, it is known that the chili pepper is rich in high NaCl content and minerals such as magnesium, calcium, and potassium. Studies on the physiological activities of the pyrethroids have been reported to have antioxidative effects, but studies on various physiological activities and useful constituents are insufficient. In addition, unlike other halophytes, which are less harvested, chilgumcho is widely distributed in tidal flats and salt marshes, and thus it is possible to secure bulk materials when used as biological resources.
본 발명의 용어 "배추 무름병(bacterial soft rot)"은 세균에 의한 병해로 고온다습 조건에서 발병이 잘 되고 발병하면 방제가 매우 어려운 식물병 중 하나이다. 배추 무름병에 관여하는 병원균은 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)으로 보고(한국식물병리학회, 2009)되어 있으며, 이러한 병원균은 펙테이트리에이즈(pectate lyase)라는 효소를 생산하여 식물체의 세포벽을 붕괴시키는 특성을 가지고 있다. 펙토박테리움 카로토보룸 아종 카로토보룸은 생장 적온이 26~28℃이나, 다른 식물 병원 세균과는 달리 상온인 36℃ 정도에서도 잘 자라, 여름철 배추 생산에 가장 큰 장애 요인이다. 무름병균은 배추를 비롯하여 거의 모든 식물에 침입하여 발병할 수 있는데, 한번 침입하면 주변의 다른 채소로 옮겨가 발병을 전파한다.The term " bacterial soft rot "of the present invention is one of botanical bottles that is well tolerated under high temperature and high humidity conditions due to bacterial disease and is very difficult to control when it develops. The pathogenic bacteria responsible for cabbage blight are Pectobacterium carotovorum subsp. Carotovorum (Korean Society of Plant Pathology, 2009), and these pathogens are called pectate lyase It has the property of disrupting the cell walls of plants by producing enzymes. Pectobacterium carotovorum The subspecies of carotovorum are 26-28 ℃ in growth temperature, but unlike other botanical germs, it grows well at 36 ℃, which is the biggest obstacle to the production of cabbage in summer. Dried bacillus can invade almost all plants including cabbage, and once it breaks in, it moves to other vegetables in the vicinity and spreads the disease.
상기 칠면초(Suaeda japonica) 추출물은 하기의 단계들을 포함하는 제조방법에 의해 제조되는 것이 바람직하나 이에 한정되지 않는다:The Suaeda japonica extract is preferably, but not exclusively, prepared by a method comprising the steps of:
1) 칠면초(Suaeda japonica)에 추출용매를 가하여 추출하는 단계;1) extracting Suceeda japonica with an extraction solvent;
2) 단계 1)의 추출물을 여과하는 단계;2) filtering the extract of step 1);
3) 단계 2)의 여과한 추출물을 감압 농축한 후 건조하여 칠면초의 추출물을 제조하는 단계; 및3) concentrating the filtered extract of step 2) under reduced pressure and drying to prepare an extract of chile; And
4) 단계 3)의 칠면초 추출물을 추가적으로 유기용매로 추출하여 칠면초 분획물을 제조하는 단계.4) Extracting the extract of Cheil cunea in step 3) with an organic solvent to prepare Cheil side fraction fraction.
상기 방법에 있어서, 단계 1)의 칠면초(Suaeda japonica)는 재배한 것 또는 시판되는 것 등 제한 없이 사용할 수 있다. 또한, 단계 1)에서 이에 제한되지는 않으나, 칠면초는 줄기, 가지, 잎 또는 뿌리를 모두 포함하여 추출하는 것일 수 있다.In the above method, the step ( Suaeda japonica ) of step 1) can be used without limitation such as cultivated or commercially available. In addition, although not limited in step 1), the pyramid may be extracted including stem, branch, leaf or root.
상기 방법에 있어서, 상기 단계 1)의 추출용매는 물, 알코올 또는 이들의 혼합물을 사용하는 것이 바람직하다. 상기 알코올로는 C1 내지 C4 저급 알코올을 이용하는 것이 바람직하며, 저급 알코올로는 에탄올을 이용하는 것이 바람직하다. 추출방법으로는 진탕추출, 속슬렛(Soxhlet) 추출 또는 환류 추출을 이용하는 것이 바람직하나 이에 한정되지 않는다. 상기 추출용매를 건조된 칠면초 시료 분량에 1 내지 10배 첨가하여 추출하는 것이 바람직하다. 추출온도는 20℃ 내지 100℃ 인 것이 바람직하고, 20℃ 내지 40℃인 것이 더욱 바람직하고, 실온인 것이 가장 바람직하나, 이에 한정하지 않는다. 또한, 추출시간은 8 내지 24시간인 것이 바람직하며, 10 내지 15시간인 것이 더욱 바람직하고, 12시간인 것이 가장 바람직하나, 이에 한정하지 않는다. 아울러, 추출 횟수는 1 내지 5회인 것이 바람직하며, 3 내지 4회 반복 추출하는 것이 더욱 바람직하고, 3회인 것이 가장 바람직하나, 이에 한정되는 것은 아니다. In the above method, it is preferable to use water, an alcohol or a mixture thereof in the extraction solvent of the step 1). As the alcohol, a C 1 to C 4 lower alcohol is preferably used, and as the lower alcohol, ethanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. It is preferable that the extraction solvent is added by 1 to 10 times the amount of the dried pyramid sample. The extraction temperature is preferably 20 占 폚 to 100 占 폚, more preferably 20 占 폚 to 40 占 폚, and most preferably room temperature, but is not limited thereto. The extraction time is preferably 8 to 24 hours, more preferably 10 to 15 hours, most preferably 12 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but is not limited thereto.
