Abstract
The prospects of a new commercially available support (amino-epoxy-Sepabeads® ) for enzyme immobilization are discussed in this chapter. These supports have a layer of epoxy groups over a layer of ethylenediamine that is covalently bound to the support. Thus, the support has a high anionic exchanger power and a high number of epoxy groups. Some relevant properties of this support are (1) immobilization proceeds at low ionic strength using amino-epoxy-Sepabeads, (2) immobilization is much more rapid using amino-epoxy supports than employing conventional epoxy supports, and (3) stability of the immobilized enzyme has been found to be much higher using the new support than in preparations using the conventional ones in many cases. Therefore, this support may be a good complement to the traditional hydrophobic epoxy supports.
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References
Hartmeier W. (1985) Immobilized biocatalysts—from simple to complex sys-tems. TIBTECH 3, 149–153.
Katchalski-Katzir E. (1993) Immobilized enzymes: learning from past successes and failures. TIBTECH 11, 471–478.
Bickerstaff G. F. (1997) Immobilization of enzymes and cells: some practical considerations. In: Immobilization of Enzymes and Cells, Methods in Biotechnology (Bickerstaff G. F., ed.), vol. 1, Humana Press Inc. Totowa, NJ. 1–12.
Katchalski-Katsir E. and Kraemer D. (2000) Eupergit C, a carrier for immobiliza-tion of enzymes of industrial enzymes. J Mol. Catal. B: Enzymatic 10, 157–176.
Wheatley J. B., and Schmidt D. E. Jr. (1993) Salt induced immobilization of proteins on a high-performance liquid chromatographic epoxide afinity support. J. Chromatog. 644, 11–16.
Wheatley J. B. and Schmidt D. E. Jr. (1999) Salt induced immobilization of affinity ligands onto epoxide activated supports. J. Chromatog. A. 849, 1–12.
Mateo C., Abian O, Fernández-Lafuente R., and Guisán J. M. (2000) Increase in conformational stability of enzymes immobilized on epoxy-activated supports by favoring additional multipoint covalent attachment. Enzyme Microb. Technol. 26, 509–515.
Mateo C., Abian O., Fernández-Lorente G., Pedroche J., Fernández-Lafuente R. and Guisan J. M. (2002) Sepabeads: a novel epoxy-support for stabilization of industrial enzymes via very intense multipoint covalent attachment. Biotechnol. Progr. 18, 629–634.
Mateo C., Fernández-Lorente G., Cortes E., Garcia J. L., Fernández-Lafuente R., and Guisán J. M. (2001) One step purification, covalent immobilization and additional satabilization of poly-hys-tagged proteins using a novel heterofunctional chelate-epoxy supports. Biotechnol. Bioeng. 76, 269–277.
Melander W., Corradini D., and Hoorvath C. (1984) Salt-mediated retention of proteins in hydrophobic-interaction chromatography. Aplication of solvophobic theory. J. Chromatogr. 317, 67–85.
Smalla K., Turkova J., Coupek J., and Herman P. (1988) Influence of salts on the covalent immobilization of proteins to modified copolymers of 2-hydroxiethylmetacrylate with ethilene dimetacrylate. Biotechnol. Appl. Biochem. 10, 21–31.
Mateo C., Fernández-Lorente G., Abian O., Fernández-Lafuente R., and Guisán J. M. (2000). Multifunctional epoxy-supports: a new tool to improve the covalent immobilization of proteins. The promotion of physical adsorptions of proteins on the supports before their covalent linkage. Biomacromolecules. 1, 739–745.
Pessela B. C. C., Mateo C., Carrascosa A. V., et al. (2003). One step purifica-tion, covalent immobilization and additional stabilization of a thermophilic poly-his-tagged β-galactosidase of Thermus sp. strain t2, novel heterofunctional chelate-epoxy supports. Biomacromolecules. 4, 107–113.
Mateo C., Archelas A., Fernandez-lafuente R., Guisan J. M., and Furstoss R. (2003) Enzymatic transformations. Immobilized A. niger epoxide hydrolase as a novel biocatalytic tool for repeated-batch hydrolytic kinetic resolution of ep-oxides. Org. Biomol. Chem. 1, 2739–2743.
Grazú V., Abian O., Mateo C., Batista F., Fernández-Lafuente R., and Guisán J. M. (2003) Novel bifunctional epoxy/thiol-reactive support to immobilize thiol containing proteins by the epoxy chemistry. Biomacromolecules. 4, 1495–1501.
Torres R., Mateo C., Fernández-Lorente G., et al. (2003) A novel heterofunctional epoxy-amino Sepabeads for a new enzyme immobilization protocol: immobiliza-tion-stabilization of beta-galactosidase from Aspergillus oryzae. Biotechnol. Progr. 19, 1056–1060.
Mateo C., Torres R., Fernández-Lorente G., et al. (2003) Epoxy-amino groups: a new tool for improved immobilization of proteins by the epoxy method. Biomacro-molecules. 4, 772–777.
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Mateo, C. et al. (2006). Immobilization and Stabilization of Proteins by Multipoint Covalent Attachment on Novel Amino-Epoxy-Sepabeads®. In: Guisan, J.M. (eds) Immobilization of Enzymes and Cells. Methods in Biotechnology™, vol 22. Humana Press. https://doi.org/10.1007/978-1-59745-053-9_14
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DOI: https://doi.org/10.1007/978-1-59745-053-9_14
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