Rhabditidae과 선충의 COⅡ 유전자 클로닝 및 염기서열 분석

이상몽 ( Sang Mong Lee ),손홍주 ( Hong Joo Son ),김근기 ( Keun Ki Kim ),홍창오 ( Chang Oh Hong ),박현철 ( Hyean Cheal Park ) 한국환경과학회 2019 한국환경과학회지 Vol.28 No.1

Cytochrome c oxidase subunit II gene(CO gene) is subunit of cytochrome oxidase, which is complex IV of mitochondria electron transport system. It has been frequently used in molecular phylogenetic studies because the speed of its DNA variation is faster than that of nucleus. It is especially useful in phylogenetic study of molecular biology in insects. In this study, we cloned and sequenced CO gene of mitochondria DNA from Rhabditidae family nematode. Our results showed that this gene is comprised of 696 base pairs(bp). In the analysis of similarity of this gene with other known genes of 14 species of nematodes in Rhabditida order, we identified that this gene has high similarity with that of Caenorhabditis briggsae(86.0%) and C. elegans(85.6%) in Rhabditidae family. On the meanwhile, it has very low similarity with that of Angiostrongylus cantonensis(31.8%) in Angiostrongylidae family and Metastrongylus salmi(31.6%) in Metastrongylidae family. Based on the results of this study, we suggest that this nematode is closely related with that of Caenorhabditis genus in Rhabditidae family.

한국에서 개발된 곤충유래 약용버섯인 누에동충하초의 생산기술개발 및 약리학적 특성

이상몽(Sang Mong Lee),김용균(Yong Gyun Kim),박현철(Hyean Cheal Park),김근기(Keun Ki Kim),손홍주(Hong Joo Son),홍창오(Chang Oh Hong),박남숙(Nam Sook Park) 한국생명과학회 2017 생명과학회지 Vol.27 No.2

동충하초(Cordyceps)는 B.C. 2,000년 이후로부터 지금까지 중국, 한국, 일본 등 아시아 국가에서 잘 알려진 전통적인 불로장생의 비약으로 사용되어져 왔다. 곤충기생 곰팡이균으로서 원조 동충하초로 불리는 Cordyceps속에 속하는 Cordyceps sinensis는 우리나라 한반도에서는 발견되지 않고 있다. 그 이유는 해당 곰팡이균에 특정적인 기주곤충인 박쥐나방(Hepialus armonicanus)이 우리나라에는 존재하지 않기 때문으로 보인다. 따라서 차선책으로서 한국형의 동충하초, 즉 Paecilomyces tenuipes 동충하초균에 대한 곤충생체배지로서 누에(Silkworm, Bombyx mori)를 기주곤충으로 사용하는 누에동충하초의 인공적인 생산방법을 세계 최초로 개발하게 된 것이다. 따라서 본 총설은 누에동충하초를 개발하게 된 역사적 배경과 생산방법 및 약리학적 효능과, 비곤충유래 동충하초인 인공배지 배양동충하초(밀리타리스동충하초: Cordyceps militaris, 붉은자루동충하초: Cordyceps pruinosa 등)의 배양조건과 동충하초 품종육성, 배지배양조건, 동충하초의 식품에의 응용 가능성, 화학적 구성성분 및 약리적 효능 등에 대해 고찰하였다. 곤충유래의 누에동충하초 및 비곤충유래동충하초균사체 및 자실체동충하초의 주요 약리작용은 항종양, 면역증강, 항피로, 항스트레스, 항산화, 항노화, 항당뇨, 항염증, 항혈전, 항지질 및 해충방제 효능으로 요약된다. 동충하초의 주요 생리활성물질들은 단백다당체(헥소스, 헥소사민), 코디세핀, 만니톨, 산성다당체 등이다. 단백다당체와 부타놀 추출물은 매우 강한 항종양 효과를 보이나 세포독성은 나타내지 않았다. 만니톨은 종양유발쥐의 수명을 유의성 있게 연장하였다. 에르고스테롤은 항종양 활성은 보이지 않았으나 왕성한 식세포활성을 보였다. 따라서 우리가 높은 관심을 가지고 있는 누에동충하초의 항종양 활성은 세포독성효과보다는 면역력증강효과에 기인하는 것으로 사료된다[164]. Cordyceps is a traditional Chinese medicinal herb well-known in China, Korea and Japan since B.C. 2,000. The original entomopathogenic fungus, Cordyceps sinensis belonging to the genus Cordyceps could not be found inside Korean peninsula due to the absence of the host insect for the corresponding entomogenous fungus. The development of artificial production methods of Korean type Cordyceps using the silkworm Bombyx mori as in vivo culture medium for the the entomopathogenic fungus Paecilomyces tenuipes is the first, and wonderful occasion in the research history of insect industry of this global world. The aim of this article is to review the historical research background, mass-production methods, and pharmacological effects of the silkworm-dongchunghacho (Paecilomyces tenuipes) which is a newly developed Korean medicinal insect-borne mushroom, and another non-insect-borne medicinal mushroom (Cordyceps militaris and Cordyceps pruinosa). Their biological actions include anti-tumor, immunostimulating, anti-fatigue, anti-stress, anti-oxidant, anti-aging, anti-diabetic, anti-inflammatory, anti-thrombosis, hypolipidaemic and insecticidal effects. The bioactive principles are protein-bound polysaccharides (hexose, hexosamin), cordycepin, D-manitol, acidic polysaccharide etc. Protein-bound polysaccharides and n-butanol fractions were demonstrated to show a significant anti-tumor activities but did not show a cytotoxicities. D-mannitol exhibited a significant prolongation of the life span in tumor bearing mice. Ergosterol did not show an efficient anti-tumor activity, but showed a significant phagocytosis enhancing activity. Anti-tumor activity of silkworm-dongchunghacho might be attributed to immuno-stimulating activities rather than cytotoxic effects [164]. Also this review comprises the breeding of Dongchunghacho varieties, optimization of culture conditions, improvement of learning and memory by Dongchunghacho, application of them as foods and chemical constituents.

cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi

Sang Mong Lee(이상몽),Jae Sam Hwang(황재삼),Nam Sook Park(박남숙),Yong Gyun Kim(김용균),Keun Ki Kim(김근기),Hong Joo Son(손홍주),Hyun Chul Park(박현철),Byung Rae Jin(진병래) 한국생명과학회 2010 생명과학회지 Vol.20 No.8

상수리 잎을 먹고 자라는 야생견사곤충의 일종인 작잠의 저장단백질인 아릴포린 유전자의 cDNA를 클로닝하고 발육경과에 따른 유전자발현의 양상을 조사 검토하였다. 작잠의 아릴포린 유전자의 cDNA는 2,112 bp의 ORF(open reading frame)를 포함하여 2,234 bp임을 밝혔다. 작잠 아릴포린 유전자의 cDNA염기서열로부터 아미노산 서열을 검토한 결과 방향족 아미노산인 페닐알라닌(phenylalanine)과 티로신(tyrosine)의 성분이 높은 아미노산 서열구조를 보였으며 ORF로부터 계산한 단백질의 분자량은 83,439 Da 이었다.작잠 아릴포린 저장단백질의 아미노산서열을 다른 곤충의 서열과 그 상동성을 비교 분석한 결과 세크로피아잠(H. cecropia)과는 78%, 담배나방의 알파단량체(M. sexta- α subunit)와는 71%, 담배나방의 베타단량체(M. sexta β-subunit)와는 62% 그리고 가잠(B. mori)과는 64%의 아미노산서열 상동성을 각각 보였다. 또, Northern blot analysis에서 유충 5령기의 중장, 중부실샘, 후부 실샘에서는 아릴포린 유전자가 발현되지 않고 오로지 지방체 조직에서만 아릴포린 유전자가 발현되었음을 확인하였다. 아릴포린 유전자는 특히 종령인 5령 유충의 초기에 발현강도가 높았으며 중, 후기로 갈수록 그 강도가 감소하였다. 하지만 번데기시기에는 흔적 정도의 해당 mRNA가 검출되었으며, 이와 같이 검출된 mRNA는 불활성화된 것으로 추정된다. 이상의 결과에서 작잠 아릴포린 유전자의 cDNA가 클로닝되었으며, 이 유전자는 유충기특이적 발현 및 지방체 조직특이적 발현양상을 보인 점이 본 연구에서 확인되었다. The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly A<SUP>+</SUP> tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta α subunit, M. sexta β subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

