Co-Immobilization and Co-Localization of Multi-Enzyme Systems on Porous Materials

  • Alejandro H. Orrego
  • Fernando López-Gallego
  • Gloria Fernandez-Lorente
  • Jose M. Guisan
  • Javier Rocha-MartínEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2100)


The immobilization of multi-enzyme systems on solid materials is rapidly gaining interest for the construction of biocatalytic cascades with biotechnological applications in industry. The heterogenization and control of the spatial organization across porous materials of the system components are essentials to improve the performance of the process providing higher robustness, yield, and productivity. In this chapter, the co-immobilization and co-localization of a bi-enzymatic bio-redox orthogonal cascade with in situ cofactor regeneration are described. An NADH-dependent alcohol dehydrogenase catalyzes the asymmetric reduction of 2,2,2 trifluoroacetophenone using an NADH regeneration system consisting of a glutamate dehydrogenase and glutamic acid. Three different spatial organizations of the enzymes were compared in terms of cofactor-recycling efficiency. Furthermore, we demonstrated how the co-localization and uniform distribution (by controlling the enzyme immobilization rate) of the main and recycling dehydrogenases inside the same porous particle lead to enhance the cofactor-recycling efficiency of the bi-enzymatic bio-redox systems.

Key words

Immobilization Porous materials Enzymes Cofactor regeneration Co-localization Heterogeneous biocatalysis 


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Copyright information

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

Authors and Affiliations

  • Alejandro H. Orrego
    • 1
  • Fernando López-Gallego
    • 2
  • Gloria Fernandez-Lorente
    • 1
    • 3
  • Jose M. Guisan
    • 1
  • Javier Rocha-Martín
    • 1
    Email author
  1. 1.Department of BiocatalysisInstitute of Catalysis and Petrochemistry (ICP) CSIC, Campus UAMMadridSpain
  2. 2.Departamento de Química OrgánicaInstituto de Síntesis Química y Catálisis Homogénea (ISQCH) CSIC-Universidad de ZaragozaZaragozaSpain
  3. 3.Department of Biotechnology and MicrobiologyInstitute of Food Science Research (CIAL), CSIC-UAM, Campus UAMMadridSpain

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