A simple assay for measuring cellobiose dehydrogenase activity in the presence of laccase.
The commonly used assay for measuring cellobiose dehydrogenase (CDH) activity, based on the reduction of dichlorophenol-indophenol (DCIP), has been adapted to measure this enzyme activity in the presence of laccase, which is often formed concurrently with CDH by a number of fungi. Laccase interferes with the assay by rapidly reoxidizing the reduced form of DCIP and can mask CDH activity completely. It can be conveniently and completely inhibited by 4 mM fluoride in the assay, while CDH activity is only slightly affected by the addition of this inhibitor. The modified assay enables the detection of low CDH activities even in the presence of very high excesses of laccase. It should be useful for screening culture supernatants of wood-degrading fungi for CDH since the assay is rapid and uses inexpensive and nontoxic reagents. Furthermore, it might be used for the detection of other enzyme activities which are assayed by following the reduction of quinones or analogue compounds such as DCIP.
Baminger
U
U
Division of Biochemical Engineering, Institute of Food Technology, Universität für Bodenkultur BOKU (University of Agricultural Sciences Vienna), Wien, Austria.
Nidetzky
B
B
Kulbe
K D
KD
Haltrich
D
D
D001487:Basidiomycota / Q000201:enzymology; D002237:Carbohydrate Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D002733:Chlorophenols / Q000378:metabolism; D005658:Fungi / Q000201:enzymology*; D007215:Indophenol / Q000378:metabolism; D042845:Laccase; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D012969:Sodium Fluoride / Q000494:pharmacology*
D002733:Chlorophenols; D007215:Indophenol; D012969:Sodium Fluoride; D010088:Oxidoreductases; D002237:Carbohydrate Dehydrogenases; C019859:cellobiose-quinone oxidoreductase; D042845:Laccase; C116190:3,5-dichlorophenol