상기 방법에 있어서, 단계 3)의 감압농축은 진공감압농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나 이에 한정하지 않는다.In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.
상기 방법에 있어서, 단계 4)의 유기용매는 노르말-헥산(n-Hexane), 클로로포름(chloroform), 에틸아세테이트(EtOAc) 또는 부탄올(BuOH)인 것이 바람직하나, 이에 한정하지 않는다. 상기 분획물은 칠면초 추출물을 물에 현탁시킨 후 노르말-헥산(n-Hexane), 클로로포름(chloroform), 에틸아세테이트(EtOAc), 노르말-부탄올(n-BuOH) 및 물(H2O)로 순차적으로 계통 분획하여 수득한 노르말-헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, 노르말-부탄올 분획물 또는 물 분획물 중 어느 하나인 것이 바람직하나, 이에 한정하지 않는다. 상기 분획물은 상기 칠면초 추출물로부터 분획 과정을 1 내지 5회, 바람직하게는 3회 반복하여 수득할 수 있고, 분획 후 감압 농축하는 것이 바람직하나 이에 한정하지 않는다.In the method, the organic solvent of step 4) is n-hexane (n -Hexane), chloroform (chloroform), one preferably in ethyl acetate (EtOAc) or butanol (BuOH), but not always limited thereto. The fraction was suspended in water, the S. japonica extract n-hexane (n -Hexane), chloroform (chloroform), ethyl acetate (EtOAc), n-Butanol (n -BuOH) and water (H 2 O) to the grid sequentially But is not limited to, the n-hexane fraction, the chloroform fraction, the ethyl acetate fraction, the n-butanol fraction or the water fraction obtained by fractionation. The fractions can be obtained by repeating the
본 발명의 용어 "항균"은 세균이나 진균 등의 미생물의 생장 및 증식을 억제 또는 제어하는 작용을 의미하는 것으로, 본 발명의 미생물은 이에 제한되지는 않으나, 배추 무름병 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)일 수 있다.The term "antibacterial" of the present invention means an action of inhibiting or controlling the growth and proliferation of microorganisms such as bacteria and fungi. The microorganisms of the present invention include, but are not limited to, a bacterial strain, Pectobacterium carotovorum subsp. Carotovorum .
상기 배추 무름병 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)을 방제하기 위해 가스가마이신 액제와 같은 화학농약이 사용되고 있으나, 대부분이 농용항생제(agricultural antibiotics)가 주성분으로 되어있어 식품으로 먹는 배추에 장기적으로 사용할 경우 항생제 내성 문제 등이 발생할 수 있으므로 비교적 인체에 무해한 물질을 사용하는 것이 중요하다.In order to control Pectobacterium carotovorum subsp. Carotovorum , which is a causative agent of the cabbage root rot, chemical pesticides such as gas lecithin solution are used, but most of them are agricultural antibiotics. It is important to use relatively harmless substances because it may cause problems such as antibiotic resistance when used for long term in Chinese cabbage to eat.
따라서, 본 발명의 칠면초(Suaeda japonica) 추출물, 특히 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)을 농업용 작물 보호제, 식품 세척제 또는 식품 첨가제 등에 포함시키면 배추 무름병을 유발하는 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)의 생장 억제 및 사멸 효과를 달성함으로써, 배추 무름병 발생을 예방 및 억제/제어할 수 있다.Therefore, the invention of S. japonica (Suaeda japonica) extract, particularly obtained from an ethanol extract of S. japonica a n-hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction ( Pectobacterium carotovorum subsp. Carotovorum ), which is a causative organism of cabbage blight, can be obtained by incorporating the n- butanol fraction into agricultural crop protection agent, food cleanser or food additive, It is possible to prevent and inhibit / control the occurrence of Chinese cabbage blight.
본 발명의 일실시예에서, 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)의 항균 효과를 확인하기 위하여, 페이퍼 디스크 확산법(paper disc diffusion method)을 이용하여 배추 무름병 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 항균 활성을 측정한 결과, 생육저지환(clear zone)의 직경 크기가 상기 4가지 모든 분획물에서 PCC 균주에 대하여 생장 저해 활성이 우수함을 확인하였다(표 1).In one embodiment of the invention, a S. japonica n obtained from the ethanol extract of hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction (n -butanol fraction ( Pectobacterium carotovorum subsp. Carotovorum (PCC)), a causative organism of cabbage root rot, was examined using paper disc diffusion method. As a result, the antimicrobial activity of Pectobacterium carotovorum subsp. It was confirmed that the diameter of the clear zone of the growth was superior to that of the PCC strain in all four fractions (Table 1).
다른 하나의 양태로서, 본 발명은 상기 배추 무름병 원인균 억제용 항균 조성물을 포함하는 농업용 작물 보호제를 제공한다.In another aspect, the present invention provides an agricultural crop protection agent comprising the antimicrobial composition for inhibiting the causative agent of cabbage acne-causing bacteria.
