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. 1995 Aug;61(8):3076–3081. doi: 10.1128/aem.61.8.3076-3081.1995

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

B J Brock, S Rieble, M H Gold
PMCID: PMC1388558  PMID: 16535104

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.

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Selected References

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