Solid-state fermentation requires the development of more efficient cultivation systems for its industrial application. The objective of this work was to evaluate the effect of aeration regime on the production of biomass and several lignocellulolytic enzymes (laccase, manganese peroxidase, endoxylanase, β-glucosidase, and total cellulases) by Trametes versicolor in a novel fixed-bed solid-state pilot-scale bioreactor. Three regimes were assessed: natural convection, low aeration level (10 min every 6 h at 10 L/h air flowrate), and high aeration level (1 h every 6 h at 10 L/h air flowrate). The mushroom was grown on a medium based on lignocellulosic residues. The design of the bioreactor, as well as the control of aeration, humidity, and temperature of the beds, allowed T. versicolor to grow properly on the medium, reaching a maximum biomass production of 204.7 mg/g dry solid (ds). The influence that aeration regime had on the production of lignocellulolytic enzymes was determined. Low level of forced aeration favored obtaining the highest titers of laccase (6.37 U/g ds) compared to natural convection and high aeration level. The greatest lignin degradation was also verified for low aeration regime. For the first time, pilot scale cultivation of T. versicolor was reported in a fixed-bed bioreactor.