본 발명에 있어서, 상기 농업용 작물 보호제는 칠면초(Suaeda japonica) 추출물, 특히 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)을 유효성분으로 포함할 수 있다. 상기 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, n-부탄올 분획물을 유효성분으로 첨가하여 제조한 농업용 작물 보호제는 배추 무름병을 유발하는 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)의 생장을 억제하거나 사멸시켜, 배추 무름병 발생을 예방 및 억제/제어할 수 있다.In the present invention, wherein the agricultural crop protection is S. japonica (Suaeda japonica) extract, particularly obtained from an ethanol extract of S. japonica a n-hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction) , n - butanol fraction (n -butanol fraction) may comprise as the active ingredient. The agricultural crop protection agent prepared by adding the n - hexane fraction, the chloroform fraction, the ethyl acetate fraction and the n - butanol fraction obtained as an active ingredient to the agricultural chemical crop protection agent obtained from the chllycolipid ethanol extract, Inhibit or control the growth of Pectobacterium carotovorum subsp. Carotovorum ), preventing, inhibiting / controlling the occurrence of cabbage blight disease .
본 발명에 있어서, 상기 농업용 작물 보호제는 부형제를 추가로 포함할 수도 있다. 상기 부형제는 통상의 미생물 제제, 항균 효과 증진제, 희석제 또는 담체일 수 있다. 한편, 상기 농업용 작물 보호제는 상기 유효성분 이외에도 약효를 증진시키거나 적용범위를 확대시킬 목적으로 기존에 상업화되어 사용되고 있거나 개발되어 있는 다른 살균제, 살충제, 제초제, 식물생장조절제 또는 비료를 일정 함량 범위로 추가로 포함시켜 혼합제 형태로 제제화할 수 있다.In the present invention, the agricultural crop protection agent may further comprise an excipient. The excipient may be a conventional microbial agent, an antimicrobial effect enhancer, a diluent or a carrier. Meanwhile, the agricultural crop protection agent may be added to other agricultural chemicals, pesticides, herbicides, plant growth regulators or fertilizers in a predetermined amount range for the purpose of improving the efficacy or expanding the application range in addition to the above- To form a mixture in the form of a mixture.
본 발명에 있어서, 상기 농업용 작물 보호제에 포함되는 부형제, 희석제는 농업 분야에서 통상적으로 사용되고 있는 것으로, 예를 들면, 규조토, 소석회 등의 산화물, 인회석 등의 인산염, 석고 등의 황산염, 클레이, 카올린, 벤토나이트, 산성백토, 석영, 실리카 등의 광물질 분말 등의 고체 담체와 충진제, 항응집제, 계면활성제, 유화제, 방부제 등을 추가하여 포함할 수 있다. 또한, 상기 농업용 작물 보호제를 식물에 처리하여 활성성분이 신속방출, 서방출, 지연된 방출할 수 있도록 당업계에서 공지된 방법을 사용하여 제형화할 수 있다. 제형화를 위해서는 통상적으로 사용하는 계면활성제, 희석제, 분산제, 보조제 등의 첨가제를 활성성분과 배합하여 수화제, 현탁제, 유제, 유탁제, 미탁제, 액제, 분산성 액제, 입상수화제, 입제, 분제, 액상수화제, 수면부상성입제, 정제 등 각종 형태로 제제화하여 사용할 수 있다. In the present invention, excipients and diluents contained in the agricultural crop protection agent are those conventionally used in the agricultural field. Examples thereof include oxides such as diatomaceous earth and slaked lime, phosphates such as apatite, sulfates such as gypsum, clay, kaolin, Mineral carriers such as bentonite, acidic clay, quartz, and silica, fillers, anti-coagulants, surfactants, emulsifiers, preservatives and the like. In addition, the agricultural crop protection agent can be formulated into plants using methods known in the art so that the active ingredient can be released rapidly, slowly, or slowly. For the formulation, additives such as surfactants, diluents, dispersants, and adjuvants commonly used may be blended with the active ingredient to prepare a wetting agent, a suspension, an emulsion, a emulsion, a mist, a liquid, a dispersing liquid, a granular wetting agent, A liquid wettable powder, a water-swellable granule, a tablet, and the like.
본 발명에 있어서, 상기 농업용 작물 보호제는 통상의 방법으로 식물에 적용할 수 있다. 식물에 적용함에 있어서는 식물체의 잎, 줄기, 가지, 뿌리, 종자에 직접 처리 살포 또는 도포하거나, 논이나 밭 등의 일반적인 재배 토양 또는 육묘용 상토나 배지에 혼화처리하거나, 또는 물속에서 재배되는 식물체의 경우는 병해를 방제하기 위하여 수면에 처리할 수 있다. 구체적인 적용방법은 도포 처리, 침지 처리, 훈증 처리 또는 살포 처리이며, 예컨대 농업용 작물 보호제를 토양, 식물의 잎, 줄기, 종자, 꽃 또는 열매에 살포할 수 있다. 상기 농업용 작물 보호제를 식물에 적용하기 위하여 물 또는 적절한 매체에 희석하여 사용할 수 있다.In the present invention, the agricultural crop protection agent can be applied to plants by a conventional method. When applied to plants, it may be applied directly to the leaves, stems, branches, roots and seeds of the plants, sprayed or applied to the plants, mixed with the soil or medium used for cultivation in general cultivated soil such as rice fields or fields, The case can be treated on the surface to control the disease. Specific application methods are application treatment, immersion treatment, fumigation treatment or spraying treatment, for example, agricultural crop protection agent can be applied to soil, plant leaf, stem, seed, flower or fruit. The agricultural crop protection agent may be diluted in water or an appropriate medium for application to plants.
또 다른 하나의 양태로서, 본 발명은 상기 배추 무름병 원인균 억제용 항균 조성물을 포함하는 식품 세척제를 제공한다.In another aspect, the present invention provides a food cleansing agent comprising the antimicrobial composition for inhibiting the causative agent of cabbage root rot.
