Analytical Biochemistry 2012-02-01

A spectrophotometric assay for the detection of fungal peroxygenases.

Marzena Poraj-Kobielska, Matthias Kinne, René Ullrich, Katrin Scheibner, Martin Hofrichter

Index: Anal. Biochem. 421(1) , 327-9, (2012)

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Abstract

Rapid and simple spectrophotometric methods are required for the unambiguous detection of recently discovered fungal peroxygenases in vivo and in vitro. This paper describes a peroxygenase-specific assay using 5-nitro-1,3-benzodioxole as substrate. The product, 4-nitrocatechol, produces a yellow color at pH 7, which can be followed over time at 425 nm (ε(425)=9,700 M(-1) cm(-1)), and a red color when adjusted to pH >12, which can be measured in form of an end-point determination at 514 nm (ε(514)=11,400 M(-1) cm(-1)). The assay is suitable for detecting peroxygenase activities in complex growth media and environmental samples as well as for high-throughput screenings.Copyright © 2011 Elsevier Inc. All rights reserved.