The lipase produced by the Aspergillus niger strain AC-S4 has been widely studied due to its enantioselectivity for racemic mixtures. This study aimed to optimize the production of this enzyme using statistical methodology. Initially a Plackett-Burman (PB) design was used to evaluate the effects of the culture medium components and the culture conditions. Twelve factors were screened: water content, glucose, yeast extract, peptone, olive oil, temperature, $NaH_2PO_4,;KH_2PO_4,;MgSO_4{cdot}7H_2O,;CaCl_2$, NaCl, and $MnSO_4$. The screening showed that the significant factors were water content, glucose, yeast extract, peptone, $NaH_2PO_4$, and $NH_2PO_4$, which were optimized using response surface methodology (RSM) and a mathematical model obtained to explain the behavioral process. The best lipase activity was attained using the following conditions: water content (20%), glucose (4.8%), yeast extract (4.0%), and $NaH_2PO_4$ (4.0%). The predicted lipase activity was 33.03 U/ml and the experimental data confirmed the validity of the model. The enzymatic activity was expressed as ${mu}moles$ of oleic acid released per minute of reaction (${mu}mol$/min).