The use of enzymatic route for production
of biofuels is growing up due the mild reaction conditions that this method
provides, as well as reducing SOx emission. To reduce costs, it’s necessary to
immobilize the enzyme, making possible to use it continuously as biocatalyst.
The aim of this work was to measure the influence of the mass of support and pH
used for immobilization of commercial lipase from Candida rugosa acquired by
Sigma laboratory. The immobilization method chosen was adsorption on mesoporous
and hydrophobic support MCM 41, this has been treated with nitric acid 10% v/v
to remove any organic residue. Then, 20 ml of enzymatic solution in phosphate
buffer (pH 6.0, 7.0 and 8.0; 50 mM) and 1 g/L was placed under constant
stirring with 0.30 and 0.45 g of support. Aliquots were taken from the reaction
medium and analyzed by spectrophotometry at 10 minutes intervals. A volume of
0.2 ml of supernatant was put with 1.8 ml of substrate p-NFL at 0.18 g/L, and
the absorbance at 410 nm was analyzed. In four cases there was a sharp
reduction of supernatant’s activity at first 10 minutes, that ratifies the big
affinity of the enzyme for the support and the negative influence of pH about
the activity. Using the calibration curve, it was possible to calculate the
final activity of each immobilization batch. This work suggests the occurrence
of diffusional effects, which means that the enzyme mobility was restricted due
the excessive amount of support, and then, it lost a part of accessibility to
substrate, reflecting in not expressive activity values, and changing the state
of ionization of the components of the system.
Cite this paper
Souza, R. and Ferreira, R. (2014) Immobilization of Lipase from Candida rugosa on Mesoporous MCM 41. Journal of Biosciences and Medicines
, 69-73. doi: 10.4236/jbm.2014.24011
 Zheng, M., Dong, L., Lu, Y., Guo, P., Deng, Q., Li, W., Feng, Y. and Huang, F. (2012) Immobilization of Candida Rugosa Lipase on Magnetic Poly(Allylglycidyl Ether-co-Ethylene Glycol Dimethacrylate)Polymer Microsphere for Synthesis of Phytosterol Esters of Unsaturated Fatty Acids. Journal of MolecularCatalysis B: Enzymatic, 74, 16-23.
 Verger, R. (1997) Interfacial Activation of Lipases: Facts and Artifacts. Trends in Biotechnology, 15, 32-38.
 Bon, E.P.S., Ferrara, M.A. and Corvo, M.L. (2008) Enzimas em Biotecnologia: Produ??o, Aplica??es e Mercado. InterciênciaLtda, Rio de Janeiro.
 Yang, J.J., Ma, X.O., Zhang, Z.S., Cheng, B., Li, S. and Wang, G. (2010) Lipase Immobilized by Modification- Coupled and Adsorption-cross-Linking Methods: A Comparative Study. Biotechnology Advances, 28, 644-650.
 Reguly, J.C. (2000) Biotecnologia dos Processos Fermentativos. Vol. 3, Editora Universitário/UFPE.
 Kharrat, N., Ali, Y.B., Marzouk, S., et al. (2011) Immobilization of Rhizopusoryzae Lipase on Silica Aerogels by Adsorption: Comparison with the Free Enzyme. Process Biochemistry, 46, 1083-1089.
 Chen, G., Kuo, C., Chen, C., et al. (2011) Effect of Membranes with Various Hydrophobic/Hydrophilic Properties on Lipase Immobilized Activity and Stability. Journal of Bioscience and Bioengineering.
 Fernandez-Lafuente, R., Armisèn, P., Sabuquillo, P., Fernández-Lorente, G. and Guisán, J.M. (1998) Immobilization of Lipases by Selective Adsorption on Hydrophobic Supports. Chemistry and Physics of Lipids, 93, 185-197.
 Zhou, Z., Inayat, A., Schwieger, W. and Hartmann, M. (2012) Improved Activity and Stability of Lipase Immobilized in Cage-Like Large Pore Mesoporous Organosilicas, Micro-porous and Mesoporous Materials.
 Kandasamy, R., Kennerdy, L.J., Vidya, C., et al. (2010) Immobilization of Acidic Lipase Derived from Pseudomonas Gessardii onto Mesoporous Activated Carbon for the Hy-drolysis of Olive Oil. Journal of Molecular Catalysis B: Enzymatic, 62, 59-66. http://dx.doi.org/10.1016/j.molcatb.2009.09.004
 Kresge, C.T., Leonowics, M.E., Roth, W.J., Vartulli, J.C. and Beck, J.S. (1992) Ordered Mesoporous Molecular Sieves Synthesized by a Liquid-Crytal Template Mechanism. Nature, 359, 710. http://dx.doi.org/10.1038/359710a0