Figure 1.- Effect of detergent on the activity of Lecitase lipase against p-NPB. Experimental condictions are describeb in methods. Relative activity consideres 100 the lipase activity in absence of detergent. Circles: Triton X-100, squares: CTAB.

Figure 2.- Immobilization course of Lecitase lipase on octyl-agarose support. Supernatant of the immobilization suspension (triangles),supernatant of the blank suspension (circles). The experiments were performed as described in materials and methods.

Figure 3.- Desorption of Lecitase lipase adsorbed on octyl-agarose by progressive addition of CTAB. Experiments were performed at 25 ◦C and pH 7.

Figure 4.- Effect of the CTAB concentration on the activity againts p-NPB of differnets immobilized preparations of Lecitase. Relative activity considers 100 the lipase activity in absence of detergent. Rhombus: BrCN-lecitase, squares: PEI-lecitase.

Figura 5. - Thermostability of the differents preparation of lecitase Figura 5.- Thermostability of the differents preparation of lecitase. Inactivation was carried out in 25 mM sodium phosphate buffer, pH 7 and 45ºC. Squares:BrCN-lecitase; triangles: Octyl-lecitase; circles: PEI-lecitase.

Figure 6. - Thermostability of the lecitase Figure 6.- Thermostability of the lecitase. Inactivation was carried out in 25 mM sodium Phosphate buffer, pH 5 and 55ºC. Squares:Lecitase soluble; triangles: PEI-Lecitase.

Figure 7. - Thermostability of the differents preparation of lecitase Figure 7.- Thermostability of the differents preparation of lecitase. Inactivation was carried out in 25 mM sodium phosphate buffer, pH 5 and 45ºC. Squares:BrCN-lecitase; triangles: PEI-lecitase; circles: Soluble lecitse; rhombus: Octyl-lecitase.

INACTIVACION DE DERIVADOS DE LECITASE EN PRESENCIA DE 30% DE DISOLVENTES A 25ºC Y pH 7

Table 1. Hydrolytic resolution of (±)-Hydroxy-phenylacetic acid methyl ester catalyzed by Lecitase immobilized at 25°C and different pH. Deivate Condictions Act. ee E Isomer OCTYL pH 5 0.018 0.52 3.35 R pH 7 0.083 0.46 3.02 PEI 0.033 0.75 7.47 0.058 0.51 3.34 0.01%CTAB 2.91 BrCN 0.001 0.37 2.21 0.0016 0.69 5.74 0.0017 0.61 4.35 Ee=enantiomeric excess of the product at 15% of conversion; E=value of enantioselectivity.

Derivate Condictions Act. ee E Isomer OCTYL pH 5 0.0023 0.99 >100 R Table 2. Enantioselectivity of immobilized preparations of Lecitase-catalyzed hydrolysis of 2-O-butyryl-2-phenylacetic acid at different condictions. Derivate Condictions Act. ee E Isomer OCTYL pH 5 0.0023 0.99 >100 R pH 7 0.0018* 0.84 12 PEI 0.0011 0.75 8 0.0004* 0.29 2 0.01%CTAB BrCN 0.0005 0.92 26 S 0.0014* 0.13 1.3 * Actividad medida en UI / hora , el resto son actividades por minuto. Ee=enantiomeric excess of the product at 15% of conversion; E=value of enantioselectivity.