환경친화적 미생물농약으로서의 잠재성을 가진 세균의 분리 및 특성

이예람(Ye-Ram Lee),이상미(Sang-Mee Lee),장은영(Eun-Young Jang),홍창오(Chang-Oh Hong),김근기(Keun-Ki Kim),박현철(Hyean-Cheal Park),이상몽(Sang-Mong Lee),김용균(Young-Gyun Kim),손홍주(Hong-Joo Son) 한국생명과학회 2015 생명과학회지 Vol.25 No.12

화학농약의 오남용은 환경오염을 유발하고, 사람의 건강에 악영향을 미치므로 그 대안으로 미생물을 이용한 생물학적 방제에 대한 관심이 높아지고 있다. 본 연구에서는 환경에서 안정성을 유지할 수 있는 생물방제균을 확보하기 위하여 잿빛곰팡이병균인 Botrytis cinerea의 생육을 억제할 수 있는 Bacillus 속 균주를 탐색한 후, 선정된 균주의 항진균 물질 생산을 위한 조건, 항진균 기작 및 물리화학적 안정성에 대하여 조사하였다. 가장 높은 항진균 활성을 가진 균주는 고추의 근권 토양에서 분리된 AF4 균주였으며, 표현형적 특성 및 16S rRNA 유전자 염기서열에 근거하여 동정한 결과, Bacillus subtilis로 확인되었다. AF4 균주는 최적 탄소원으로 glycerol을, 최적 질소원으로 casein을 요구하였으며, 25-40℃ 및 pH 7-10에서 항진균 활성을 나타내었다. 최적조건에서 항진균 활성은 140±3 AU/ml로, 기본배지보다 활성이 증가했음을 알 수 있었다. AF4 균주는 B. cinereas 균사를 형태적으로 변화시킴으로써 항진균 효과를 나타내는 것으로 추정되었으며, 다양한 식물병원성 곰팡이의 생육을 억제할 수 있었다. 또한 AF4 균주가 생성하는 항진균 물질은 열, pH, 유기용매 및 단백질 분해효소 처리에도 항진균 활성을 유지하였으므로 대단히 안정한 물질인 것으로 판단되었다. 따라서 Bacillus subtilis AF4는 다양한 곰팡이유래 식물질병방제에 적용가능성이 높은 균주임을 알 수 있었다. In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40°C) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum-antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease- stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

콩 종류에 따른 청국장의 발효특성

이나리 ( Na Ri Lee ),이상미 ( Sang Mee Lee ),고태훈 ( Tae Hun Go ),정성윤 ( Seong Yun Jeong ),홍창오 ( Chang Oh Hong ),김근기 ( Keun Ki Kim ),박현철 ( Hyean Cheal Park ),이상몽 ( Sang Mong Lee ),김용균 ( Young Gyun Kim ),손홍주 ( 한국환경과학회 2013 한국환경과학회지 Vol.22 No.6

This study was carried out to investigate changes of protease and amylase activities and nitrogen content in Chungkookjang prepared by Bacillus subtilis S8 and different soybean. Amino-type nitrogen and ammonia-type nitrogen contents increased with an increase in fermentation time and was the highest in black soybean Chungkookjang. The number of viable cells increased up to 24 h of fermentation at all temperatures tested; especially, their levels were the highest at 40℃. Protease activity was the highest in black soybean Chungkookjang. α-amylase activity increased significantly up to 6 h of fermentation at 30℃ and 40℃ and then maintained constantly. It also increased up to 30-36 h of fermentation at 45℃ and then decreased. β -amylase activity was the highest in black soybean Chungkookjang at 35℃and 40℃ and in yellow soybean Chungkookjang at 45℃. Production pattern of reducing sugar was similar to that of β-amylase. Amino-type nitrogen, viable cell number and reducing sugar content and β-amylase activity was the highest in Chungkookjang fermented at 40℃. Considering amino-type and ammonia-type nitrogen contents, Chungkookjang fermentation using yellow soybean was favorable. However, the fermentation using black soybean was favorable, considering protease and amylase activities and reducing sugar content.

