Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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An FMN-containing NADH-quinone reductase from streptomyces sp

An FMN-Containing NADH-Quinone Reductase from Streptomyces sp.

  • Youn, Hong-Duk (Research Center for Molecular Microbiology, Seoul National University) ;
  • Lee, Jin-Won (Research Center for Molecular Microbiology, Seoul National University) ;
  • Youn, Hwan (Research Center for Molecular Microbiology, Seoul National University) ;
  • Lee, Jeong-Kug (Seogang University) ;
  • Hah, Yung-Chil (Research Center for Molecular Microbiology, Seoul National University) ;
  • Kang, Sa-Ouk (Research Center for Molecular Microbiology, Seoul National University)
  • Published : 1996.06.01
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Abstract

NADH-quinone reductase was purified 22-fold from the cytosolic fraction of Streptomyces sp. Imsnu-1 to apparent hemogenity, with an overall yield of 9%, by the purification procedure consisting of ammonium, sulfate precipitation and DEAE Sephacryl S-200 and DEAE 5 PW chromatographies. Thes molecular mass of the enzyme determined by gel filtration chromatography was found to be 110 kDa. SDS-PAGE revealed that the enzyme consists of two sugunits with a molecular mass of 54 kDa. The enzyme contained 1 mol of FMN per subunit as a cofactor. The $A_{272}$ A$_{457}$ ratio was 6.14 and the molar extinction coefficients were calculated to be 20, 800 and 25, 400M$^{-1}$ $cm^{-1}$ / AT 349 AND 457 nm, respectively. The N-terminal sequence of the enzyme contained the highly conserved fingerprint of ADP-binding domain. The enzyme used NADH as an electron donor and various quinones as electron acceptors. Cytochrome c was practically inactive. Air-stable flavin semiquinone was produced by the addition of NADH to the enzyme. Also, naphthosemiquinone was detected in the reaction mixture containing the enzyme.

Keywords

References

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