본 발명에 있어서, 상기 식품 세척제는 칠면초(Suaeda japonica) 추출물, 특히 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)을 유효성분으로 포함할 수 있다. 상기 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, n-부탄올 분획물을 유효성분으로 첨가하여 제조한 식품 세척제는 배추 무름병을 유발하는 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)의 생장을 억제하거나 사멸시켜, 배추 무름병 발생을 예방 및 억제/제어할 수 있다.In the present invention, the food liquid is S. japonica (Suaeda japonica) n obtained from the extract, in particular, S. japonica extract ethanol-hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n - butanol fraction (n -butanol fraction) may comprise as the active ingredient. The food cleaner prepared by adding the n - hexane fraction, the chloroform fraction, the ethyl acetate fraction and the n - butanol fraction obtained from the chllycanic ethanol extract as an active ingredient was a Pectobacterium carotovorum subsp. Carotovorum ), thereby preventing, inhibiting / controlling the occurrence of cabbage blight .
본 발명에 있어서, 상기 식품 세척제는 본 발명에 따른 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction) 외에 세척제의 제조 분야에서 일반적으로 사용되는 하나 이상의 부형제 및 첨가제를 포함하여 당 분야의 공지의 방법에 따라 용이하게 제조될 수 있다. 상기 식품 세척제의 경우에는 본 발명에 따른 칠면초 추출물에 식품학적으로 허용되는 용매 또는 희석제를 추가로 첨가하여 적절한 농도로 희석함으로써 제조할 수 있다. 상기 식품 세척제는 배추 무름병 원인균에 의해 무름병이 발생할 수 있는 기주식물, 예컨대, 상추, 무, 배추, 토마토, 고구마, 감자 등의 살균 및 세척에 사용될 수 있다.In the present invention, the food liquid is n obtained from S. japonica ethanol extract according to the invention - the hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction in addition to the n- butanol fraction, one or more excipients and additives commonly used in the manufacture of detergents. In the case of the above-mentioned food cleanser, it can be prepared by additionally adding a pharmaceutically acceptable solvent or diluent to the extract of Chilchomalaceae according to the present invention and diluting it to an appropriate concentration. The food cleaner may be used for disinfection and washing of host plants, for example, lettuce, radish, Chinese cabbage, tomato, sweet potato, potato, etc., which may be blighted by the causative bacteria of cabbage rot.
또 다른 하나의 양태로서, 본 발명은 상기 배추 무름병 원인균 억제용 항균 조성물을 포함하는 식품 첨가제를 제공한다. In yet another aspect, the present invention provides a food additive comprising the antimicrobial composition for inhibiting the causative agent of cabbage acne-causing bacteria.
본 발명에 있어서, 상기 식품 첨가제는 칠면초(Suaeda japonica) 추출물, 특히 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)을 유효성분으로 포함할 수 있다. 상기 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물, 에틸아세테이트 분획물, n-부탄올 분획물을 유효성분으로 첨가하여 제조한 식품 첨가제는 배추 무름병을 유발하는 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)의 생장을 억제하거나 사멸시켜, 배추 무름병 발생을 예방 및 억제/제어할 수 있다.In the present invention, the food additive S. japonica (Suaeda japonica) extract, particularly obtained from an ethanol extract of S. japonica a n-hexane fraction (n -hexane fraction), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction (n - butanol fraction) as an active ingredient. The food additive prepared by adding the n - hexane fraction, ethyl acetate fraction and n - butanol fraction obtained from the chllycolipid ethanol extract as an active ingredient was Pectobacterium carotovorum subsp. Carotovorum ), thereby preventing, inhibiting / controlling the occurrence of cabbage blight .
본 발명에 있어서, 상기 식품 첨가제는 상기 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction) 이외에, 항균 활성을 가지는 다른 공지 항균 물질이나 기타 기능성을 부여할 수 있는 물질을 더 포함할 수 있다. 또한, 식품 첨가제는 상기 칠면초 추출물을 유효성분으로 포함하는 상기 항균 조성물 자체일 수 있으며, 제형화에 필요한 부형제나 희석제 등의 통상적으로 사용되는 첨가제가 더 포함될 수 있다. In the present invention, wherein the food additive is a n obtained from the S. japonica extract ethanol-hexane fraction (n -hexane fraction), ethyl acetate fraction (ethyl acetate fraction), n-butanol addition to the fractions (fraction n -butanol), antibacterial activity Lt; RTI ID = 0.0 > other < / RTI > antimicrobial material or other functional material. In addition, the food additive may be the above-mentioned antimicrobial composition itself containing the above-mentioned chickpea extract as an active ingredient, and may further contain commonly used additives such as excipients and diluents necessary for formulation.
또 다른 하나의 양태로서, 본 발명은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는 상기 항균 조성물을 이용하여 배추 무름병 원인균을 방제하는 방법을 제공한다.In another aspect, the present invention provides a method for controlling cabbage root rot caused by using the above-described antimicrobial composition comprising an extract of Suaeda japonica as an active ingredient.
