Decolonization of different dyes by a newly isolated white-rot fungi strain Ganoderma sp.En3 and cloning and functional analysis of its laccase gene

摘要

A laccase-producing white-rot fungi strain Ganoderma sp.En3 was newly isolated from the forest of Tzu-chin Mountain in China. Ganoderma sp.En3 had a strong ability of decolorizing four synthetic dyes, two simulated dye bath effluents and the real textile dye effluent. Induction in the activity of laccase during the decolorization process indicated that laccase played an important role in the efficient decoloriza-tion of different dyes by this fungus. Phytotoxicity study with respect to Triticum aestivum and Oryza sativa demonstrated that Ganoderma sp.En3 was able to detoxify four synthetic dyes, two simulated dye effluents and the real textile dye effluent. The laccase gene Iac-En3-1 and its corresponding full-length cDNA were then cloned and characterized from Ganoderma sp.En3. The deduced protein sequence of LAC-En3-1 contained four copper-binding conserved domains of typical laccase protein. The functionality of Iac-En3-1 gene encoding active laccase was verified by expressing this gene in the yeast Pichia pastoris successfully. The recombinant laccase produced by the yeast transformant could decolorize the synthetic dyes, simulated dye effluents and the real textile dye effluent. The ability of decolorizing different dyes was positively related to the laccase activity. In addition, the 5'-flanking sequence upstream of the start codon ATG in Iac-En3-1 gene was obtained. Many putative as-acting responsive elements were predicted in the promoter region of Iac-En3-1.