Abstract
Laccases are part of the family of ligninolytic enzymes and have played essential roles in several biological filamentous fungi processes, including fruiting body formation and lignin degradation. This study aimed to identify and characterize laccase genes in silico of several basidiomycete strains. The applied guaiacol oxidation test allowed the selection of seven out of 11 strains with ligninase activity, which were used for DNA extraction and amplification of the copper-binding region. A single amplicon of approximately 450 bp, was produced by all selected strains and they were further sequenced. Sequence analysis has suggested the presence of a new subdivision of the laccase genes. Clustering analysis confirmed the existence of two groups: cluster A with six strains and singleton B with U8-11 strain. The structural predictions of the U8-11 protein were dissimilar compared to other proteins described in our study due to the absence of the ALAVIN motif and, therefore, the U8-11 amino acid sequence was separated in a different cluster.
Highlights
Laccases are widely distributed in fungi, plants, bacteria, and insects (Rivera-Hoyos et al, 2013; Shraddha et al, 2011) and are typically extracellular monomeric glycoproteins that belong to the multicopper oxidase family (Mate & Alcalde 2017; Rodgers et al, 2010; Baldrian 2006)
For different species such as Pleurotus ostreatus which has at least 12 laccase isoenzymes (Jiao et al, 2018) and Lentinus edodes, which has 14 laccase genes identified this has been reported (Yan et al, 2019)
Phylogenetic analysis grouped strains U6-9 and U6-10 with pox1 laccase gene of P. ostreatus (AB514651 and Z34847) and lac1 laccase gene of P. sajor-caju (AF297525) in a cluster (Figure 1) and the strains U2-11, U9-1, U2-9, and U2-4 formed another cluster related to lacP83 laccase gene of P. ostreatus (Figure 1), indicating a nucleotide sequence diversification
Summary
Laccases are widely distributed in fungi, plants, bacteria, and insects (Rivera-Hoyos et al, 2013; Shraddha et al, 2011) and are typically extracellular monomeric glycoproteins that belong to the multicopper oxidase family (Mate & Alcalde 2017; Rodgers et al, 2010; Baldrian 2006). These enzymes catalyze the oxidation of phenol and polyphenol substrates with concomitant reduction of molecular oxygen to water (Cázares-García et al, 2013). The objective of this study was to identify and characterize laccase genes in silico of several basidiomycete strains
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