본 발명의 상기 조성물을 배추 무름병이 발생된 병반 또는 발생이 예상되는 식물 부위, 예컨대, 식물 전체에의 처리, 식물근(root) 처리, 식물 종자 처리, 번식기 중의 식물 조직 처리, 또는 식물 파종이나 식수 전후에 식물 병해 방제용 매체에 포함시켜 처리하는 방법이 포함될 수 있으나, 이에 제한되지는 않는다. The composition of the present invention may be applied to a diseased or diseased lesion in which a Chinese cabbage blight has occurred or to an expected plant part such as a plant as a whole, a root treatment, a plant seed treatment, a plant tissue treatment in a breeding season, And a method of treating the plant disease prevention medium before and after the treatment, but the present invention is not limited thereto.
본 발명에 있어서, 상기 식물은 배추 무름병 원인균에 의해 무름병이 발생할 수 있는 기주식물, 예컨대, 상추, 무, 배추, 토마토, 고구마, 감자 등일 수 있으나, 이에 한정되는 것은 아니다. In the present invention, the plant may be, but not limited to, a host plant in which blight may occur due to a causative organism causing cabbage root rot, such as lettuce, radish, Chinese cabbage, tomato, sweet potato, potato and the like.
본 발명에 있어서, 상기 배추 무름병 원인균은, 이에 제한되지는 않으나, 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)일 수 있다.In the present invention, the causative bacteria of Chinese cabbage root rot, include, but are not limited to, Pectobacterium carotovorum subsp. Carotovorum .
본 발명에서 있어서, 상기 방제 방법은 당업계에 공지된 다양한 방법으로 상기 식물 내지 식물 주변 토양에 처리될 수 있고, 상기 식물 내지 식물 주변 토양에의 처리는 살포, 도포, 관주, 인필트레이션, 분무, 주사, 코팅, 더스팅 또는 침지 등 당업계 공지된 다양한 방법으로 수행될 수 있다. In the present invention, the above-mentioned controlling method can be applied to the soil around the plant or plant by various methods known in the art, and the treatment to the soil around the plant or the plant can be performed by spraying, coating, crossing, infiltration, , Spraying, coating, dusting, or dipping.
또 다른 하나의 양태로서, 본 발명은 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는, 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum) 증식 억제 또는 사멸용 조성물을 제공한다.In another aspect, the present invention relates to a method for producing Pectobacterium, which comprises extract of Suaeda japonica as an active ingredient, carotovorum subsp. Carotovorum ) proliferation inhibiting or killing.
본 발명에 있어서, 상기 칠면초 추출물은 전술한 바와 같다.In the present invention, the chitosan extract is as described above.
본 발명의 일실시예에서, 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)의 살균 효과를 확인하기 위하여, 현탁액 시험법(suspension test; CEN[European Committee for Standardization], EN 1040, 1276)을 수행하여 배추 무름병 원인균인 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 살균 효과를 분석한 결과, 클로로포름 분획물(chloroform fr.) 및 에틸아세테이트 분획물(ethyl acetate fr.)에 의해 처리 20분 후 PCC 균주의 생존율이 완전 제어됨을 확인하였다(도 2).In one embodiment of the invention, a S. japonica n obtained from the ethanol extract of hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction (n -butanol fraction (CEN [European Committee for Standardization], EN 1040, 1276) was carried out in order to confirm the bactericidal effect of Pseudomonas aeruginosa, Pectobacterium carotovorum subsp. Carotovorum , PCC), it was confirmed that the survival rate of the PCC strain was completely controlled after 20 minutes of treatment with chloroform fractions (chloroform fr.) And ethyl acetate fractions (FIG. 2) .
본 발명의 칠면초(Suaeda japonica) 추출물을 유효성분으로 포함하는 상기 조성물은 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum)의 증식 억제 및 사멸 효과가 뛰어나며, 배추 무름병 원인균 방제를 위한 일반적인 화학 농약에 비하여 부작용이 덜할 수 있으므로 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum) 증식 억제 또는 사멸용 조성물로 안전하게 사용될 수 있다.The composition comprising the extract of Suaeda japonica of the present invention as an active ingredient can be obtained by extracting Pectobacterium carotovorum subsp. Carotovorum ), and is superior to general chemical pesticides for the control of pathogenic bacteria caused by Chinese cabbage blight . Therefore , it can be safely used as a composition for inhibiting or preventing the growth of Pectobacterium carotovorum subsp. Carotovorum .
본 발명에 있어서, 상기 펙토박테리움 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum) 증식 억제 또는 사멸용 조성물 중 선택되는 어느 하나는, 실제 포장에서 사용하기 적합한 안정적인 제제화를 목적으로 생화학 농약으로 사용 가능한 계면활성제 또는 증량제 등을 사용하여 수화제, 입상수화제, 액상수화제, 액제, 수용제, 수용성입제 또는 캡슐화제의 형태로 제조될 수 있다.In the present invention, any one selected from the compositions for suppressing or killing Pectobacterium carotovorum subsp. Carotovorum may be used as an interface for use as a biochemical pesticide for the purpose of stable formulation suitable for use in actual packaging A wetting agent, a liquid wettable agent, a liquid agent, a water-soluble agent, a water-soluble granule, or an encapsulating agent using an active agent or an extender.
