Detection of m-toluate in Soils using Bioluminescence Producing Recombinant Bacteria

유전자 재조합 발광균주를 이용한 토양 오염원 m-toluate 탐지

  • Kong, In-Chul (School of Civil, Urban and Environmental Engineering, Yeungnam University) ;
  • Kim, Myung-Hee (School of Civil, Urban and Environmental Engineering, Yeungnam University) ;
  • Jung, Yun-Ho (School of Civil, Urban and Environmental Engineering, Yeungnam University) ;
  • Ko, Kyung-Seok (Geo-Environmental Division, Korea Institute of Geoscience and Mineral Resources) ;
  • Kim, Jae-Gon (Geo-Environmental Division, Korea Institute of Geoscience and Mineral Resources) ;
  • Shin, Sung-Chun (Geo-Environmental Division, Korea Institute of Geoscience and Mineral Resources)
  • 공인철 (영남대학교 건설환경공학부 환경공학) ;
  • 김명희 (영남대학교 건설환경공학부 환경공학) ;
  • 정윤호 (영남대학교 건설환경공학부 환경공학) ;
  • 고경석 (한국지질자원연구원) ;
  • 김재곤 (한국지질자원연구원) ;
  • 신성천 (한국지질자원연구원)
  • Published : 2005.05.31

Abstract

This research focuses on the development and application of a method for the detection of m-toluate in soils using a genetically engineered bioluminescent bacteria, Pseudomonas putida mt-2 KG1206. KG1206 produces light by direct (m-toluate and benzoate) and indirect (toluene analogs) inducers. For detection of m-toluate in soil system, 9.9 mL strain was amended with 0.1 mL soil ethanol extractant. A high correlation ($r^2>0.97$) was observed between bioluminescence and m-toluate concentration. The unknown concentrations of m-toluate in soil samples were pre-determined using a method developed based on bioluminescence activity of strain with extracted inducers. Values between by LC analysis and bioluminescence activity show moderate statistical results. These results demonstrate the feasibility of recombinant bioluminescent microorganism, engineered to generate a quantifiable bioluminescence signal in response to specific pollutants, may serve as combined sensing and reporting tools in environmental monitoring.

본 연구에서는 유전자 재조합 발광균주, Pseudomonas putida mt-2 KG1206을 이용하여 토양에 오염된 m-toluate 탐지 방법 및 적용 가능성에 대해 조사하였다. KG1206은 톨루엔 계열 화합물의 중요 중간 분해물질인 m-toluate 및 benzoate가 직접 생물 발광 유도제로 작용하며, 또한 톨루엔 계열 화합물들이 간접 유도제로서 발광 활성을 나타내었다. 토양에 오염된 유도제 오염원 검출을 위해 발광 균주 9.9 mL에 에탄올 추출물 0.1 mL을 첨가하여 조사하였다. 생물발광에 근거하여 작성된 m-toluate 검량선은 대략 $R^2>0.97$ 이상의 상관관계가 관찰되었다. 토양에 임의 오염된 m-toluate(직접 발광유도제)는 정립한 방법에 따라 발광활성에 근거하여 추측하였고, 기기분석치와 통계적으로 유의한 것으로 조사되었다. 본 연구 결과를 통해서 특정 화합물에 대해 발광을 생산하는 유전자 재조합 균주가 특정 오염원에 오염된 지역의 관리를 위한 수단으로 사용할 수 있는 가능성을 확인할 수 있었다.

Keywords

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