A laboratory study was conducted to evaluate the response of soil enzyme activities (acid and alkaline phosphatase, beta-glucosidase, arylsulfatase, urease and dehydrogenase) to different levels of trace elements pollution in soils representative of the area affected by the pyrite sludge mining spill of Aznalcóllar (Guadiamar basin, SW Spain). Three uncontaminated soils from the study area were mixed with different loads of pyrite sludge to resemble field conditions and criteria applied for reclamation practices following the pollution incident: 0% ("reference" or background level), 1.3% ("attention level", further monitoring required), 4% ("intervention level", further cleaning and liming required) and 13% (ten times the "attention level"). Enzyme activities were analysed 4, 7, 14, 21, 34 and 92 days after pollutant addition and those measured after 92 days were used to calculate the ecological dose value (ED50). Soil enzyme activities and pH decreased after the pyrite sludge addition with respect to the "reference level" (0% pyrite sludge), whereas soil bioavailable (DTPA-extractable) trace elements concentration increased. Arylsulfatase, beta-glucosidase and phosphatase activities were reduced by more than 50% at 1.3% pyrite sludge dose. Arylsulfasate was the most sensitive soil enzyme (in average, ED50=0.99), whereas urease activity showed the lowest inhibition (in average, ED50=7.87) after pyrite sludge addition. Our results showed that the ecological dose concept, applied to enzyme activities, was satisfactory to quantify the effect of a multi-metalic pollutant (pyrite sludge) on soil functionality, and would provide manageable data to establish permissible limits of trace elements in polluted soils. Additionally, we evaluate the recovery of enzyme activities after addition of sugar-beet lime (calcium carbonate) to each experimentally polluted soil. The amount of lime added to each soil was enough to raise the pH to the original value (equal to control soil), resembling field remediation practices. After lime amendment, soil recovery was still incomplete in terms of bioavailable trace elements. However, the recovery of soil enzyme activities varied widely, ranging from 0 to 100% depending on soil type and the specific enzyme.