A spectrophotometric assay for the detection of fungal peroxygenases.
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.
Department of Bio- and Environmental Sciences, International Graduate School of Zittau, Markt 23, 02763 Zittau, Germany.
Marzena
M
Poraj-Kobielska
Matthias
M
Kinne
René
R
Ullrich
Katrin
K
Scheibner
Martin
M
Hofrichter
D002396:Catechols / Q000378:metabolism; D004149:Dioxoles / Q000378:metabolism; D005658:Fungi / Q000201:enzymology*; D057166:High-Throughput Screening Assays / Q000379:methods; D006863:Hydrogen-Ion Concentration; D006899:Mixed Function Oxygenases / Q000032:analysis* / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods*
D002396:Catechols; D004149:Dioxoles; D006899:Mixed Function Oxygenases; C022333:peroxygenase; C040539:1,3-benzodioxole; C001833:4-nitrocatechol