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Multi-Point Covalent Immobilization of Enzymes on Glyoxyl Agarose with Minimal Physico-Chemical Modification: Stabilization of Industrial Enzymes

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Immobilization of Enzymes and Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2100))

Abstract

Stabilization of enzymes via immobilization techniques is a valuable approach in order to convert a necessary protocol (immobilization) into a very interesting tool to improve key enzyme properties (stabilization). Multipoint covalent attachment of each immobilized enzyme molecule may promote a very interesting stabilizing effect. The relative distances among all enzyme residues involved in immobilization have to remain unaltered during any conformational change induced by any distorting agent. Amino groups are very interesting nucleophiles placed on protein surfaces. The immobilization of enzyme through the region having the highest amount of amino groups (Lys residues) is key for a successful stabilization. Glyoxyl groups are small aliphatic aldehydes that form very unstable Schiff’s bases with amino groups, and they do not seem to be useful for enzyme immobilization at neutral pH. However, under alkaline conditions, glyoxyl supports are able to immobilize enzymes via a first multipoint covalent immobilization through the region having the highest amount of lysine groups. Activation of supports with a high surface density of glyoxyl groups and the performance of very intense enzyme–support multipoint covalent attachments are here described.

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Author information

Authors and Affiliations

  1. Department of Biocatalysis, Institute of Catalysis and Petrochemistry (ICP) CSIC, Campus UAM, Madrid, Spain

    Fernando López-Gallego, Gloria Fernandez-Lorente, Javier Rocha-Martín, Juan M. Bolivar, Cesar Mateo & Jose M. Guisan

  2. Department of Biotechnology and Microbiology, Institute of Food Science Research (CIAL), CSIC-UAM, Campus UAM, Madrid, Spain

    Gloria Fernandez-Lorente

  3. Departamento de Química Orgánica, Instituto de Síntesis Química y Catálisis Homogénea (ISQCH) CSIC-Universidad de Zaragoza, Zaragoza, Spain

    Fernando López-Gallego

Authors
  1. Fernando López-Gallego
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  2. Gloria Fernandez-Lorente
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  3. Javier Rocha-Martín
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  4. Juan M. Bolivar
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  5. Cesar Mateo
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  6. Jose M. Guisan
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Corresponding author

Correspondence to Jose M. Guisan .

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Editors and Affiliations

  1. CSIC Campus Cantoblanco, Instituto de Catalisis, Madrid, Spain

    Jose M. Guisan

  2. Institute of Biotech and Biochemical Engineering, Graz University of Technology, Graz, Austria

    Juan M. Bolivar

  3. CIC biomaGUNE, Donostia-San Sebastian, Spain

    Fernando López-Gallego

  4. Campus UAM-CSIC, Institute of Catatalysis, Madrid, Spain

    Javier Rocha-Martín

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López-Gallego, F., Fernandez-Lorente, G., Rocha-Martín, J., Bolivar, J.M., Mateo, C., Guisan, J.M. (2020). Multi-Point Covalent Immobilization of Enzymes on Glyoxyl Agarose with Minimal Physico-Chemical Modification: Stabilization of Industrial Enzymes. In: Guisan, J., Bolivar, J., López-Gallego, F., Rocha-Martín, J. (eds) Immobilization of Enzymes and Cells. Methods in Molecular Biology, vol 2100. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0215-7_5

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  • DOI: https://doi.org/10.1007/978-1-0716-0215-7_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0214-0

  • Online ISBN: 978-1-0716-0215-7

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