African Journal of Biotechnology Vol. 6 (10), pp. 1248-1254, 16 May 2007   

ISSN 1684–5315 © 2007 Academic Journals        

Full Length Research Paper

Purification, characterization and application of laccase from Trametes versicolor for colour and phenolic removal of olive mill wastewater in the presence of 1-hydroxybenzotriazole

Rosana C. Minussi^1,2, Márcio A. Miranda³, José A. Silva⁴, Carmen V. Ferreira³, Hiroshi Aoyama³, Sérgio Marangoni⁴, Domenico Rotilio⁵, Gláucia M. Pastore¹ and Nelson Durán^2,6*

¹Faculdade de Engenharia de Alimentos Universidade Estadual de Campinas, Campinas - SP, Brazil

² Instituto de Química Universidade Estadual de Campinas, Campinas - SP, Brazil

³Instituto de BiologiaUniversidade Estadual de Campinas, Campinas - SP, Brazil

⁵Consorzio Mario Negri Sud, “Gennaro Paone” Environmental Health Center Santa Maria Imbaro,

Italy.

⁶Núcleo de Ciências Ambientais, Universidade de Mogi das Cruzes, Mogi das Cruzes - SP, Brazil.

*Corresponding author. E-mail: duran@iqm.unicamp.br. Tel: (0055) 19-3521-3149. Fax: (0055) 19-3521-3023.

Accepted 25 January, 2007

Laccase forms (L1 and L2) from Trametes versicolor CCT 4521 showed a molecular mass of 66 kDa and optimum temperature around 40^oC. The optimum pH (4.0 and 5.0) and K_m (28.6 and 5 µM) values using syringaldazine as substrate were found for L1 and L2, respectively. The enzymes were able to oxidize several compounds and were strongly inhibited by sodium azide, L-cysteine and dithiothreitol. The 75% of the N-terminal sequences were identical in both forms and similarities around 40 - 60% of laccases from wood-degrading fungi were observed. The use of 1-hydroxybenzotriazole as a mediator increased the compounds oxidized by laccases in olive mill wastewater.

Key words: Laccase, Trametes versicolor, 1-hydroxybenzotriazole, mediator, olive mill wastewater.