Purification and characterization of three laccase isozymes from the white rot fungus <i>Trametes</i> sp. HS-03

Abstract

Three laccase isozymes (LacI, LacII and LacIII) were isolated from the culture supernatant solution of Trametes sp. HS-03. Diethylaminoethyl (DEAE)-sepharose fast flow anion exchange chromatography and Sephadex G-100 size-exclusion chromatography was performed to achieve electrophoretic homogeneity. The molecular masses (64.2, 60.7 and 38.9 kDa), isoelectric points [pIs (7.3, 4.7 and 3.5), and N-terminal amino acid sequences (G-I-G-P-V, A-I-G-P-T and S-I-G-P-V) were found to be different for the three laccase isozymes. LacI and II have similar thermostability, while LacIII showed better thermostability. LacIII also showed optimal activity at 80°C, with a half-life of 125 min at 70°C. The pI-value of LacI and the molecular mass of LacIII differ significantly from previously described fungal laccases.

Keywords: Trametes sp. HS-03, laccase isozymes, purification, characterization