Aspergillus terreus로부터 항진균성 물질의 분리 및 구조분석

김근기,박기훈,문석식,강규영 ( Keun Ki kim,Ki Hun Park,Suk sik Moon,Kyu Young Kang ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.6

In the course of search antagonistic fungi from soil in green house, four kind of fungi (AF1, AF2, AF3, AF4) were isolated, which have activities aganist Phytophthora capsici, Botrytis cinera, Rhizoctomia solani, Pythium ultimum and Fusarium oxysporum. The AF2 was identified according to the morphological description of Aspergillus terreus. This antagonistic fungus inhibiting various plant pathogens was effective to reduce disease incidence of cucumber seedlings caused by mixed inoculum of Rhizoctonia solani, Pythium ultimum and Fusarium oxysporum. Antifungal compound I was isolated and purified by fresh chromatography from A. terreus. The ¹H and ^(13)C assignment of compound I was achieved from two-dimensional ¹H-¹H COSY, HMQC, HMBC with the add of homonuclear and heteronuclear double resonance experiment. The compound I was identified butyrolactone I (α-oxo-β-(p-hydroxyphenyl)-γ-(p-hydroxy-m-3,3-dimethyl-allylbenzyl)-γ-methoxy-carbonyl-γ-butyrolactone, C_(24)H_(24)O_7, M.W.=424).

초피첨가 전통장류의 항균 및 항암활성

김근기,박현철,손홍주,김용균,이상몽,최인수,최영환,신택순,Kim, Keun-Ki,Park, Hyean-Cheal,Son, Hong-Joo,Kim, Yong-Gyun,Lee, Sang-Mong,Choi, In-Soo,Choi, Young-Whan,Shin, Teak-Soon 한국생명과학회 2007 생명과학회지 Vol.17 No.8

초피나무(Z. piperitum) 열매는 다양한 생리활성을 갖고 있으며, 과피 용매추출물은 세균에 대한 강한 항균활성을 갖고 있어, 간장과 된장제조에 초피를 첨가하여 생리활성효과를 조사하였다. 초피를 간장제조에 1%, 2%, 4% 첨가하여 제조하고, 간장추출물을 식중독균(Sta. aureus, Sal. typhimurium, V. parahemolyticus, E. coli 0157:H7)에 대한 항균활성을 조사했다. 초피를 2% 첨가한 간장은 E. coli 0157:H7과 V. parahemolyticuS 의 생육을 각각 70%와 50% 억제했으며, Sal. typhimurium 과 Sta. aureus 에 대한 항균효과는 농도와 간장제조시기에 따라 40% ${\sim}$ 60% 의 생육을 억제했다. 초피된장의 용매추출물에서는 약한 항균활성을 확인할 수 있었다. 초피간장의 추출물을 백혈병세포 U-937에 처리하여 caspase-3 활성으로 항암활성을 조사했다. 그 결과 초피의 량을 1%, 2%, 4% 첨가한 간장의 추출물은 초피를 침가하지 않은 간장보다 caspase-3 활성이 각각 4배, 12배, 15배 활성이 증가하는 것이 확인되어졌다. 이는 초피간장의 추출물이 apoptosis를 유도하는 것을 의미하며, 된장의 용매추출물은 caspase-3 활성에 아무런 영향이 없었다. 초피간장 추출물을 처리한 세포의 죽음이 necrosis인지 apoptosis인지 알아보기 위하여 세포내의 DNA fragmentation을 확인한 결과, 초피간장 추출물을 처리한 세포는 DNA 단편화가 형성되어졌다. 초피틀 1% 첨가한 것에서는 fragmentation을 확인하기 힘들었고, 2%와 4%초피를 첨가한 간장추출물은 DNA fragmentation을 강하게 일으켰다. 초피를 첨가한 간장추출물이 caspase를 활성화시키고, DNA fragmentation을 일으키는 apoptosis 기작에 의해 암세포를 죽음으로 유도하는 초피간장의 항암효과를 입증했다. The fruits of Zanthoxylum piperitum are known as having various physiology vitality, and the abstraction ingredient of the pericarp is also known as having strong antibiotic activities against various bacteria. Therefore, this study was carried out to estimate the effect of physiology vitality when the abstraction ingredient of Z. piperitum was added in soy sauce(Chopi-kanjang) and soybean paste(Chopi-doenjang). For the antibiotic activity against the pathogens of sitotoxism such as Staphylococcus aureus, Salmonella typhimurium, Vibrio parahemolyticus, Escherichia coli 0157:H7, the extracts of the Chopi-kanjang was added 1%, 2%, 4% pericarp of Z. piperitum in the manufacturing process of soy source. According to the results, the growth of E. coli 0157:H7 and V. parahemolyticus were respectively inhibited as 70% and 50% by the Chopi-kanjang added 2% of the ingredient. For the antibiotic effects of the aforementioned Chopi-kanjang against Sal. typhimurium and Sta. aureus, the growth of those pathogens was also inhibited between 40% and 60% according to the manufacturing period of Chopi-kanjang. It was confirmed that the antibiotic activity using the mixture of the abstraction ingredient and Chopi-doenjang was lower than those of Chopi-kanjang. In order to estimate the anticancer activity using by caspase-3 activity, the mixture of the abstraction ingredient of the pericarp of Z. piperitum and Chopi-kanjang was treated to leukemia cells. According to the results, the activities of caspase-3 using the mixture added 1%, 2% and 4% of the abstraction ingredient were respectively increased as much as 4, 12, 15 times comparing with the control which was treated with the soy source only. It could be that the mixture of the abstraction ingredient of the pericarp of Z. piperitum and soy source induced apoptosis, and the mixture of the abstraction ingredient and soybean paste had no effect on the activity of caspase-3. In order to find out the death of the aforementioned cells caused by necrosis or apoptosis, DNA fragmentation in the cell was examined. U-937 cells showed apoptotic DNA fragmentation in the incubation with Chopi-kanjang extract.

