Selection Of Stabilizing Additive For Lipase Immobilization On Controlled Pore Silica By Factorial Design

Candida rugosa lipase was covalently immobilized on silanized controlled pore silica (CPS) previously activated with glutaraldehyde in the presence of several additives to improve the performance of the immobilized form in long-term operation. Proteins (albumin and lecithin) and organic molecules (β-cyclodextrin and polyethylene glycol [PEG]-1500) were added during the immobilization procedure, and their effects are reported and compared to the behavior of the immobilized biocatalyst in the absence (lacking) of additive. The selection of the most efficient additive at different lipase loadings (150-450 U/g of dry support) was performed by experimental design. Two 22 full factorial designs with two repetitions at the center point were employed to evaluate the immobilization yield. A better stabilizing effect was found when small amounts of albumin or PEG-1500 were added simultaneously to the lipase onto the support. The catalytic activity had a maximum (193 U/mg) for lipase loading of 150 U/g of dry support using PEG-1500 as the stabilizing additive. This immobilized system was used to perform esterification reactions under repeated batch cycles (for the synthesis of butyl butyrate as a model). The half-life of the lipase immobilized on CPS in the presence of PEG-1500 was found to increase fivefold compared with the control (immobilized lipase on CPS without additive).91-93703718Faber, K., (1997) Biotransformation in Organic Chemistry: A Textbook, 3rd Ed., , Springer-Verlag, BerlinYahya, A.R.M., Anderson, W.A., Moo-Young, M., (1998) Enzyme Microb. Technol., 23, pp. 438-450Reslow, M., Adlercreutz, P., Mattiason, B., (1988) Eur. J. Biochem., 172, pp. 573-578Balcão, V.M., Paiva, A.L., Malcata, F.X., (1996) Enzyme Microb. Technol., 18, pp. 392-416Bosley, J.A., Peilow, A.D., (1997) J. Am. Oil Chem. Soc., 74, pp. 107-111Soares, C.M.F., De Castro, H.F., De Moraes, F.F., Zanin, G.M., (1999) Appl. Biochem. Biotechnol., 77-79, pp. 745-758Reetz, M.T., Zonta, A., Simpelkamp, J., (1996) Biotechnol. Bioeng., 49, pp. 527-534Wehtje, E., Adlercreutz, P., Mattiasson, B., (1993) Biotechnol. Bioeng., 41, pp. 171-178Rocha, J.M.S., Gil, M.H., Garcia, F.A.P., (1998) J. Biotechnol., 66, pp. 61-67Triantafyllou, A.O., Wehtje, E., Adlercreutz, P., Mattiasson, B., (1995) Biotechnol. Bioeng., 45, pp. 406-414Triantafyllou, A.O., Wehtje, E., Adlercreutz, P., Mattiasson, B., (1997) Biotechnol. Bioeng., 54, pp. 67-76Box, G.E.P., Hunter, W.G., Hunter, J.S., (1978) Statistics for Experimenters: An Introduction to Design, Data Analysis and Model Building, , Wiley & Sons, New YorkSzejtli, J., (1988) Cyclodextrin Technology, , Kluwer Academic, BostonBradford, M.M.A., (1976) Anal. Biochem., 72, pp. 248-254De Castro, H.F., Anderson, W.A., Legge, R.L., Moo-Young, M., (1992) Indian J. Chem., 31, pp. 891-89