Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Purification and Characterization of an Intracellular NADH: Quinone Reductase from Trametes versicolor

  • Lee, Sang-Soo (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University) ;
  • Moon, Dong-Soo (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University) ;
  • Choi, Hyoung-T. (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University) ;
  • Song, Hong-Gyu (Division of Life Sciences, and Research Institute of Life Sciences, Kangwon National University)
  • Published : 2007.08.30
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Abstract

Intracellular NADH:quinone reductase involved in degradation of aromatic compounds including lignin was purified and characterized from white rot fungus Trametes versicolor. The activity of quinone reductase was maximal after 3 days of incubation in fungal culture, and the enzyme was purified to homogeneity using ion-exchange, hydrophobic interaction, and gel filtration chromatographies. The purified enzyme has a molecular mass of 41kDa as determined by SDS-PAGE, and exhibits a broad temperature optimum between $20-40^{\circ}C$, with a pH optimum of 6.0. The enzyme preferred FAD as a cofactor and NADH rather than NADPH as an electron donor. Among quinone compounds tested as substrate, menadione showed the highest enzyme activity followed by 1,4-benzoquinone. The enzyme activity was inhibited by $CuSO_4,\;HgCl_2,\;MgSO_4,\;MnSO_4,\;AgNO_3$, dicumarol, KCN, $NaN_3$, and EDTA. Its $K_m\;and\;V_{max}$ with NADH as an electron donor were $23{\mu}M\;and\;101mM/mg$ per min, respectively, and showed a high substrate affinity. Purified quinone reductase could reduce 1,4-benzoquinone to hydroquinone, and induction of this enzyme was higher by 1,4-benzoquinone than those of other quinone compounds.

