Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of NADH-Dependent Cr(VI) Reductase from Escherichia coli ATCD33456

  • Bae, Woo-Chul (Department of Biological Sciences/BK21 Graduate Program in Environmental and Biological Engineering, MyungJi University) ;
  • Kang, Tae-Gu (Department of Biological Sciences/BK21 Graduate Program in Environmental and Biological Engineering, MyungJi University) ;
  • Jung, Jae-Han (Department of Biological Sciences/BK21 Graduate Program in Environmental and Biological Engineering, MyungJi University) ;
  • Park, Chul-Jae (Department of Biological Sciences/BK21 Graduate Program in Environmental and Biological Engineering, MyungJi University) ;
  • Choi, Sung-Chan (Department of Environmental Science, Hallym University) ;
  • Jeong, Byeong-Chul (Department of Environmental Science, Hallym University)
  • Published : 2000.10.01
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Abstract

A soluble Cr(VI) reductase was purified from the Cr(VI) reducing strain Escherichia coli ATCC33456 by ammonium sulfate fractionation, and chromatographies on Q-Sepharose FF, Cibacron blue 3GA dye affinity, Mono-Q 5/5, and Superdex 200 HR 10/30 columns. The estimated molecular mass of the purified enzyme was 27 kDa on SDS-polyacrylamide gel electrophoresis and 54 kDa on gel filtration, thus indicating a dimeric structure. The isoelectric point of the enzyme was pH 4.85. The optimum reaction pH and storage pH were both 7.0, the optimum reaction temperature was $37^{\circ}C$, and the storage temperature was $4^{\circ}C$. NADH and NADPH both served as electron donors for the reductase, with $V_{max}$ of 68.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 7.6 $\mu$M using HADH, and Vmax of 42.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 14.6 $\muM$ using NADPH. When 1 mM EDTA was added, the Cr(VI) reducing activity increased 4-fold.

