Recombinant expression of a laccase from Coriolopsis gallica in Pichia pastoris using a modified α-factor preproleader

•The gene from laccase of Coriolopsis gallica UAMH8260 was isolated and sequenced.•The protein was expressed in Pichia pastoris, using a modified α-factor preproleader reported to enhance laccase secretion.•The recombinant enzyme showed similar kinetic constants to those of the wild type enzyme.•The expression system is amenable for protein engineering of this versatile enzyme.

In this work we communicate the heterologous expression of a laccase from Coriolopsis gallica in Pichia pastoris. This enzyme has been reported to efficiently degrade a variety of pollutants such as industrial dyes. The expression strategy included using a previously reported modified α-factor preproleader for enhanced secretion and pAOX1, a methanol-responsive promoter. Methanol concentration, copper salts concentration and temperature were varied in order to enhance laccase expression in this heterologous system. A volumetric activity of 250 U/L was achieved after 12-day culture in Fernbach flasks. The protein was recovered from the supernatant and purified, obtaining a preparation with 90% electrophoretic purity. The catalytic constants of the recombinant enzyme are almost identical to the fungal enzyme, thus rendering this system a useful tool for protein engineering of laccase from C. gallica.