Stevia rebaudiana Bertoni의 Steviol 생합성 효소 ent-Kaurenoic Acid 13-Hydroxylase의 특성

김근기,Hitoshi Shibata ( Keun Ki Kim ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.6

Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. [^(14)C]-methylated ent-KA was used to localize ent-KA hydroxylase. [^(14)C]-methyl-KA was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in strotna fraction of either Spinacia deracea and Solidago altissima. The reaction products using [^(14)C]-methyl-KA were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

Aspergillus terreus로부터 항진균성 물질의 분리 및 구조분석

김근기,박기훈,문석식,강규영,Kim, Keun-Ki,Park, Ki-Hun,Moon, Suk-Sik,Kang, Kyu-Young 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.44 No.4

In the course of search antagonistic fungi from soil in green house, four kind of fungi (AF1, AF2, AF3, AF4) were isolated, which have activities against Phytophthora capsici, Botrytis cinera, Rhizoctonia solani, Pythium ultimum and Fusarium oxysporum. The AF2 was identified according to the morphological description of Aspergillus terreus. This antagonistic fungus inhibiting various plant pathogens was effective to reduce disease incidence of cucumber seedlings caused by mixed inoculum of Rhizoctonia solani, Pythium ultimum and Fusarium oxysporum. Antifungal compound I was isolated and purified by fresh chromatography from A. terreus. The $^1H$ and $^{13}C$ assignment of compound I was achieved from two-dimensional $^1H-^1H\;COSY$, HMQC, HMBC with the add of homonuclear and heteronuclear double resonance experiment. The compound I was identified butyrolactone I (${\alpha}$-oxo-${\beta}$-(p-hydroxyphenyl)-${\gamma}$-(p-hydroxy-m-3,3-dimethyl-allylbenzyl)-${\gamma}$-methoxycarbonyl-${\gamma}$-butyrolactone, $C_{24}H_{24}O_7$, M.W.=424). 시설내 토양으로부터 식물병원균에 활성을 갖는 길항균을 탐색하는 과정에서 주요 식물 병원균인 Phytophthora capsici, Botrytis cinerea, Rhizoctonia solani, Pythium ultimum 및 Fusarium oxysporum에 활성을 갖는 4종류의 길항균(AF1, AE2, AE3, AF4)을 분리하였다. 이들 중 가장 높은 활성을 나타낸 AE2균을 동정한 결과, 생육한 균은 짙은 오렌지색과 황갈색을 띄었으며, 현미경의 morphology는 column형의 conidial head와 foot cell로 부터 곧게 뻗은 분생자와 2중층의 포자낭병을 형성했다. 그리고 MY20 agar 배지에서 투명한 둥근세포의 형성등이 Aspergillus terreus의 형태와 일치 하였다. Pot에서 병원균과 A. terreus를 동시에 처리하여 오이의 생육을 조사한 결과, 토양중량당 A. terreus의 균체를 1% 처리에 40%, 5% 처리에 57%, 10% 처리에는 75% 병발생율을 억제시켰다. 그리고 A. terreus의 배양여액으로부터 activity-guided fractionation을 실시하여 항진균성 물질인 화합물 I 을 얻었다. $^1H$, $^{13}C\;NMR$, DEPT, $^1H-^1H\;COSY$, HMQC, HMBC 및 질량스텍트럼을 분석한 결과 화합물 I 은 butyrolactone I (${\alpha}$-oxo-${\beta}$-(p-hydroxyphenyl)-${\gamma}$-(p-hydroxy-m-3,3-dimethyl-allylbenzyl)-${\gamma}$-methoxycarbonyl-${\gamma}$-butyrolactone, $C_{24}H_{24}O_7$, M.W. = 424)로 동정되었다.

