Background: Wood rotting white-rot and litter-decomposing basidiomycetes form a huge reservoir of
oxidative enzymes, needed for applications in the pulp and paper and textile industries and for bioremediation.
Objective: The aim was (i) to achieve higher throughput in enzyme screening through miniaturization
and automatization of the activity assays, and (ii) to discover fungi which produce efficient oxidoreductases
for industrial purposes.
Methods: Miniaturized activity assays mostly using dyes as substrate were carried out for lignin peroxidase,
versatile peroxidase, manganese peroxidase and laccase. Methods were validated and 53 species
of basidiomycetes were screened for lignin modifying enzymes when cultivated in liquid mineral, soy,
peptone and solid state oat husk medium.
Results: Manganese peroxidases were the most common enzymes produced by 96% of the species.
They typically had acidic pH optima, although Hyphodontia sp., Pleurotus pulmonarius and Trametes
ochracea produced enzymes highly active at pH 7. Versatile peroxidase was produced by 66% of the
fungi with efficient production from Phlebia radiata, P. pulmonarius and Galerina marginata. Novel
lignin peroxidase producing fungi Cylindrobasidium evolvens and Daedaleopsis septentrionalis were
found among the 26% of the species showing here lignin peroxidase production. Laccase was shown in
92% of the species. Several fungi produced laccase active at pH 7, which is noteworthy because usually
laccases of white-rot fungi are efficient and relevant for many industrial applications.
Conclusion: Automated screening allowed us to monitor many specific enzyme activities and extend the
range of assay conditions from relatively small fungal cultivation sample volumes.