유류오염 토양 내 석유계 탄화수소 화합물의 분해에 대한 퇴비의 시용 효과

김성은 ( Sung Un Kim ),김용균 ( Yong Gyun Kim ),이상몽 ( Sang Mong Lee ),박현철 ( Hyean Cheal Park ),김근기 ( Keun Ki Kim ),손홍주 ( Hong Joo Son ),노용동 ( Yong Dong Noh ),홍창오 ( Chang Oh Hong ) 한국환경농학회 2015 한국환경농학회지 Vol.34 No.4

BACKGROUND: Petroleum-contaminated soil from leaking above- and underground storage tanks and spillage during transport of petroleum products is widespread environmental problem in recent years. Application of compost may be the most promising, cost-effective, and eco-friendly technology for soil bioremediation because of its advantages over physical and chemical technology. The objective of this study was to evaluate effect of compost application on degradation of total petroleum hydrocarbon (TPH) in petroleum hydrocarbon-contaminated soil.METHOD AND RESULTS: An arable soil was artificially contaminated by diesel, and compost was applied at the different rate of 0, 10, 30, and 50 Mg/ha. Concentration of TPH in the soil decreased as application rate of compost increased. Degradation efficiency was highest at compost 30 Mg/ha; however, it slightly decreased with compost 50 Mg/ha. Kinetic modeling was performed to estimate the rates of chemical reaction. The correlation coefficient (R2) values for the linear plots using the second-order model were higher than those using the first-oder model. Compost 30 and 50 Mg/ha had the fastest TPH degradation rate in the second-order model. Change of microbial population in soil with compost application was similar to that of TPH.Microbial population in the soil increased as application rate of compost increased. Increasing microbial population in the contaminated soil corresponded to decreased in TPH concentration.CONCLUSION: Conclusively, compost application for soil bioremediation could be an effective response to petroleum hydrocarbon-contaminated soil. The increase in microbial population with compost suggested that compost application at an optimum rate might enhance degradation of TPH in soil.

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.

Pseudomonas sp. LG2의 Ferulic acid esterase 및 Xylanase 유도와 부분적 특성

김용균,이상몽,박현철,김근기,손홍주,Kim, Yong-Gyun,Lee, Sang-Mong,Park, Hyun-Chul,Kim, Keun-Ki,Son, Hong-Joo 한국생명과학회 2007 생명과학회지 Vol.17 No.4

리그닌 분해 세균인 Pseudomonas sp. LG2는 lignocellulose 기질을 분해하여 APPL 화합물을 생성하는 균주이다. 이 균주를 BSG(brewer's spent grain)가 함유된 배지에서 배양한 배양액에서 APPL 화합물을 확인하였다. 세포외 조효소들의 유도에 관한 여러 가지 탄소원의 영향을 조사한 결과 glucose 배지에서는 xylanase의 효소활성만 확인 되었고 xylose, arabinose에서 배양한 조효소에서는 FAE 및 xylanase의 효소활성이 없었다. Oat spelt xylan, HBSG I(hydrolyzed brewer's spent grain I), HBSG II(hydrolyzed brewer's spent grain II) 및 AFBSG(autoclaved fraction from brewer's spent grain)를 탄소원으로 배양한 조효소에서는 FAE 및 xylanase의 효소활성이 확인됐다. Pseudomonas sp. LG2를 oat spelt xylan, HBSG I, HBSG II 및 AFBSG를 탄소원으로 사용하여 14일 동안 배양하면서 배양기간에 따른 세포외 효소들의 FAE와 xylanase 활성을 조사하였다. Xylanase의 최고 활성은 xylan을 탄소원으로 6일간 배양 했을때 5.3 U/mg으로 가장 높았으며, FAE의 최고 활성은 AFBSG를 탄소원으로 배양했을 때 배양 8일째 15.4 mU/mg으로 가장 높았다. Oat spelt xylan, HBSG I, HBSG II 및 AFBSG를 탄소원으로 사용하여 배양한 배지에 분리된 ferulic acid가 확인되었다. 세포외 효소의 FAE 활성은 methyl ferulic acid, methyl caffeic acid, methyl p-coumaric acid에 대해 esterase의 활성을 보였으나, methyl sinapinic acid, methyl vanillic acid 및 methyl gallic acid에 대해서는 esterase의 활성이 없었다. Lignin degrading bacterium Pseudomonas sp. LG2 was able to degrade lignin substrate to a lot of APPL compound. APPL compound was detected in culture supernatants from Pseudomonas sp. LG2 grown with BSC(brewer's spent grain). FAE(ferulic acid esterase) and xylanase are induced from Pseudomonas sp. LG2 in the presence of carbon sources such as oat spelt xylan, HBSG I, II(hydrolyzed brewer's spent grain I, II) and AFBSG(autoclaved fraction from brewer's spent grain). However, xylanase and FAE are not induced by growth of Pseudomonas sp. LG2 on xylose and arabinose. Pseudomonas sp. LG2 is grown on medium containing oat spelt xylan, HBSG I, II and AFBSG and the induction of FAE and xylanase activities of extracellular proteins determined during 14 days. Maximum level of xylanase activity(5.3 U/mg) found at 6 days in culture contained oat spelt xylan as carbon source, whereas maximum level of FAE activity(15.4 mU/mg) was found at 8 days in culture contained AFBSG as carbon source. Most ferulic acid was released in culture supernatants when Pseudomonas sp. LG2 grown on oat spelt xylan, HBSG I, II and AFBSG. FAE of extracellular enzymes was also specific activity on methyl ferulic acid, methyl caffeic acid and methyl p-coumaric acid respectively, but not methyl sinapinic acid, methyl vanillic acid and methyl gallic acid.

