The present study deals with the optimization of the enzymatic synthesis of n-propyl acetate using response surface methodology (RSM). The biocatalysts were prepared by physical adsorption of lipase from Thermomyces lanuginosus (TLL) on mesoporous hydrophobic supports - poly-hydroxybutyrate (PHB) and poly-methacrylate (PMA) particles. Their catalytic properties were also assayed in the hydrolysis of olive oil emulsion. The biocatalyst TLL-PMA presented the highest immobilization yield (IY, 90.6 +/- 2.8%), immobilized protein concentration (36.5 +/- 0.4 mg/g of PMA), and catalytic activity in esterification reaction. TLL-PHB was more active in the hydrolysis of olive oil emulsion by a four-fold factor (1240.5 +/- 29.2 IU/g of PHB). Maximum acid consumption percentage of 84.3% was observed after 50 min of reaction catalyzed by TLL-PMA using 2000 mM of each reactant (n-propanol and acetic acid) in heptane medium. The purified ester was confirmed by gas chromatography mass spectrometry (GC-MS) analysis. After six consecutive cycles of esterification reaction, the biocatalyst retained around 50% of its initial activity. The results show that PMA may be an interesting support to prepare active biocatalysts in the synthesis of fruit flavor ester by esterification reaction.