Abstract
Immobilization of lipases on hydrophobic supports at low ionic strength permits one- step purification, immobilization, hyperactivation, and stabilization of most lipases. This selective adsorption occurs because the hydrophobic surface of the supports is able to promote the interfacial activation of the lipases, yielding enzyme preparations having the open form of the lipases very strongly adsorbed on these hydrophobic supports. At low ionic strength, only proteins having large hydrophobic pockets may become adsorbed on the hydrophobic support, and the only soluble proteins are lipases, which in closed form are fairly hydrophilic, but in open form expose a very hydrophobic pocket. The result- ing biocatalysts are therefore hyperactivated, at least with hydrophobic and small substrates (because all the enzyme molecules have the open form). Moreover, the stabilization of the open form of the lipases permits very highly stabilized enzyme preparations.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Undurraga D., Markovits A., and Erazo S. (2001) Cocoa butter equivalent through enzymatic interesterification of palm oil midfraction. Process Biochem. 36, 933–939.
Plou J. F., Barandiaran M., Calvo M. V., Ballesteros A., and Pastor E. (1996) High yield production of mono and dioleylglycerol by lipase catalyzed hydrolysis of triolein. Enzyme Microb. Technol. 18, 66–71.
Jaeger K.E. and Reetz T.M. (1998) Microbial lipases from versatile tools for biotechnology. Trends Biotechnol. 16, 396–403.
Watanabe Y., Miyawaki Y., Adachi S., Nakanishi K., and Matsuno R. (2001) Continuous production of acyl mannoses by immobilized lipase using a packedbed reactor and their surfactant properties. Biochem. Eng. J. 8, 213–216.
Wong C. H. and Whitesides G. M. (1994) Enzymes in Synthetic Organic Chemistry, Tetrahedron Organic Chemistry Series, vol. 12, Pergamon Press, Oxford, UK.
Kazlauskas R. J. and Bornscheuer U. T. (1998) Biotransformations with Lipases in Biotechnology. 68–87.
Schmid R.D. and Verger R. (1998) Lipases: interfacial enzymes with attractive applications. Angew. Chem., Int. Ed. 37, 1609–1633.
Palomo J. M., Fernandez-Lorente G., Mateo C., Fuentes M., Fernandez-Lafuente R., and Guisán J. M. (2002) Modulation of the enantioselectivity of Candida antarctica B lipase via conformational engineering: kinetic resolution of (fx)-α-hydroxy-phenylacetic acid derivatives. Tetrahedron: Asymmetry. 13, 1337–1345.
Palomo J. M., Fernandez-Lorente G., Rua M. L., Guisán J. M., and Fernandez-Lafuente R. (2003) Evaluation of the lipase from Bacillus thermocatenulatus as an enantioselective biocatalyst. Tetrahedron: Asymmetry 14, 3679–3687.
Palomo J. M., Mateo C., Fernández-Lorente G., et al. (2003). Chiral resolution of (S)-5(+)-substituted-6-(5-chloropyridin-2-yl)-7-oxo-5,6-dihydropyrrolo[3,4b]pyrazine derivatives-precursors of (S)-(+)-Zopiclone, catalyzed by immobilized C andida antarctica B lipase in aqueous media. Tetrahedron: Asymmetry 14, 429–438.
Brady L., Brzozowski A. M., Derewenda Z. S., et al. (1990) A serine protease triad forms the catalytic center of a triacylglycerol lipase. Nature 343, 767–770.
Brzozowski A. M., Derewenda U., Derewenda Z. S., et al. (1991) A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex. Nature 351, 491–494.
Sarda L. and Desnuelle P. (1958) Action de la lipase pancreatique sur les esteres en emulsion. Biochim. Biophys. Acta. 30, 513–521.
Ghosh D., Wawrzak Z., Pletnev V. Z., et al. (1995) Structure of uncomplexed and linoleate-bound Candida cylindracea cholesterol esterase. Structure 3, 279–288.
GarcÃa-Alles L. F. and Gotor V. (1998) Lipase-catalyzed transesterifcation in organic media: solvent effects on equilibrium and individual rate constants. Biotechnol. Bioeng. 59, 684.694.
Bastida A., Sabuquillo P., Armisen P., Fernández-Lafuente R., Huguet J., and Guisán, J. M. (1998) A single step purification, immobilization and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol. Bioeng. 58, 486.493.
Palomo J. M., Muñoz G., Fernández-Lorente G., Mateo C., Fernández-Lafuente R., and Guisán J. M. (2002) Interfacial adsorption of lipases on very hydrophobic support (Octadecyl-Sepabeads): immobilization, hyperactivation and stabilization of the open form of lipases. J. Mol. Catal. B: Enzym. 19–20, 279–286.
Miled N., Beisson F., de Caro J., de Caro A., Arondel V., and Verger R. (2001) Interfacial catalysis by lipases. J. Mol. Catal. B: Enzym. 11, 65–171.
Palomo J. M., Segura R. L., Fernández-Lorente G., et al. (2004) Purification, Immobilization and stabilization of a lipase from Bacillus thermocatenulatus. Biotechnol. Prog. 20, 630–635.
Sabuquillo P., Reina J., Fernández-Lorente G., Guisán J. M., and Fernández-Lafuente R. (1998) Interfacial affinity chromatography of lipases: separation of different fractions by selective adsorption on supports activated with hydrophobic groups. Biochim. Biophys. Acta. 1388, 337–348.
Guisán J. M. (1988) Aldehyde gels as activated support for immobilization-stabilization of enzymes. Enzyme Microb. Technol. 10, 375–382.
Fernández-Lafuente R., Rodriguez V., and Guisan J. M. (1998) The coimmobilization of D-aminoacid oxidase and catalase enables the quantitative transformation of D-amino acids (phenylalanine) into α-ceto acids (phenylpyruvic acid). Enzyme Microb. Technol. 23, 28.33.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Palomo, J.M. et al. (2006). Purification, Immobilization, Hyperactivation, and Stabilization of Lipases by Selective Adsorption on Hydrophobic Supports. In: Guisan, J.M. (eds) Immobilization of Enzymes and Cells. Methods in Biotechnologyâ„¢, vol 22. Humana Press. https://doi.org/10.1007/978-1-59745-053-9_13
Download citation
DOI: https://doi.org/10.1007/978-1-59745-053-9_13
Publisher Name: Humana Press
Print ISBN: 978-1-58829-290-2
Online ISBN: 978-1-59745-053-9
eBook Packages: Springer Protocols