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.

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

Abbreviation

Abbreviations: ABTS, 2,2’-Azino-bis-(3-ethylbenzthiazoline- 6-sulfonic acid); SDS-PAGE, sodium dodecyl sulfate- polyacrylamide gel electrophoresis; IEF–PAGE,isoelectric focusing gel; pIs, isoelectric points.