Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1

Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석

  • Lim Hyun Sook (Graduate School of Education, Chosun University) ;
  • Goo Jae Whan (Department of Environmental engineering, Chosun University) ;
  • Kim Lee Hyun (Graduate School of Education, Chosun University) ;
  • Kim Si Wouk (Department of Environmental engineering, Chosun University) ;
  • Cho Eun Hee (Department of Biology Education, Chosun University)
  • Published : 2005.03.01
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Abstract

Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.

Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP)은 단일 탄소원, 질소원 그리고 에너지원으로 난분해성 화합물인 트리메틸아민을 이용할 수 있다. M. aminosulfidovorans SK1는 진핵세포의 flavin-containing monooxygenase와 유사한 유전자(bFMO)를 지니고 있으며 대장균에서 발현된 재조합 단백질은 강력한 트리메틸아민 산화활성을 보인다. 본 연구에서는 bEMO의 기능과 조절 메커니즘을 연구하기 위하여 bfmo의 상단부 및 하단부 유전자의 염기서열을 결정하였다. bfmo 상단부의 세 개의 열린해독틀은 잘 보존된 nitrate/nitrite response regulators와 methyl accepting protein 유사단백질을 암호화하였다. 하단부의 두 개의 작은 열린해독틀은 기능은 알려져 있지 않지만 진정세균계에서 잘 보존된 단백질의 일종으로 나타났다. 역전사효소 중합효소증폭반응을 통하여 여섯 개의 유전자는 세 개의 독립된 오페론으로 구성되어 있음을 확인하였다. bfmo의 상단부에 위치하는 세 개의 조절유전자는 두 개의 프로모터에서 전사되었다. 그리고 이와 독립적으로 bfmo와 두 개의 하단부 유전자가 하나의 전사단위를 이루고 있다.

Keywords

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