Abstract
A selection of the best combination of adequate immobilization support and efficient immobilization method is still a key requirement for successful application of immobilized enzymes on the industrial level. Eupergit® supports exhibit good mechanical and chemical properties and allow establishment of satisfactory hydrodynamic regime in enzyme reactors. This is a good recommendation for their wide application in enzyme immobilization after finding the most favorable immobilization method. Methods for enzyme immobilization that have been previously reported as efficient, considering the obtained activity of immobilized enzyme are presented: direct binding to polymers via their epoxy groups, binding to polymers via a spacer made from ethylene diamine/glutaraldehyde, and coupling the periodate-oxidized sugar moieties of the enzymes to the polymer beads.
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References
Sheldon, R. A. (2007) Enzyme immobilization: The quest for optimum performance. Adv. Synth. Catal. 349, 1289–1307.
Balcao V. M., Paiva A. L., and Malcata X. F. (1996) Bioreactors with immobilized lipases: State of the art. Enzyme Microb. Technol. 18, 392–416.
Malcata, X., Reyes, H., Garcia, H., Hill, C., and Amundson, C. (1992). Kinetics and mechanisms of reactions catalyzed by immobilized lipases. Enzyme Microb. Technol. 14, 426–446.
Katchalski-Katzir, E. and Kraemer, D. M. (2000) Eupergit® C, a carrier for immobilization of enzymes of industrial potential. J. Mol. Catal. B Enzym. 10, 157–176.
Hernaiz, M. J. and Crout, D. H. G. (2000) Immobilization/stabilization on Eupergit C of the β-galactosidase from B. circulans and an α-galactosidase from Aspergillus oryzae. Enzyme Microb. Technol. 27, 26–32.
Rocchietti, S., San Vicente Urrutia, A., Pregnolato, M., Tagliani, A., Guissan, J. M., Fernandez-Lafuente, R., and Terreni, M. (2002) Influence of the enzyme derivative preparation and substrate structure on the enantioselectivity of penicillin G acylase. Enzyme Microb. Technol. 31, 88–93.
Wirz, B., Barner, R., and Huebscher, J. (1993) Facile chemoenzymic preparation of enantiomerically pure 2-methylglycerol derivatives as versatile trifunctional C4-synthons. J. Am. Chem. Soc. 58, 3980–3984.
Hilal N., Kochkodan V., Nigmatullin R., Goncharuk V., and Al-Khatib L. (2006) Lipase-immobilized biocatalytic membranes for enzymatic esterification: Comparison of various approaches to membrane preparation. J. Membr. Sci. 268, 198–207.
Mateo C., Abian O., Bernedo M., Cuenca E., Fuentes M., Fernandez-Lorente G., Palomo J. M., Grazu V., Pessela B., Giacomini C., Irazoqui G., Villarino A., Ovsejevi K., Batista-Viera F., Fernandez-Lorente R., and Guisan J. M. (2005) Some special features of glyoxyl supports to immobilize proteins. Enzyme Microb. Technol. 37, 456–462.
Kneževic´, Z., Milosavic´, N., Bezbradica, D., Jakovljevic´, Ž., and Prodanovic´, R. (2006) Immobilization of lipase from Candida rugosa on Eupergit® C supports by covalent attachment. Biochem. Eng. J. 30, 269–278.
Licia M. P., Cintia M. R., Mario D. B., and Guillermo R. C. (2006) Catalytic Properties of Lipase Extracts from Aspergillus niger. Food Technol. Biotechnol. 44, 247–252.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951) Protein measurement with Folin-phanol reagent. J. Biol. Chem. 193, 265–275.
Bradford, M. M. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
Bezbradica, D., Mijin, D., Mihailović, M., and Kneževic´-Jugovic´, Z. (2009) Microwave-assisted lipase immobilization of lipase from Candida rugosa on Eupergit® supports. J. Chem. Technol. Biotechnol. 84, 1642–164810.1002/jctb.2222.
Fernandez-Lafuente, R., Rosell, C. M., Rodrıguez, V., Santana, C., Soler, G., Bastida, A., and Guisan, J. M. (1993) Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method. Enzyme Microb. Technol. 15, 546–550.
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This work was supported by the Ministry of Science and Technological Development of the Republic of Serbia (Project No TR-20064).
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Knežević-Jugović, Z.D., Bezbradica, D.I., Mijin, D.Ž., Antov, M.G. (2011). The Immobilization of Enzyme on Eupergit® Supports by Covalent Attachment. In: Minteer, S. (eds) Enzyme Stabilization and Immobilization. Methods in Molecular Biology, vol 679. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-895-9_9
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DOI: https://doi.org/10.1007/978-1-60761-895-9_9
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