본 발명의 칠면초 추출물을 유효성분으로 하는 항균 조성물은 천연 약용 염생식물을 원료로 하고, 배추 무름병 원인균인 PCC에 대한 생장 저해 효과, 즉, PCC에 대한 항균 활성 및 살균 효과를 나타내므로, 농작물의 무름병 방제를 위한 소재로 유용하게 활용될 수 있다. 구체적으로, 농작물 수확 후 농산물의 유통 및 저장 중 배추 무름병균에 의한 농작물 부패에 따른 손실을 막을 수 있으며, 농용항생제 대신 사용하여 항생제 사용을 절감하고 내성균 출현 가능성을 감소시키며 환경 안전성 또한 높일 수 있다.The antimicrobial composition comprising the extract of the present invention as an active ingredient exhibits an antimicrobial activity and a bactericidal effect against the growth inhibition effect against PCC which is a causative agent of cabbage acne caused by natural medicinal plants, It can be useful as a material for control. Specifically, it is possible to prevent the loss caused by crop rot caused by the Chinese cabbage root rot during the distribution and storage of the agricultural products after the harvest of the crop, and it can be used instead of the agricultural antibiotic to reduce the use of antibiotics, reduce the possibility of the occurrence of resistant bacteria, and improve the environmental safety.
도 1은 본 발명의 염생식물(halophyte)인 칠면초(Suaeda japonica) 에탄올 추출물의 용매별 분획물을 제조하는 방법을 개략적으로 나타낸 도이다.
도 2는 본 발명의 칠면초 에탄올 추출물의 용매별 분획물의 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 시간별 살균효과를 나타낸 그래프이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram schematically showing a method for producing a solvent-separated fraction of an ethanol extract of Suaeda japonica which is a halophyte of the present invention.
Fig. 2 is a graph showing the effect of the solvent fraction of the extract of chile < RTI ID = 0.0 > cotton < / RTI > extract of the present invention on the pectobacterium carotovorum subsp. Carotoborum carotovorum subsp. Carotovorum , PCC). ≪ / RTI >
이하, 실시예를 통하여 본 발명의 구성 및 효과를 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들 실시예에 의해 한정되는 것은 아니다.Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.
실시예Example 1: One: 칠면초Chili pepper 에탄올 추출물 및 이의 용매 Ethanol extract and its solvent 분획물Fraction 제조 Produce
염생식물(halophyte)인 칠면초(Suaeda japonica)는 온라인을 통하여 구입하였다. 상기 구입한 칠면초 시료는 동결건조한 후 분쇄기를 이용하여 60 메시(mesh)로 분쇄한 후 사용하였다. 칠면초 시료 100g에 70% 에탄올(ethanol) 1ℓ를 첨가한 후 25℃에서 12시간 동안 추출하였다. 추출한 칠면초 시료를 여과 장치를 이용하여 여액만 취하였으며, 이와 같은 방법으로 분리한 잔사에 대하여 동일한 조건으로 2회 더 반복 실시하였다. 이후 0.45 ㎛(Adventec No. 2) 기공 크기의 여과지로 여과하여 얻은 여액을 농축한 후, 동결건조하여 70% 에탄올(ethanol) 조추출물(crude extract)로 사용하였다. 70% 에탄올 조추출물을 물에 녹인 후, 용매의 극성순서에 따라 순차적으로 분획하여 n-헥산(n-hexane), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), n-부탄올(n-butanol), 및 물(water, H2O; aqueous phase) 순서로 총 5개의 용매 분획 층을 얻었다(도 1). 상기 생성된 분획물들은 각각 감압 농축하고, 황산나트륨(sodium sulfate)을 이용하여 수분을 제거한 후, 동결건조하여 사용하였다. 한편, 상기 분획물 제조시에는 EP 등급(extra pure grade; Duksan Co. Ltd.)의 용매와 3차 증류수를 사용하여 수행하였다.The halophyte, Suaeda japonica , was purchased online. The thus-obtained chrysanthemum samples were lyophilized and pulverized to a mesh of 60 mesh using a pulverizer. 1 liter of 70% ethanol was added to 100 g of the sample of Chilpegakchoi, and the mixture was extracted at 25 ° C for 12 hours. The extracts of the extracted chestnut specimens were collected by filtration using a filtration apparatus, and the residues separated in this manner were repeated twice under the same conditions. After filtration with 0.45 ㎛ (Adventec No. 2) pore size filter, the filtrate was concentrated and lyophilized to use as a 70% ethanol crude extract. 70% ethanol and then the crude extract is dissolved in water, and sequentially fractionated by the polarity order of the solvents n - hexane (n -hexane), chloroform (chloroform), ethyl acetate (ethyl acetate), n - butanol (n -butanol) , And water (H 2 O, aqueous phase) (FIG. 1). The resulting fractions were concentrated under reduced pressure, respectively, and sodium sulfate was used to remove moisture, followed by lyophilization. Meanwhile, the fraction was prepared using an EP grade (extra pure grade; Duksan Co. Ltd.) solvent and tertiary distilled water.
실시예Example 2: 2: 칠면초Chili pepper 추출물의 용매별 Solvent of extract 분획물의Fraction 항균 효과 분석 Antibacterial effect analysis
상기 실시예 1에서 수득한 칠면초 에탄올(ethanol) 추출물의 4가지 용매 분획물인 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)의 항균 효과를 확인하기 위하여, 페이퍼 디스크 확산법(paper disc diffusion method)을 이용하여 항균 활성을 측정하였다.S. japonica obtained in Example 1 in ethanol (ethanol) 4 gaji solvent fraction of an extract of n-hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction To determine the antimicrobial activity of the n- butanol fraction, antimicrobial activity was measured using a paper disc diffusion method.
상기 칠면초 추출물의 4가지 용매 분획물 시료의 항균 활성을 측정하기 위해, 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum KACC18645, PCC)은 농촌진흥청으로부터 분양받아 항균 활성 실험에 사용하였다. In order to measure the antimicrobial activity of the four solvent fraction samples of the chylomyrrhizae extract, Pectobacterium carotovorum subsp. Carotovorum KACC18645, PCC) were purchased from RDA and used for the antimicrobial activity test.
