미생물학회지 (Korean Journal of Microbiology)

  • 제41권1호
  • /
  • Pages.13-17
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  • 2005
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Aniline 분해세균 Delftia sp. JK-2에서 분리된 Catechol 2,3-dioxygenase의 N-말단 아미노산 서열 분석

Analysis of N- Terminal Amino Acid Sequence of Catechol 2,3-dioxygenase from Aniline Degrading Delftia sp. JK-2

  • 황선영 (순천향대학교 자연과학대학 생명과학부) ;
  • 강형일 (순천대학교 환경교육과) ;
  • 오계헌 (순천향대학교 자연과학대학 생명과학부)
  • Hwang Seon-Young (Department of Life Science, Soonchunghyang University) ;
  • Kahng Hyung-Yeel (Department of Environmental Education, Sunchon National University) ;
  • Oh Kye-Heon (Department of Life Science, Soonchunghyang University)
  • 발행 : 2005.03.01
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초록

본 연구에서는 이 전 연구에서 단일 탄소원과 질소원 및 에너지원으로 aniline을 이용하는 Delftia sp. JK-2에서 분리, 정제된 바 있는 C2,3O의 N-말단 아미노산과 DNA 서열을 분석하였다. Aniline에서 배양한 Delftia sp. JK-2에서 분리된 약 35kDa의 C2,3O의 N-말단 아미노산 서열을 분석 한 결과 $^1MGVMRIGHASLKVMDMDAAVRHYENV^{26}$로 Pseudomonas sp. AW-2와 Comamonas sp. JS765의 C2,3O와 일치하는 것으로 나타났다. 위에서 확인된 아미노산 서열을 바탕으로 제작된 primer와 JK-2의 total genomic DNA를 기질로 사용하여 PCR을 수행한 결과 약 950 bp의 유전자 증폭산물을 획득하였다. 이 증폭산물 중 정확히 확인된 890 bp의 염기서열을 분석한 결과 Delftia JK-2의 C2,3O유전자 염기서열은 Pseudomonas su. AW-2의 C2,3O와 일치하였으며 Comamonas sp. Js765의 C2,3O와 $97\%$의 높은 상동성을 나타내었다.

The aim of this work was to investigate the N-terminal amino acid sequence of catechol 2,3-dioxygenase isolated from Delftia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O from strain JK-2 was $^1MGVMRIGHASLKVMDMDAAVRHYENV^{26}$, and exhibited high sequence similarity with that of C2,3O from Pseudomonas sp., Comamonas sp. JS765, Comamonas test-osteroni, or Burkholderia sp. RP007. Approximately 950-bp C2,3O was obtained through PCR using the primers derived from N-terminal amino acid sequence. Analysis of the DNA sequence revealed that the deduced 296 amino acid sequences were determined, and it showed $100\%$ identity with C2,3O from Pseudomonas sp. AW-2 and $97\%$ similarity with Comamonas sp. JS765.

키워드

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