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References

  1. Determination of Dissolved Hexavalent Chromium in Drinking Water, Groundwater, and Industrial Wasterwater Effluent by lon chrimatography Arar, E. J.;S. E. Long;J. D. Pfaff
  2. Microbiol v.38 Reduction of hexavalent chromium by Escherichia coli ATCC33456 in batch and continuous cultures Bae, W. C.;T. G. Kang;I. K. Kang;Y. J. Won;B. C. Jeong
  3. Microbiol v.150 Chromate resistance and reduction in Pseudomonas fluorescens strain LB300 Bopp, L. H;H. L. Ehrlich
  4. Biochem v.72 A rapid and sensitive method for the quantitation of protein-dye binding Bradford, M. M
  5. Chromium(Ⅵ)reduction by pseudominas fluorescens LB300 in fixed film bioreactor v.123 Chromium(Ⅵ)reduction by pseudominas fluorescens LB300 in fixed film bioreactor Chira, E. M. N.;Y. T. Wang
  6. technol v.31 Hexavalent chromium reduction by Bacillus sp. in packed bed bioreactor Chirwa, E. M. N.;Y. T. Wang
  7. Microbiologiya v.55 Reduction of hexavalent chromium by collection strains of bacteria Gvozdyak, P. I.;N. F. Mogilevich;A. F. Rylskii;N. M. Grishcheno
  8. Agric. Biol. Chem v.42 Isolation of potassium chromate-tolerant bacterium and chromate uptakes by the bacterium Horitsu, H.;H. Nishida;H. Kato;M. Tomoyeda
  9. Agric. Biol. Chem v.51 Enzymatic reduction of hexavalent chromium by hexavalent chromium tolerant P. ambigua G-1 Horitsu, H.;S. Futo;Y. Miyazawa;S. Ogai;K. Kawai
  10. Appl. Environ. Microbiol v.56 Chromium reduction in Psedomonas putida Ishibashi, Y.;C. Cervantes;S. Silver
  11. Microbiol. Biotechnol v.9 A microbial fuel cell type lactate biosensor using a metal-reducing bacterium, Shewanella putrefaciens Kim, H. J.;M.S. Hyun;i. S. Chang;B. H. Kim
  12. mikrobiologiya v.57 Chromium reducing bacteria in nature and industrial sewage Kvasnikov, E. I.;T. M. Klyushnikova;T. P. Kasatikina;V. V. Stepanyuk;S. L. Kuberskaya
  13. Mikrobiologiya v.54 A new chromium reducing gram variable bacterium with mixed type of flagellation Kvasnikov, E. I.;V. V. Stepanyuk;T. M. Klyushnkova;N. S. Serpokrylov;G. A. Simonova;T. P. Kasatikina;L. P. Panchenko
  14. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature v.227 Laemmli, U. K
  15. Mikrobiologiya v.48 Reduction of crocoite by Pseudomonas chromatophila Lebedeva, E. V.;N. N. Lyalikova
  16. Mutagenic and cytogenetic effectsof chromium compounds Biological and Environmental Aspect of Chromium Levis, A. G.;V. Bianchi
  17. Appl. Environ. Microbiol v.59 Chromate redution by resting cells of Agrobacterium radiobacter EPS-916 Llovera, S.;R. Bonet;M. D. Simon-Pujol;F. Congregado
  18. Appl. Environ. Microbiol v.60 Reduction of chromate by Desulfovibrio vugaris and its c3 cytochrome Lovely, D. R.;E. J. P. Phillips
  19. Microbiol v.35 Redution of hexavalent chromium by P. aeruginisa HP014 Oh, Y. S.;S. C. Choi
  20. Fungal Technologies Vol.2. Bioremediation Congress Redution of hexavalent chromium to trivalent chromium by Phanerochaete chrysosporium Pal, N;In R. Lamar;J. A. Glaser(eds.)
  21. Environ, Microbiol v.66 Purification to homogenety and characterization of a novel Pseudimonas putida shromate reductase Park, C. H.;M. Keyhan;B. Wielinga;S. Fendorf;A. Matin
  22. Microbiol. Biotechnao v.9 Chracteristics of metal biosorption of oxidized Undaria pinnatifida Park, J. Y.;C. Chun;Y. J. Yoo
  23. Industrial Wastewater Treatment Technology Patterson, J.W.
  24. Appl. Environ. Microbiol. v.59 Characterization of enzymatic reduction of hexavalent chromium by Escherichia coli ATCC33456 Shen, H.;Y.T. Wang
  25. J. Environ. Eng. v.120 Biological reduction of chromium by E. coli Shen, H.;Y.T. Wang
  26. J. Microbiol. Biotechnol. v.8 Incidence of multiple heavy metal resistance in a Bacillus species Singh, K.L.;A. Kumar
  27. J. Bacteriol v.174 NAD(P)H-dependent chromium(VI) reductase of P. ambigua G-1: A Cr(VI) intermediate is formed during the reduction of Cr(VI) to Cr(Ⅲ) Suzuki, T.;N. Miyata;H. Horitsu;K. Kawai;K. Takamizawa;Y. Tai;M. Okazaki
  28. Anal. Chem. v.27 Stability of colorimetric reagent for chromium, S-diphenylcarbazide, in various solvents Urone, P.F.
  29. FEMS Microbiol. Lett. v.78 Membrane bound respiratory system of Enterobacter cloacae grown anaerobically with chromate Wang, P.C.;K. Toda;H. Ohtake;I. Kusaka;I. Yabe
  30. Appl. Environ. Microbiol. v.55 Isolation and characterization of an Enterobacter cloacae strain that reduces hexavalent chromium under anaerobic conditions Wang, P.C.;T. Mori;K. Komori;M. Sasatsu;K. Toda;H. Ohtake
  31. J. Bacteriol. v.172 Membrane associated chromate reductase activity from enterobacter cloacae Wang, P.C.;T. Mori;K. Toda;H. Ohtake
  32. Water Res. v.29 Factors affecting hexavalent chromium reduction in pure cultures of bacteria Wang, Y.T.;C. Xiao
  33. J. Ind. Microbiol. v.14 Bacterial reduction of hexavalent chromium Wang, Y.T.;H. Shen