Covalent immobilization of pure lipases A and B from Candida rugosa

Covalent immobilization on agarose and SiO2 of pure lipase A (LIP.A) and B (LIP.B) from C. rugosa is described. The results obtained are compared with the data obtained in the covalent binding of commercial lipase (CL) to the same supports. The immobilization of LIP.A and LIP.B on agarose affords more stable biocatalysts than on SiO2. Kinetic studies of all these lipases and derivatives in the hydrolysis of (R)-(+) and (S)-(−) methyl 2-chloropropionates were performed and their enantiomeric ratios (E = (VmaxKm)fast(VmaxKm)slow) calculated. The results show that (i) purification of the commercial lipase increased the E value 2.5-fold; (ii) both isoenzymes have similar E values, and (iii) in general, the E value increases with the immobilization process.