The use of lipases as biocatalysts has attracted much attention recently, but industrial application has been hindered mainly due to the high production cost, determined by the production yield and enzyme stability. Immobilization of biocatalysts in inert supports can ensure their use for several batches, making possible to build cost-effective processes. The objective of this work was to investigate the influence of immobilization time and enzyme to support mass ratio on the yield of immobilization and esterification activity of porcine pancreatic lipase. Higher immobilization yields (38.2%) were obtained for pillared montmorillonite, 120 min of immobilization and enzyme to support mass ratio of 2:0.5. The highest esterification activity (1403 U/g) was achieved using the montmorillonite, after 180 min of immobilization and enzyme to support ratio of 2:1. The characterization of the supports, free and immobilized enzyme on different supports made possible to elucidate the immobilization results through the knowledge of support features.