Pseudomonas sp. RRj 228이 분비하는 항균물질의 동정과 고추탄저균 C. acutatum에 대한 항균활성

전상윤(Sang-Yoon Jeon),김용균(Yong Gyun Kim),손홍주(Hong Joo Son),박현철(Hyean Cheal Park),이상몽(Sang Mong Lee),김선태(Sun-Tae Kim),박기도(Ki Do Park),강위금(Ui Gum Kang),김근기(Keun Ki Kim) 한국생명과학회 2010 생명과학회지 Vol.20 No.8

식물 근권미생물은 식물의 뿌리 표면과 내부에 정착하면서 식물생장 촉진물질을 분비하여 식물생육에 직접적인 영향을 주거나, 항균물질을 분비하여 병원균으로부터 식물 근권환경을 보호하여 식물의 생육을 촉진하게 된다. 이러한 활성을 갖고 있는 미생물을 분리하여 동정하며, 항균물질을 분리, 동정하여 지속농업의 발전과 농업환경의 보전을 위하여 연구를 실시하였다. 논토양으로부터 식물생육촉진활성과 항균활성을 갖는 RRj 228균주를 분리하여, 생리ㆍ생화학적방법과 유전학적 방법으로 동정한 결과, Pseudomonas sp.로 동정되었다. RRj 228균주는 B. cinerea, P. ultimum, P. capsici와 R. solani에도 높은 항균활성을 보였으며, 특히 고추탄저병원균인 C. acutatum에 강력한 항균활성을 나타냈다. RRj 228균의 배양여액을 C. acutatum, R. solani 및 P. ultimum에 대한 ED50값을 측정한 결과 0.14, 0.16, 0.29 ㎎/ml로 나타났다. 항균활성물질은 RRj 228 배양여액으로부터 각종 크로마토그라피법으로 순수분리하고, NMR과 GC/MS 등의 기기분석을 실시하여 구조를 Phenazine-1-carboxylic acid (분자량 224)로 동정하였다. Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the ED50 value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 ㎎/ml, 0.16 ㎎/ml and 0.29 ㎎/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The ¹H and <SUP>13</SUP>C assignment of the antagonistic substance was achieved from two-dimensional ¹H-¹H COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid (C13H?N₂O₂, M.W.=224).