구체적으로, 상기 분양받은 PCC 균주는 락토오즈 브로스(Lactose broth; Difco Co., USA)에 접종하여 28℃에서 24시간 배양하였다. 배양한 후, TSA(Tryptic Soy Agar; Difco Co., USA) 멸균 고체 배지 플레이트(petri dish; 90×15㎜, Green Cross Co., Ltd., Korea) 당 6~7 log CFU(colony forming unit)로 도말하고, 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fr.), 클로로포름 분획물(chloroform fr.), 에틸아세테이트 분획물(ethyl acetate fr.), 및 n-부탄올 분획물(n-butanol fr.) 상기 4가지 시료 10㎕를 각각 멸균 디스크-페이퍼(disc-paper; 지름 6.0㎜, Whatsman No.2)에 가하였다. 이때, 처리된 상기 4가지 칠면초 분획물 시료의 최종 농도는 디스크 당 400㎍(400㎍/disc)이었다. 멸균 디스크-페이퍼에 상기 4가지 칠면초 분획물 시료를 각각 가한 후, 28℃에서 24시간 동안 배양하였으며, 배양 후 형성된 생육저지환(clear zone)의 크기를 측정하여 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 항균 활성을 평가하였다. 이때, 항세균제인 앰피실린(ampicillin; Sigma Co., USA)을 디스크 당 100㎍(100㎍/disc)의 농도로 처리한 군을 대조군으로는 사용하였다. 한편, 생육저지환의 크기는 육안으로 생육이 나타나지 않는 부분의 지름을 ㎜ 단위로 측정하였으며, 3회 이상 평가 후 평균값으로 나타내었다.Specifically, the pre-cultured PCC strain was inoculated into lactose broth (Difco Co., USA) and cultured at 28 ° C for 24 hours. After culturing, 6 to 7 log CFU (colony forming unit) per TSA (Tryptic Soy Agar; Difco Co., USA) sterilized solid medium plate (petri dish; 90 x 15 mm, Green Cross Co., a smear in and obtained from S. japonica ethanol extract n - hexane fraction, chloroform fraction, ethyl acetate fraction, and n (n -hexane fr.) ( chloroform fr.) (ethyl acetate fr.) - butanol fraction (fr n -butanol ). 10 [mu] l of the above four samples were added to each sterilized disc-paper (diameter 6.0 mm, Whatsman No. 2). At this time, the final concentration of each of the four fractions obtained from the above-mentioned fractions was 400 μg / disc (400 μg / disc). Samples of the fractions of the four fractions were added to the sterilized disk-paper and incubated at 28 ° C for 24 hours. The size of the clear zone formed after the culture was measured to determine the size of the pectobacterium carotenoid Pectobacterium < RTI ID = 0.0 > carotovorum subsp. Carotovorum , PCC). At this time, ampicillin (Sigma Co., USA), an anti-bacterial agent, was treated at a concentration of 100 μg per disc (100 μg / disc) as a control. On the other hand, the size of the growth inhibition ring was measured in terms of the diameter of the portion where the growth did not appear on the naked eye in mm, and the average value was evaluated after 3 or more evaluations.
칠면초 에탄올 추출물의 4가지 용매 분획물인 n-헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물, 및 n-부탄올 분획물의 PCC 균주에 대한 항균 활성을 측정한 결과, 하기 표 1에 나타낸 바와 같이, 상기 4가지 분획물 모두 PCC 균주에 대하여 생장 저해 활성이 우수함을 확인하였다.The antibacterial activities of n - hexane fraction, chloroform fraction, ethylacetate fraction and n - butanol fraction of the four solvent fractions of the ethanol extract of chile peach were measured. As shown in the following Table 1, All of the PCC strains were confirmed to have excellent growth inhibitory activity.
한편, 상기 4가지 용매 분획물은 PCC 균주에 대한 생장 저해 활성에 있어 분획물 간 다소 차이를 나타내었는데, 에틸아세테이트 분획물 및 n-헥산 분획물, n-부탄올 분획물, 클로로포름 분획물 순(Ethyl acetate fr. = n-Hexane fr. > n-Butanol fr. > Chloroform fr.)으로 생장 저해 활성이 나타났다. 이에 반해, 항세균제로 알려진 대조군으로 사용한 앰피실린(ampicilin)은 PCC 균주에 대한 생장 저해 활성을 전혀 보이지 않았다.On the other hand, the four kinds of solvent fraction is in the growth-inhibiting activity of the PCC strain eotneunde indicate little difference between the fraction, ethyl acetate fraction and n - hexane fraction, n - butanol fraction, chloroform fraction net (Ethyl acetate fr = n. - Hexane fr. ≫ n- Butanol fr. ≫ Chloroform fr.). On the other hand, ampicilin used as a control group known as an antibacterial agent showed no growth inhibitory activity against the PCC strain.
