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Immobilization of Enzymes on Hetero-Functional Supports: Physical Adsorption Plus Additional Covalent Immobilization

  • Lara Trobo-Maseda
  • Alejandro H. Orrego
  • María Romero-Fernández
  • Jose M. GuisanEmail author
  • Javier Rocha-Martín
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2100)

Abstract

The immobilization of proteins on heterofunctional amino-epoxy and amino-glyoxyl supports is described in this chapter. Immobilization on both supports is performed through a two-step mechanism: in the first step, the enzyme is physically adsorbed to the support, and in the second step, the intramolecular covalent attachment between the adsorbed enzyme and the support is promoted. On the one hand, amino-epoxy supports present a ratio between amino and epoxy groups of 1:1 to allow the rapid adsorption of the enzyme and promote a strong multipoint covalent linkage. On the other hand, amino-glyoxyl supports contain the highest concentration of glyoxyl groups capable of reacting covalently with primary amino groups on the enzyme surface to promote increased rigidification. The intensity of the covalent enzyme–support interaction can be modulated by modifying the ratio between glyoxyl and amino groups of the support. These heterofunctional supports are able to immobilize and rigidify proteins through different orientations, leading to biocatalysts with different enzyme properties (activity, stability, and selectivity).

Key words

Heterofunctional epoxy supports Heterofunctional glyoxyl supports Covalent immobilization Stabilization 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Lara Trobo-Maseda
    • 1
  • Alejandro H. Orrego
    • 1
  • María Romero-Fernández
    • 1
  • Jose M. Guisan
    • 1
    Email author
  • Javier Rocha-Martín
    • 1
  1. 1.Department of BiocatalysisInstitute of Catalysis and Petrochemistry (ICP) CSIC, Campus UAMMadridSpain

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