적응 뉴로-퍼지 추론 시스템을 이용한 스윙-업 도립진자 제어

김근기(Kim Keun-Ki),유창완,홍대승,신자호,최창호,최용길,송영목,임화영(Yim Wha-Yeong) 대한전기학회 2001 대한전기학회 학술대회 논문집 Vol.2001 No.7

한국제조기업의 기술협력 파트너쉽 유형과 제품혁신 성과의 통계적 상관관계에 대한 연구

김근기(Keun-Ki Kim),이동희(Dong-Hee Lee) 대한산업공학회 2020 대한산업공학회지 Vol.46 No.5

Characterization of an antifungal compound isolated from an antagonistic fungus Aspergillus terreus against phytopathogenic fungi

Keun-Ki Kim(김근기),Jae-Gon Kang(강재곤),Yong-Lark Choi(최용락),Han-Dae Yun(윤한대),Ho-Sung Ha(하호성),Kyu-Young Kang(강규영) 한국농약과학회 1998 농약과학회지 Vol.2 No.1

Bacillus sp. AB02를 이용한 식물 병원균에 대한 생물검정

김근기(Keun Ki Kim),김용철(Yong Chul Kim),최영환(Young Whan Choi),신택순(Taek soon Shin),박기도(Ki Do Park),강위금(Ui Gum Kang),최용락(Yong Lark Choi),박현철(Hyean Cheal Park) 한국생명과학회 2008 생명과학회지 Vol.18 No.6

겨울철 과채류 시설 재배 단지에서는 저온 다습하여 균핵병 발생이 많아 피해가 심각하기 때문에, 미생물을 이용한 생물학적 방제법으로 균핵병을 방제하고자 길항균을 분리하였다. 시설 재배 토양으로부터 길항세균 55종을 분리하였으며, 분리한 길항세균 중에 가장 활성이 뛰어난 균주를 AB02 균주로 명명하고 이를 이용하여 식물병원균 5종에 대한 항균 스펙트럼을 조사하였다. 그 결과 잿빛곰팡이병원균인 Botrytis cinerea와 균핵병원균인 Sclerotinia sclerotiorum에 항균효과가 뛰어났다. AB02 균주의 균핵병에 대한 생물학적 방제실험은을 위해 들깨 pot 실험으로 병발생 억제효과와 병 방제효과를 조사했다. 그 결과 균핵병 발생 억제효과는 40%로 나타났고, 병 방제효과는 62%로 아주 높게 나타났다. AB02 균주의 식물생육 촉진 효과는 전장이 120%, 생체중량이 141% 증가되었으며, 엽수와 엽면적은 각각 121%와 185%의 생육 촉진 효과를 보였다. 그리고 뿌리 생육조사에서는 생체 중량이 무처리보다 327% 증가하였다. 항균활성과 식물 생육 촉진 활성을 갖는 AB02 균주를 동정한 결과, Bacillus sp.로 확인되었다. In the greenhouse fields for fruits and vegetables during the winter in Korea, there are serious damages by the sclerotium diseases due to the low temperature and humidity. This study was carried out to select an antagonic agent for the biological control of the sclerotium diseases. The 55 antagonic agents were selected from the rhizosphere in soil where the fruits and vegetables were cultivated in the green house fields, and strain AB02 among the tested isolates was estimated to be the strongest antagonist against the sclerotium disease. Using strain AB02, the antifungal spectrum was tested against 5 different plant pathogens. According to the results of the test, strain AB02 showed the high antagonistic effect against Botrytis cinerea and Sclerotinia sclerotiorum. For the experiment of biological control against the sclerotium disease, it was estimated the suppression effect and the control effect by the strain in the pot experiment using the green perilla. According to the result of the pot experiments, the suppression effect was 40% and the control effect was 62%, respectively. For the stimulation effect of the tested plant growth by strain AB02 compared to the control, it was improved as 120% for the total length, 141% for the liveweight, 121% for the total number of leaves, 185% for the leaf area, and 327% for the liveweight of the root, respectively. Strain AB02 showing the antagonistic effect against the sclerotium disease and the stimulation effect for the plant growth was identified as Bacillus sp.

