Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library

토양 Metagenome Library로부터 고추역병 저해 클론 탐색

  • Park, Hae-Chul (Department of chemistry, Hanyang university) ;
  • Sung, So-Ra (Department of chemistry, Hanyang university) ;
  • Kim, Dong-Gwan (Research Center for Drugs, Kunkuk University) ;
  • Koo, Bon-Sung (Fermentation & Food Processing Division, Department of Korean Food Research for Globalization, National Academy of Agricultural Science, Rural Development Administration) ;
  • Jeong, Byeong-Moon (Department of Chemistry, Division of Nano Sciences, Ewha Womans University) ;
  • Kim, Jin-Heung (Department of Chemistry, Division of Nano Sciences, Ewha Womans University) ;
  • Yoon, Moon-Young (Department of chemistry, Hanyang university)
  • 박해철 (한양대학교 화학과) ;
  • 성소라 (한양대학교 화학과) ;
  • 김동관 (건국대학교 의약연구센터) ;
  • 구본성 (국립농업과학원 한식세계화연구단 발효이용과) ;
  • 정병문 (이화여자대학교 나노과학부 화학과) ;
  • 김진흥 (이화여자대학교 나노과학부 화학과) ;
  • 윤문영 (한양대학교 화학과)
  • Received : 2009.04.01
  • Accepted : 2009.05.15
  • Published : 2009.06.30
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Abstract

We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

고추 역병을 야기하는 Phytophthora capsici 는 짧은 시간 내에 많은 면적에 피해를 주는 병으로 한번 발생하면 방제가 어려운 병으로 알려져 있다. 이러한 역병 곰팡이의 방제를 위하여 본 연구에서는 P. capsici의 염색체 복제 및 세포 골격 유지 등에 관여하는 단백질인 microtubule의 형성 저해를 유도하는 물질을 탐색하여 궁극적으로 고추역병 방제를 위한 연구를 진행하였다. 먼저 P. capsici alpha 및 beta tubulin을 E. coli BL21(DE3)에서 발현시켜 분리 정제하여 in vitro microtubule 형성을 확인하였다. P. capsici microtubule 형성 저해 metagenome clone 스크리닝을 위하여 경기도 수원의 여기산 토양에서 metagenome을 분리하여 library를 제작하여 Fluorescence Resonance Energy Transfer (FRET) 방법을 이용하여 P. capsici microtubule 형성을 저해하는 화합물을 탐색하였다. In vitro 스크리닝에서 약 384개의 metagenome library에서 2종의 clone을 선택하여 고추작물에 직접 방제하여 역병균의 생장 억제를 확인하였다. 이는 차후 고추역병 방제제 개발에 있어 중요한 후보물질뿐만 아니라 metagenome library를 이용한 새로운 방법의 개발이라 사료 된다. 또한 in vitro 스크리닝에서 얻어진 2종의 metagenome clone의 염기서열을 분석하여 항역병 활성에 관련하는 DNA 서열을 확보하고 이를 응용하여 물질을 생산 할 경우, 현장에서 활용 할 수 있는 효과 큰 친환경 천연고추역병 방제제로서의 개발 가능성을 가진다는 점에서 본 연구결과는 매우 의미 있는 결과라 생각된다.

Keywords

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