Improvement of laccase production by Trametes versicolor was made by employing different operational strategies. In the cell growth medium, various glucose concentrations were compared for improving laccase production. A clear and significant stimulation of enzyme production under carbon limitation was obtained. Copper, 2,5-xylidine, and a phenolic mixture were also used as laccase inducers. A cooperative effect between the inducers on laccase production was identified. Mixtures of inducers produced higher laccase activities, reaching values of 5500 U dm−3. Further productivity enhancement can be obtained using the inducers along with the carbon limitation strategy. It is shown that low laccase concentrations are obtained by a primary metabolism of T versicolor, and that phenolic compounds and carbon limitation induce a secondary metabolism, providing higher laccase concentrations. A mathematical model for laccase production based on a direct experimental measure of biomass, along with substrate consumption and enzymatic activity over time is proposed for non-homogeneous fermentations of T versicolor. Copyright © 2005 Society of Chemical Industry