미생물,발효,생물공학 : Burkhoderia sp. AK-17에 의한 잎들깨 재배의 생물학적 조절

김근기 ( Keun Ki Kim ),김용균 ( Yong Kyun Kim ),손홍주 ( Hong Joo Son ),최영환 ( Young Whan Choi ),강규영 ( Kyu Young Kang ) 한국응용생명화학회 2005 Applied Biological Chemistry (Appl Biol Chem) Vol.48 No.1

중금속 오염 농경지 토양에서 바닥재 시용에 의한 카드뮴 식물이용성 저감효과

김성은 ( Sung Un Kim ),김용균 ( Yong Gyun Kim ),이상몽 ( Sang Mong Lee ),박현철 ( Hyean Cheal Park ),김근기 ( Keun Ki Kim ),손홍주 ( Hong Joo Son ),윤성욱 ( Sung Wook Yun ),김상윤 ( Sang Yoon Kim ),홍창오 ( Chang Oh Hong ) 한국환경농학회 2016 한국환경농학회지 Vol.35 No.2

바닥재에 의한 토양 내 카드뮴의 부동화기작 구명 및 바닥재 시용에 의한 상추의 카드뮴 흡수량 저감 효과를 확인하고자 카드뮴으로 오염된 토양에 바닥재를 0, 20, 40, 80 kg/ha로 처리 한 후 상추 내 카드뮴 흡수특성과 토양 내 카드뮴의 특성을 조사하였다. 바닥재 시용량 증가에 따른 pH 및 음하전도 값의 통계적 유의차는 없었으나 바닥재 시용량이 증가할수록 토양의 pH 및 음하전도가 증가하였다. 바닥재 처리에 따른 토양 내 카드뮴의 분획특성을 조사한 결과 수용성 카드뮴의 경우 바닥재 처리량이 증가 할수록 토양 내 수용성, 치환성 및 carbonate 결합태, 유기물 결합태 형태의 카드뮴 함량이 감소되었으며, 특히 Fe/Mn 결합태의 카드뮴의 경우 바닥재 처리량이 증가 할수록 유의하게 감소되어졌다. 반면, 바닥재의 처리량이 증가할수록 광물내 고정태 카드뮴의 경우 유의하게 증가하였다. 상추의 재배기간 동안 가시적인 독성현상 및 생육저해 현상은 나타나지 않았다. 바닥재의 처리량이 증가할수록 상추 내 카드뮴의 농도는 감소하였다. 이는 토양내 카드뮴 분획특성에서 나타난 것처럼 바닥재 시용량이 증가 할수록 식물이 이용 가능한 카드뮴의 형태가 감소됨에 따라 상추내 캄드뮴의 농도 역시 감소되었다고 판단된다. BACKGROUND: Since bottom ash (BA) contains considerable amounts of CaO and MgO, it could be a useful amendment to increase soil pH and to immobilize cadmium (Cd). This study was conducted to evaluate effect of BA application in reducing Cd phytoavailability. METHODS AND RESULTS: Bottom ash was applied at the rate of 0, 20, 40, and 80 Mg/ha to Cd contaminated soil, and then lettuce was cultivated under field condition. soil pH and net negative charge increased slightly with increasing BA application; however, there was no statistical difference among the rates. Water soluble, exchangeable+acidic, reducible, and oxidizable fraction of Cd decreased with increasing bottom ash application rate, whereas residual fraction of Cd increased with increasing bottom ash application rate. Lettuce yield increased with rate of bottom ash up to 40 kg/ha. Visual evidences of cadmium toxicity and growth inhibition were not found during lettuce cultivation. CONCLUSION: Bottom ash was effective to reduce phytoextractability of Cd and to increase lettuce yield. Conclusively, BA could be a good soil amendment to reduce Cd phytoavailability in contaminated arable soil.