Keywords

References

  1. Brock, B., S. Rieble, and M. Gold. 1995. Purification and characterization of a 1,4-benzoquinone reductase from the basidiomycete Phanerochaete chrysosporium. Appl. Environ. Microbiol. 61, 3076-3081
  2. Cheong, S., S. Yeo, H.-G. Song, and H.-T. Choi. 2006. Determination of laccase gene expression during degradation of 2,4,6-trinitrotoluene and its catabolic intermediates in Trametes versicolor. Microbiol. Res. 161, 316-320 https://doi.org/10.1016/j.micres.2005.12.001
  3. Cohen, R., M. Suzuki, and K. Hammel. 2004. Differential stress-induced regulation of two quinone reductases in the brown rot basidiomycete Gloeophyllum trabeum. Appl. Environ. Microbiol. 70, 324-331 https://doi.org/10.1128/AEM.70.1.324-331.2004
  4. Conesa, A., P. Punt, and C. van den Hondel. 2002. Fungal peroxidases: molecular aspects and applications. J. Biotechnol. 93, 143-158 https://doi.org/10.1016/S0168-1656(01)00394-7
  5. Constam, D., A. Muheim, W. Zimmermann, and A. Fiechter. 1991. Purification and partial characterization of an intracellular NADH:quinone oxidoreductase from Phanerochaete chrysosporium. J. Gen. Microbiol. 137, 2209-2214 https://doi.org/10.1099/00221287-137-9-2209
  6. Han, M.-J., H.-T. Choi, and H.-G. Song. 2004. Degradation of phenanthrene by Trametes versicolor and its laccase. J. Microbiol. 42, 94-98
  7. Han, M.-J., H.-T. Choi, and H.-G. Song. 2005. Purification and characterization of laccase from the white rot fungus Trametes versicolor. J. Microbiol. 43, 555-560
  8. Haynes, C., R. Koder, A. Miller, and D. Roders. 2002. Structure of nitroreductase in three states. J. Biol. Chem. 277, 11513-11520 https://doi.org/10.1074/jbc.M111334200
  9. Henriksson, G., G. Johansson, and G. Pettersson. 2000. A critical review of cellobiose dehydrogenases. J. Biotechnol. 78, 93-113 https://doi.org/10.1016/S0168-1656(00)00206-6
  10. Ichinose, H., H. Wariishi, and H. Tanaka. 2002. Identification and heterologous expression of the cytochrome P450 oxidoreductase from the white-rot basidiomycete Coriolus versicolor. Appl. Microbiol. Biotechnol. 59, 658-664 https://doi.org/10.1007/s00253-002-1083-8
  11. Jensen, K., Jr., Z. Ryan, A. Wymelenberg, D. Cullen, and K. Hammel. 2002. An NADH:quinone oxidoreductase active during biodegradation by the brown-rot basidiomycete Gloeophyllum trabeum. Appl. Environ. Microbiol. 68, 2699-2703 https://doi.org/10.1128/AEM.68.6.2699-2703.2002
  12. Manchenko, G.P. 1994. Handbook of detection of enzyme on electrophoretic gels. CRC Press p. 84-88
  13. Melo, A., T. Bandeiras, and M. Teixeira. 2004. New insights into type II NAD(P)H:quinone oxidoreductases. Microbiol. Mol. Biol. Rev. 68, 603-616 https://doi.org/10.1128/MMBR.68.4.603-616.2004
  14. Qi, W. and J. Jellison. 2004a. Characterization of a transplasma membrane redox system of the brown rot fungus Gloeophyllum trabeum. Int. Biodet. Biodeg. 53, 37-42 https://doi.org/10.1016/j.ibiod.2003.09.002
  15. Qi, W. and J. Jellison. 2004b. Induction and catalytic properties of an intracellular NADH-dependent 1,4-benzoquinone reductase from the brown-rot basidiomycete Gloeophyllum trabeum. Int. Biodet. Biodeg. 54, 53-60 https://doi.org/10.1016/j.ibiod.2004.02.001
  16. Rasmussen, S., N. Chung, A. Khindaria, T. Grover, and S. Aust. 1995. Reductions catalyzed by a quinone and peroxidases from Phanerochaete chrysosporium. Arch. Biochem. Biophys. 320, 243-249 https://doi.org/10.1016/0003-9861(95)90006-3
  17. Roy, B., T. Dumonceaux, A. Koukoulas, and F. Archibald. 1996. Purification and characterization of cellobiose dehydrogenases from the white rot fungus Trametes versicolor. Appl. Environ. Microbiol. 62, 4417-4427
  18. Shin, K.-S., Y.-H. Kim, and J.-S. Lim. 2005. Purification and characterization of manganese peroxidase of the white-rot fungus Irpex lacteus. J. Microbiol. 43, 503-509
  19. Stahl, J., S. Rasmussen, and S. Aust. 1995. Reduction of quinones and radicals by a plasma membrane redox system of Phanerochaete chrysosporium. Arch. Biochem. Biophys. 322, 221-227 https://doi.org/10.1006/abbi.1995.1455
  20. Vyas, B., J. Volc, and V. Šašek. 1994. Ligninolytic enzymes of selected white rot fungi cultivated on wheat straw. Folia Microbiol. 39, 235-240 https://doi.org/10.1007/BF02814655
  21. Watanabe, M., T. Nishino, K. Takio, T. Sofuni, and T. Nohmi. 1998. Purification and characterization of wild-type and mutant 'classical' nitroreductase of Salmonella typhimurium. J. Biol. Chem. 273, 23922-23928 https://doi.org/10.1074/jbc.273.37.23922
  22. Wesenberg, D., I. Kyriakides, and S. Agathos. 2003. White-rot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol. Adv. 22, 161-187 https://doi.org/10.1016/j.biotechadv.2003.08.011
  23. Xiao, Y., X. Tu, J. Wang, M. Zhang, Q. Cheng, W. Zeng, and Y. Shi. 2003. Purification, molecular characterization and reactivity with aromatic compounds of a laccase from basidiomycete Trametes sp. strain AH28-2. Appl. Microbiol. Biotechnol. 60, 700-707 https://doi.org/10.1007/s00253-002-1169-3