The degradation of lindane was studied in liquid-agitated cultures using a commercial strain of the fungus Pleurotus ostreatus as the biodegrading organism. The biodegradation was accomplished with the action of extracellular oxidative enzymes, produced by the fungus to decompose woody substrates. Enzyme activities of manganese peroxidase and laccase were measured in a liquid mineral medium. An orthogonal Central Composite Design of experiments was used to construct second-order response surfaces with the fungus growth, final pH and the lindane biodegradation as optimization parameters. The initial lindane concentration, the nitrogen content, the incubation time and the temperature were used as design factors. Optimal conditions found for all these parameters will be used for the continuation of this project aiming at the bioremediation of contaminated sites with persistent organic pollutants such as lindane.