Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolorization of a recalcitrant dye.

A laccase-producing ascomycete was isolated from arid soil in Tunisia. This fungus was identified as Thielavia sp. using the phylogenetic analysis of rDNA internal transcribed spacers. The extracellular laccase produced by the fungus was purified to electrophoretic homogeneity, showing a molecular mass around 70 kDa. The enzyme had an optimum pH of 5.0 and 6.0 for ABTS and 2,6‑DMP, respectively and it showed remarkable high thermal stability, showing its optimal temperature at 70 °C (against 2,6‑DMP). It presented slight inhibiting effect by EDTA, SDS and l‑cyst although this effect was more marked by sodium azide (0.1 mM). On the other hand, it showed tolerance to up to 300 mM NaCl, retaining around 50% of its activity at 900 mM. Among the metal ions tested on TaLac1, Mn2+ showed an activating effect. Their kinetic parameters Km and kcat were 23.7 μM and 4.14 s-1 for ABTS, and 24.3 μM and 3.46 s-1 towards 2,6‑DMP. The purified enzyme displayed greater efficiency in Remazol Brilliant Blue R decolorization (90%) in absence of redox mediator, an important property for biotechnological applications.