Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Partial Cloning of Genes for Lignin Degrading Enzymes in Trametes versicolor

구름버섯에서 리그닌 분해효소 유전자들의 클로닝

  • 김용호 (강원대학교 생명과학부 미생물학과) ;
  • 정수진 (강원대학교 생명과학부 미생물학과) ;
  • 김선경 (강원대학교 생명과학부 미생물학과) ;
  • 송홍규 (강원대학교 생명과학부 미생물학과) ;
  • 최형태 (강원대학교 생명과학부 미생물학과)
  • Published : 2003.09.01
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Abstract

Laccase, lignin- and manganese peroxidase are implicated in the lignin degradation. The nucleotide sequences of four copper-binding domains in fungal laccases, and heme-binding domains of lignin- and manganese peroxidases are well conserved, and therefore these short fragments can be used for the PCR for the gene amplification. We synthesized several PCR primers according to their sequences, and run PCR to amplifiy the lignin degrading genes of Trametes versicolor isolated in Korea. PCR products were cloned with pGEM-T vector in order to determine their nucleotide sequences. A laccase fragment (1.3 kb) showed 65-97％ homologies, lignin peroxidase fragment (185 bp) showed 80-95％ homologies, and manganese peroxidase fragment (443 bp) showed 61-83％ homologies when compared with other white-rot fungal enzymes.

리그닌 분해에 관련된 효소 중 laccase는 구리 결합부위가, lignin peroxidase와 manganese peroxidase는 헴(heme) 결합부위의 아미노산 서열이 잘 보존된 단백질들이다. 국내에서 분리한 구름버섯(Trametes versicolor)을 대상으로 관련 유전자들의 보존된 지역의 염기서열을 근거로 중합연쇄반응(PCR)에 필요한 primer를 제작하였고, 이를 사용하여 해당 유전자의 조각을 확보하였다. 이를 pGEM-T vector에 cloning하고 그 염기서열을 분석한 결과 약 1.3 kb의 laccase조각은 다른 백색부후균의 laccase와 65～97％의 상동성을 보였다. 또한 185 bp의 lignin peroxidase 조각과 443 bp의 manganese peroxidase조각을 타 백색부후균의 효소들과 비교한 결과 각각 80～95％ 및 61～83％의 상동성을 보였다.

Keywords

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