Enantiomerically pure compounds is mainly made by the standard resolution of racemic mixtures, which can only achieve a maximum of 50% yield. As a solution to this problem the asymmetric catalysis can obtain >99% yields in a single reaction step. Chiral 3-arylglutaric acid derivatives are important building blocks for the synthesis of several biologically active compounds that possess a chiral glutaric fragment in their structures, which is essential for their activity1. In this work, we seek to improve the enantiomeric excess obtained for this type of molecules, studying the effect of protein load in immobilized enzymes. The asymmetric hydrolysis of dimethyl 3-phenylglutarate catalyzed by an immobilized preparation of Geobacillus Thermocatenulatus lipase (BTL2) has been studied. Assays were carried out using different loads of protein. The commercial support Agarosa 6BCL was converted to heterofunctional support and used for the immobilization of BTL2 in two steps, first physical adsorption and then multipoint covalent attachment2. The lipase immobilized with maximum protein load showed the most selective catalyst for the hydrolysis dimethyl 3-phenylglutarate at pH 7 and 25 °C producing the (S)-monoester with an activity value of 62 UI/g support (Figure 1) and E value of 9,5 (%ee=80, 9) (Figure 2). The results clearly show that increasing the loaded protein on the support, improves the catalyst selectivity. The best result reached was for the saturated catalyst, i.e. with the greatest possible amount of loaded protein. Probably due to this type of immobilization in two steps, a greater amount of protein in the support promoted a particular orientation to improve their selectivity and activity. It is proposed as the next step in this work, to use a labeled protein to evaluate its location in the support.