Open AccessPhenolic substrates for fluorometric detection of laccase activity
Author(s) -
Lonergan Greg,
Mew Elizabeth,
Schliephake Kirsten,
Baker Warren L
Publication year - 1997
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1997.tb12614.x
Subject(s) - laccase , chemistry , homovanillic acid , substrate (aquarium) , chromatography , fluorescence , enzyme , biochemistry , biology , ecology , physics , receptor , quantum mechanics , serotonin
A fluorometric procedure has been developed for detection and estimation of laccase activity in fungal broth cultures. Laccase solution was pretreated with catalase for 1 h at 37°C and pH 5. Homovanillic acid was then added and the reaction mixture incubated for a further hour at 37°C. The fluorescence was then developed by addition of 0.1 M glycine buffer at pH 10. Laccase preparations from Pyricularia oryzae, Coriolus hirsutus and Pycnoporus cinnabarinus catalysed formation of a fluorescent product of HVA but the optimum pH values of enzyme activities varied. The culture fluids of several other fungi also catalysed development of fluorescence in solutions containing HVA. p ‐Hydroxyphenylacetic acid was a poor substrate for all laccases in vivo except that produced by Perennipora tephropora .