Purification and Characterization of a 1,4-Benzoquinone Reductase from the Basidiomycete Phanerochaete chrysosporium.

Applied and Environmental Microbiology
B J BrockM H Gold

Abstract

An intracellular, soluble 1,4-benzoquinone reductase was purified from agitated cultures of Phanerochaete chrysosporium and characterized. The quinone reductase was expressed in cultures grown under both nitrogen-sufficient and nitrogen-limiting (12 and 1.2 mM ammonium tartrate) conditions. The protein was purified to homogeneity by using ammonium sulfate fractionation, hydrophobic interaction, and ion-exchange and blue-agarose affinity chromatographies. The native flavin mononucleotide-containing protein, pI 4.3, has a molecular mass of 44 kDa as determined by gel filtration. The protein has a subunit molecular mass of ;sim22 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The quinone reductase exhibits a broad pH optimum between 5.0 and 6.5 and a temperature optimum of 30(deg)C. The enzyme catalyzes the two-electron reduction of several quinones and other electron acceptors utilizing either NADH or NADPH as an electron donor. The apparent K(infm) for 2-methoxy-1,4-benzoquinone is 2.4 (mu)M, and the apparent k(infcat) is 4.4 x 10(sup5) s(sup-1). Enzyme activity is strongly inhibited by Cibacron blue 3GA and by dicumarol.

References

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Citations

Jul 27, 2001·Applied and Environmental Microbiology·C LeitnerD Haltrich
Jun 1, 2002·Applied and Environmental Microbiology·Kenneth A Jensen JrKenneth E Hammel
Sep 22, 2009·Applied Microbiology and Biotechnology·Clemens K Peterbauer, Jindrich Volc
Mar 29, 2008·FEMS Microbiology Reviews·Petr Baldrian, Vendula Valásková
Apr 21, 2009·Applied and Environmental Microbiology·Víctor Gómez-ToribioFrancisco Guillén
Apr 27, 2010·Chembiochem : a European Journal of Chemical Biology·David B HawkesJames J De Voss
Jun 10, 2015·Proceedings of the National Academy of Sciences of the United States of America·Nicolás PedriniNemat O Keyhani
Jan 30, 1999·Applied and Environmental Microbiology·L AkileswaranM H Gold
Jul 1, 1997·Microbiology·G Vijay Bhasker ReddyMichael H Gold
Nov 4, 2000·Chemosphere·G KöllerK Czihal
May 5, 2006·Journal of Bacteriology·Eric V Patridge, James G Ferry
Oct 24, 2007·Journal of Bacteriology·Susana L A AndradeOliver Einsle
Aug 27, 2021·Journal of Fungi·Mariane DaouCraig B Faulds

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