Background: Biological methods based on enzymatic activity measurements may offer promising perspectives for ecotoxicological risk assessment in soils contaminated by heavy metals. Because of their ecological pertinence, we retain filamentous fungi as models for the development of enzymatic methods. Aims: The present study aims to assess both selectivity and sensitivity of the response of exocellular oxidases secreted by the basidiomycete Trametes versicolor exposed to metals. Methods: We spiked with high amounts of metals pure cultures performed in liquid media to assess the selectivity of the enzymatic response. We used media exhibiting several levels of metal complexation to measure its sensitivity. Results: Laccase and manganese-peroxidase activities were highly stimulated in the presence of copper, mercury and cadmium. By contrast, lead and zinc were without significant effects on these activities. Lignin-peroxidase activity was not measured in the control cultures, but it was strongly increased after fungal exposure to copper. Assays performed with mixtures of metals, reflecting soil multiple contaminations, provided contrasted results. Then, culture media exhibiting low complexing properties led to an increased bioavailability of metals. As a consequence, we were able to measure the enzymatic responses at environmental levels of contaminants. Conclusion: Our results validate our hypothesis and allow us to develop studies in soils. We will now introduce Trametes versicolor in contaminated soils and measure the resulting enzymatic activities.