Responses of Plasma IGF-1, IGFBPs and Hepatic GH Receptor to Growth Hormone Releasing Peptides (GHRP)-2 Administration and Energy Level in Wethers

Hong Gu Lee(이홍구),Young Cheng Jin(김영성),Hisashi Hidari,Yun Jaie Choi(최윤재),Seon Ku Kim(김선구),Teak Soon Shin(신택순),Byung Uuk Cho(조병욱),Yong Gyun Kim(김용균),Keun Ki Kim(김근기),Hong Joo Son(손홍주),Sang Mong Lee(이상몽) 한국생명과학회 2008 생명과학회지 Vol.18 No.7

본 연구는 정상으로 단백질을 급여한 거세면양에 있어서 에너지 첨가가 GHRP-2투여에 대한 혈장 IGF-1 및 IGFBPs에 대한 반응과, 고에너지 급여에 따른 GHRP-2투여가 hepatic GH 수용체에 미치는 영향을 검증하기 위하여 실시하였다. 시험 결과 HENP (CP 0.34 ㎏, TDN 1.83 ㎏/day DM intake)처리기간 동안 혈장 IGF-1 과 39-42kDa IGFBP-3수준은 LENP (CP 0.32 ㎏, TDN 0.87 ㎏/day DM intake)기간에 비하여 높게 나타났으나 (P<0.05), 혈장 34 kDa IGFBP-2와 24 kDa IGFBP-4는 영양처리에 의해 영향을 받지 않았다. 각 영양처리 기간동안 GHRP-2 (12.5 μg/㎏ body weight/day)투여는 혈장 GH 반응이 촉진되었으며(P<0.05), 혈장 GH 평균 함량과 AUC증가에 있어서는 LENP처리 기간에 비하여 HENP처리기간에서 유의적으로 높게 나타났다(P<0.01). 특히 HENP에서 7일간 GHRP-2투여에 의한 일중 혈장 IGF-1 변화양상을 조사한 결과 투여 2, 6 및 7일에서 뚜렷한 증가양상을 보였다(P<0.05). 이에 반하여 LENP에서는 오직 투여 3일째에서 Saline구에 비하여 유의적인 증가를 확인하였다(P<0.05). IGFBPs의 ligand blotting 결과 HENP구에서 혈장 39-43 kDa IGFBP-3의 수준의 증가가 투여 6일과 7일에서 관찰되었으나 혈장 IGFBP-2수준은 두 영양처리시기에서 유의적인 차이를 관찰하지 못했다. 아울러 HENP구에 있어서 간세포막에 <SUP>125</SUP>I-oGH의 결합력을 측정한 결과 GHRP-2투여에 의한 영향은 관찰되지 않았다. 이와 같은 결과는 거세면양에 있어서 단백질과 에너지 사이의 영양적 균형은 내인성 GH/IGF-1 axis는 물론 혈장 IGFBP-3수준의 변화에 영향을 미치고 있음을 시사한다. The purpose of this study was to determine the effect of energy supplement on responses of plasma insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBPs) to growth hormone-releasing peptide-2 (GHRP-2) administration in normal protein-fed wethers, and to observe the effect of GHRP-2 treatment on hepatic growth hormone (GH) receptor in well-fed wethers. Plasma IGF-1 and 39-42 kDa IGFBP-3 during the HENP (CP, crude protein 0.34 and TDN, total digestible nutrients 1.83 ㎏/day DM, dry matter intake) treatment period were higher than in the LENP (CP 0.32 ㎏ and TDN 0.87 ㎏/day DM intake) period (P<0.05). The response of GH was stimulated by GHRP-2 (12.5 ㎍/㎏ body weight/day) administration during both of the feed treatment periods (P<0.05). The area under curve (AUC) increment and average concentration of GH (0-180 min) with GHRP-2 administration was higher during HENP treatment than LENP treatment (P<0.01). During the HENP treatment period from day 1 to day 7 of twice daily GHRP-2 treatment, the plasma IGF-1 increment was increased on days 2, 6 and 7 of GHRP-2 administration (P<0.05). On the basis of ligand blotting, the proportions of plasma 39-43 kDa IGFBP-3 during the HENP treatment period only showed a significant difference on days 6 and 7 with GHRP-2 administration. No significant difference in the specific binding of 125I-labeled oGH to hepatic membranes was detected between the saline and GHRP-2 treatments of the HENP-fed wethers. These results suggest that the nutritional balance between energy and protein may affect the endogenous GH / IGF-1 axis as well as plasma IGFBP-3 levels.