AbstractFunalia trogii and Trametes versicolor were grown on agro byproduct wheat bran moistened with various natural moistening agents, and their effects on laccase production under solid substrate condition were investigated. Laccase was the main enzyme detected under this condition. High levels of laccase activity were obtained with solid substrate cultures moistened with olive oil mill wastewater (OOMW) or alcohol factory wastewater (vinasse). Among the cultures without inducer, T. versicolor culture was detected as a more effective laccase producer than F. trogii culture. Copper and xylidine were used as laccase inducers, and copper induced laccase production more than xylidine. The maximum laccase activity was detected as 14.18 U/mL with F. trogii grown on wheat bran moistened with 5 mM CuSO4.5H2O added 25% vinasse. Azo dye decolorization activity of the supernatants from solid substrate cultures was also tested. While the use of 0.063 U/mL F. trogii laccase in reaction solution gave 66% decolorization in a minute, it was 14% for T. versicolor. This method can be a possible alternative for valorization of lignocellulosic materials and industrial wastewaters during solid substrate fermentation and for obtaining enzyme source with very high decolorization activity.