Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Ferric iron reductase activity of LuxG from Photobacterium leiognathi

Photobacterium leiognathi LuxG 단백질의 철(III) 이온 환원 효소 활성도

  • Lee, Eui Ho (Department of Biochemistry, Chungnam National University) ;
  • Nam, Ki Seok (Department of Biochemistry, Chungnam National University) ;
  • Lee, Seon Kwang (Department of Biochemistry, Chungnam National University) ;
  • Oh, Eugeney (Department of Biochemistry, Chungnam National University) ;
  • Lee, Chan Yong (Department of Biochemistry, Chungnam National University)
  • Received : 2016.09.29
  • Accepted : 2016.12.15
  • Published : 2016.12.31
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Abstract

In order to identify the biochemical characteristics of LuxG, the luxG gene from bioluminescence bacteria of Photobacterium leiognathi ATCC 25521 was isolated by PCR-Amplification and inserted into pQE30 vector containing the T5 promoter and 6X His-tag system. The resulting recombinant plasmid was transformed into Escherichia coli to over-express the luxG gene and purify the gene product. The purified LuxG protein demonstrated ferric iron reductase activity and the kinetic parameters of $K_m$ and $V_{max}$ for FMN as well as the NADPH substrates of ferric iron reductase were determined, respectively.

본 연구에서는 발광 세균에 존재하는 LuxG 단백질의 효소학적 성질을 알아내기 위하여 Photobacterium leiognathi ATCC 25521의 luxG 유전자를 중합효소연쇄반응으로 증폭시켜 T5 프로모터와 6X His-tag 시스템을 지닌 pQE 벡터에 삽입시킨 재조합플라스미드를 제조하여 대장균에 형질전환 후 과발현시켜 단백질을 분리, 정제 하였다. 정제된 단백질의 효소학적 실험 결과, 이 단백질은 FMN과 NADPH 기질에 대한 ferric iron reductase의 기능을 갖고 있음을 확인하였으며 이들 기질에 대한 효소 활성도 상수 $K_m$ 및 $V_{max}$ 값을 결정하였다.

Keywords

References

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