한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제31권1호
  • /
  • Pages.75-82
  • /
  • 2003
  • /
  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

폭약 Hexahydro-1,3,5-trinitro-1,3,5-triazine(RDX)에 노출된 분해세균 Pseudomonas sp. HK-6의 세포반응과 형태변화

Cellular Responses and Morphological Changes of RDX-degrading Bacterium, Pseudomonas sp. HK-6 Exposed by Explosive Hexahydro-1,3,5-triaitro-1,3,5-triazine (RDX).

  • 장효원 (순천향대학교 생명과학부) ;
  • 강형일 (순천대학교 환경교육과) ;
  • 김치경 (충북대학교 미생물학과) ;
  • 오계헌 (순천향대학교 생명과학부)
  • 발행 : 2003.03.01
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초록

환경 오염원으로서 폭약 hexahydro-1,3,5-trinitro-1,3,5-triazine(RDX)에 대한 RDX 분해세균 Pseudomonas sp. HK-6의 세포반응과 형태변화에 대하여 조사하였다. 아치사조건의 RDX농도와 노출시간에 따른 균주 HK-6의 생존율을 분석한 결과, 이 세균의 생존율은 스트레스 충격 단백질의 생성과 비례하였다. 총세포 지방산 조성분석에서 균주 HK-6는 trypticase soy agar(TSA)에서 자랄 때 보다 RDX배지에서 자랄 때 여러 가지 종류의 지방산이 생성되거나 사라지는 것이 밝혀졌다. Anti-DnaK와 anti-GroEL을 이용하여 SDS-PAGE와 Western blot을 통한 분석으로 균주 HK-6는 70 kDa DanK와 60 kDa GroEL을 포함하는 몇가지 스트레스 충격단백질을 생성하는 것으로 밝혀졌다. RDX에 노출된 HK-6배양에서 수용성 단백질 분획에 대하여 2-D PAGE를 실시하였으며, pH 3에서 pH 10 범위에서 약 300 spots가 silver로 염색된 gel상에서 관찰되었다. 그 결과, RDX에 대한 반응으로 10여개의 spots가 현저히 유도 발현되었다. RDX(0.135mM, 12시간)에 노출된 세포는 구멍이 나타나고 표면의 불규칙적인 형태 변화가 일어나 죽게되는 것이 주사 전자현미경을 통하여 관찰되었다.

The cellular responses of RDX-degrading bacterium, Pseudomonas sp. HK-6 to explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) were examined. Strain HK-6 grown at different RDX concentrations was found to demonstrate the survival rate in proportional to the rate of the stress shock proteins produced in this bacterium. Analysis of total cellular fatty acid acids showed that lipids 10:0 iso and 14:1 $\omega$5c/$\omega$5t increased approx three times in strain HK-6 grown on RDX media than TSA media. SDS-PAGE and Western blot using anti-DnaK and GroEL revealed that several stress shock proteins including 70 kDa DnaK and 60 kDa CroEL were newly synthesized in strain HK-6 exposed to different RDX concentrations in exponentially growing cultures. 2-D PAGE of soluble protein fractions from the culture of HK-6 exposed to RDX demonstrated that approximately 300 spots were observed on the silver stained gel ranging from pH 3 to pH 10. As a result, 10 spots were significantly induced and expressed in response to RDX. Scanning electron microscopy fur the cells treated with 0.135 mM RDX for 12 hrs showed the presence of perforations and irregular rod shapes with wrinkled surfaces.

키워드

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