A new laccase from the filamentous fungus Podospora anserina has been isolated and identified. The 73 kDa protein containing 4 coppers, truncated from its first 31 amino acids, was successfully overexpressed in Pichia pastoris and purified in one step with a yield of 48% and a specific activity of 644 U mg(-1). The kinetic parameters, k(cat) and K-M, determined at 37 degrees C and optimal pH are 1372 s(-1) and 307 mu M for ABTS and, 1.29 s(-1) and 10.9 mu M, for syringaldazine (SGZ). Unlike other laccases, the new protein displays a better thermostability, with a half life > 400 min at 37 degrees C, is less sensitive to chloride and more stable at pH 7. Even though, the new 566 amino-acid enzyme displays a large homology with Bilirubin oxidase (BOD) from Myrothecium verrucaria (58%) and exhibits the four histidine rich domains consensus sequences of BODs, the new enzyme is not able to oxidize neither conjugated nor unconjugated bilirubin. (c) 2012 Elsevier Inc. All rights reserved.