실시예Example 3: 3: 칠면초Chili pepper 추출물의 용매별 Solvent of extract 분획물의Fraction 살균 효과 분석 Analysis of bactericidal effect
상기 실시예 1에서 수득한 칠면초 에탄올(ethanol) 추출물의 4가지 용매 분획물인 n-헥산 분획물(n-hexane fraction), 클로로포름 분획물(chloroform fraction), 에틸아세테이트 분획물(ethyl acetate fraction), n-부탄올 분획물(n-butanol fraction)의 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)에 대한 살균 효과를 확인하기 위하여, 현탁액 시험법(suspension test; CEN[European Committee for Standardization], EN 1040, 1276)을 일부 변형하여 실시하였다.S. japonica obtained in Example 1 in ethanol (ethanol) 4 gaji solvent fraction of an extract of n-hexane fraction (n -hexane fraction), chloroform fraction (fraction chloroform), ethyl acetate fraction (ethyl acetate fraction), n-butanol fraction (CEN [European] test to determine the bactericidal effect of Pectobacterium carotovorum subsp. Carotovorum (PCC), a causative organism of cabbage root rot of n- butanol fraction, Committee for Standardization], EN 1040, 1276).
구체적으로, PCC 균주의 초기 균수를 4 log CFU(colony forming unit)/㎖로 조정하여 준비한 후, 칠면초 에탄올 추출물로부터 수득한 n-헥산 분획물(n-hexane fr.), 클로로포름 분획물(chloroform fr.), 에틸아세테이트 분획물(ethyl acetate fr.), 및 n-부탄올 분획물(n-butanol fr.) 상기 4가지 시료의 최종 농도가 200㎍/㎖이 되도록 각각 처리하였다. 처리한 후, 28℃에서 0~120분 동안 배양하면서 처리 시간별 PCC 균주의 생존율을 확인하였다. 이때, PCC 균주의 생존율은 TSA(Tryptic Soy Agar; Difco Co., USA) 멸균 고체 배지에 상기 처리 시료를 0분, 10분, 20분, 30분, 60분, 90분 및 120분 시간별로 취하여 도말하고, 28℃에서 24시간 동안 배양하여 생균수를 측정하여 Log10 CFU/㎖ 값으로 나타내었다.More specifically, the initial number of bacteria of the
칠면초 에탄올(ethanol) 추출물의 4가지 용매 분획물에 따른 배추 무름병 원인균인 펙토박테리움 카로토보룸 아종 카로토보룸(Pectobacterium carotovorum subsp. Carotovorum, PCC)의 생존율을 확인한 결과, 하기 도 2에 나타낸 바와 같이, 칠면초 에탄올 추출물의 각각의 용매 분획물에 따른 PCC 균주 억제 시간 및 살균 효과가 각 분획물 간에 차이가 있음을 확인하였다.The four solvent fractions of the ethanol extract of chile peach were used to isolate Pectobacterium, a causative organism of Chinese cabbage root rot, carotovorum subsp. Carotovorum , PCC). As shown in FIG. 2, it was confirmed that the inhibition time and the bactericidal effect of PCC strain on the fraction of the solvent of the ethanol extract of chile peach were different among the fractions.
구체적으로, 클로로포름 분획물(chloroform fr.) 및 에틸아세테이트 분획물(ethyl acetate fr.)의 경우, 처리 20분 후 PCC 균주의 생장을 4 log 이상 완전히 제어함을 확인하였으며, n-부탄올 분획물(n-butanol fr.)의 경우에는, 처리 120분 후 PCC 균주의 생장을 4 log 이상 완전히 제어함을 확인하였다. 이에 반해, 상대적으로 n-헥산 분획물(n-hexane fr.)의 경우, 처리 120분 후에도 PCC 균주의 생장을 완전히 제어하지는 못함을 확인하였다.Specifically, the chloroform fraction and the ethyl acetate fraction of the time, 20 minutes after treatment was confirmed that the growth of the complete control of more than 4 log of PCC strain, n (chloroform fr.) ( Ethyl acetate fr.) - butanol fraction (n -butanol fr.), it was confirmed that the growth of the PCC strain was completely controlled by 4 log or more after 120 minutes of treatment. On the other hand, a relatively n - it was confirmed that the hexane fraction of the case, the process does not completely control the growth of strain PCC even after 120 minutes (n -hexane fr.).
Claims (9)
An antimicrobial composition for inhibiting root rot caused by a Chinese cabbage ( Suaeda japonica ) as an active ingredient.
The antimicrobial composition according to claim 1, wherein the extract is an organic solvent extract selected from the group consisting of ethanol, hexane, chloroform, ethyl acetate and butanol.
The antimicrobial composition according to claim 1, wherein the extract is a fraction obtained by adding hexane, chloroform, ethyl acetate or butanol to the ethanol extract derived from the chili pepper obtained by adding ethanol to the chili pepper.
The antimicrobial composition according to claim 1, wherein the causative agent of Chinese cabbage rot is Pectobacterium carotovorum subsp. Carotovorum .
An agricultural crop protection agent comprising the antimicrobial composition for inhibiting root rot caused by the Chinese cabbage according to any one of claims 1 to 4.
The food cleaner according to any one of claims 1 to 4, which comprises the antimicrobial composition for inhibiting the causative bacteria of Chinese cabbage rot.
The food additive according to any one of claims 1 to 4, which comprises the antimicrobial composition for inhibiting root rot caused by Chinese cabbage.
A method for controlling cabbage root rot caused by the composition according to claim 1.
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| KR20200082023A (en) | 2018-12-28 | 2020-07-08 | 주식회사 비오엘연구소 | Anti-microbial coating composition |
| KR20220038215A (en) | 2020-09-18 | 2022-03-28 | 한국식품연구원 | Sterilization method for controlling Pectobacterium carotovorum subsp. carotovorum using weak acidic hypochlorous acid water |
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