Deciphering the distribution and types of Multicopper oxidases in Basidiomycota fungi

• Phylogeny of Basidiomycota Fungi. • Distribution and Phylogeny of Multicopper Oxidases in Basidiomycota Fungi. • Identification of three novel groups of Laccase-like enzymes. • Classification of Multicopper Oxidases into 8 groups with key structural features. • Reclassification of already reported Multicopper Oxidases. Multicopper oxidases (MCOs) comprise different types of enzymes widely distributed in nature with quite diverse functions. Laccases are the most interesting MCOs from a biotechnological point of view, particularly those secreted by ligninolytic Basidiomycota fungi due to their versatility to oxidize lignin and a variety of aromatic substrates. The term “laccase” has been broadly (but sometimes erroneously) applied due to their low sequence homology and some overlapping activities with other MCO groups. We examined the distribution and phylogenetic relationships of MCOs in Basidiomycota fungi aiming to provide a complete and precise picture of the different MCO types across the division, including fungal orders phylogenetically distant from those typically studied. The phylogenetic tree revealed eight clusters of MCOs, each sharing common sequence/structural features. With this information we classified the MCOs in eight groups and described their distinctive amino acid residues. These eight MCO types are: laccases (LAC), ferroxidases (FOX), laccase-ferroxidases (LAC-FOX), ascorbate oxidases (AO), fungal pigment MCOs, and three new groups of laccase-like enzymes or “atypical laccases” related to but different from laccases sensu stricto, namely novel laccases (NLAC), new MCO (NMCO) and new laccases with potential ferroxidase activity (NLF). Additionally, several MCOs already described in the literature were reclassified into the updated groups.