Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Purification and Characterization of NADPH-Dependent Cr(VI) Reductase from Escherichia coli ATCC 33456

  • Bae, Woo-Chul (Division of Bioscience and Bioinformatics, Myongji University) ;
  • Lee, Han-Ki (Division of Bioscience and Bioinformatics, Myongji University) ;
  • Choe, Young-Chool (Division of Bioscience and Bioinformatics, Myongji University) ;
  • Jahng, Deok-Jin (Department of Environmental Engineering and Biotechnology, Myongji University) ;
  • Lee, Sang-Hee (Division of Bioscience and Bioinformatics, Myongji University) ;
  • Kim, Sang-Jin (Microbiology Laboratory, Korea Ocean Research and Development Institute) ;
  • Lee, Jung-Hyun (Microbiology Laboratory, Korea Ocean Research and Development Institute) ;
  • Jeong, Byeong-Chul (Division of Bioscience and Bioinformatics, Myongji University)
  • Published : 2005.02.28
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Abstract

A soluble Cr(VI) reductase was purified from the cytoplasm of Escherichia coli ATCC 33456. The molecular mass was estimated to be 84 and 42 kDa by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, indicating a dimeric structure. The pI was 4.66, and optimal enzyme activity was obtained at pH 6.5 and $37^{\circ}C$. The most stable condition existed at pH 7.0. The purified enzyme used both NADPH and NADH as electron donors for Cr(VI) reduction, while NADPH was the better, conferring 61% higher activity than NADH. The $K_m$ values for NADPH and NADH were determined to be 47.5 and 17.2 umol, and the $V_max$ values 322.2 and 130.7 umol Cr(VI) $min^{-1}mg^{-1}$ protein, respectively. The activity was strongly inhibited by N-ethylmalemide, $Ag^{2+},\;Cd^{2+},\;Hg^{2+}$, and $Zn^{2+}$. The antibody against the enzyme showed no immunological cross reaction with those of other Cr(VI) reducing strains.

Keywords

References

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