This work reports the application of a lipase in the glycerolysis of olive oil for the production of monoacylglycerols (MAG). For this purpose, the enzymatic glycerolysis of olive oil with an immobilized lipase (Novozym 435) was accomplished using sodium (bis-2-ethyl-hexyl) sulfosuccinate (Aerosol-OT or AOT) as surfactant in a solvent-free system. A sequential strategy was used applying two experimental designs to optimize the MAG production. In general, it was observed that the temperature had a negligible effect on MAG conversion while the enzyme and AOT concentrations present a positive significant effect (p ,0.05) and the stirring rate a negative one. An empirical model was then built so as to assess the effects of process variables on the reaction conversion. Afterwards, the operating conditions that optimized MAG production were established as follows: temperature of 70 7C, stirring rate of 600 rpm, 16 wt-% of AOT and 7.5 wt-% of enzyme, leading to a reaction conversion as high as 55%.