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Laccase production by pre-growth pellets of Trametes versicolor using two types of textile dyes as inducers was studied. By decoupling the enzyme production phase from the growth phase, it is possible to reduce the time and nutrients required for laccase production. At the glucose maintenance level, the effect of the nitrogen source and textile dye was analysed using response surface methodology. Ammonium chloride was used as the inorganic nitrogen source. Two types of dyes were tested: Grey Lanaset G (GLG), a metal complex dye mixture containing nitrogen; and Alizarin Red (AR), an anthraquinonic dye with no nitrogen in its chemical structure. GLG induces laccase production at a higher extent than AR. Despite the limiting conditions required for the production of laccase, enzyme production increases with increasing ammonium chloride. When AR, the N-free dye, was used as an inducer, the optimal supply of N for laccase production was 1.2 mg/(g dry cell weight x d) as ammonium chloride. The reuse of fungal pellets in the repeated-batch mode under maintenance conditions was found to be a good strategy for improving laccase production, as enzyme production increased to up to seven times the production of the first cycle. It was demonstrated that GLG can be used as an inducer and as an N source and, thus, it is possible to decolorize the dye and to induce laccase production at the same time without adding an extra N source.

Biodegradation of the Orange G azo dye by pellets of Trametes versicolor in a fluidized bioreactor operating under conditions of laccase production was studied. The percentage of decolorization obtained was 97% in batch mode and both the biomass and the broth, were colorless. In vitro degradations carried out with purified commercial laccase from Trametes versicolor demonstrated that laccase is able to degrade the dye. In spite of the high level of decolorization reached in both processes, an important difference between the fungal and enzymatic treatments was detected. At the end of the experiments carried out in vitro, a final residual color appears (different to the initial one). Consequently, measuring the yield of decolorization as a percentage of absorbance lambdamax variation is not the best indicator of the treated wastewater quality, but the analysis of the visible color spectrum makes it possible to detect changes in color. The results demonstrate that better results are obtained with fungal Orange G biodegradation because a further breakdown of the enzymatic products is achieved with the fungus.