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

Abstract

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 

References

  1. 1.
    Fessner WD (2015) Systems biocatalysis: development and engineering of cell-free “artificial metabolisms” for preparative multi-enzymatic synthesis. New Biotechnol 32:658–664CrossRefGoogle Scholar
  2. 2.
    Köhler V, Turner NJ (2014) Artificial concurrent catalytic processes involving enzymes. Chem Commun 51:450–464CrossRefGoogle Scholar
  3. 3.
    Rollin JA, Tam TK, Zhang YHP (2013) New biotechnology paradigm: cell-free biosystems for biomanufacturing. Green Chem 15:1708–1719CrossRefGoogle Scholar
  4. 4.
    Zhu Z, Kin Tam T, Sun F, You C, Percival Zhang YH (2014) A high-energy-density sugar biobattery based on a synthetic enzymatic pathway. Nat Commun 5:3026CrossRefGoogle Scholar
  5. 5.
    Jia F, Narasimhan B, Mallapragada S (2014) Materials-based strategies for multi-enzyme immobilization and co-localization: a review. Biotechnol Bioeng 111:209–222CrossRefGoogle Scholar
  6. 6.
    Schoffelen S, Van Hest JCM (2012) Multi-enzyme systems: bringing enzymes together in vitro. Soft Matter 8:1736–1746CrossRefGoogle Scholar
  7. 7.
    Huang X, Li M, Mann S (2014) Membrane-mediated cascade reactions by enzyme-polymer proteinosomes. Chem Commun 50:6278–6280CrossRefGoogle Scholar
  8. 8.
    García-García P, Rocha-Martin J, Fernandez-Lorente G, Guisan JM (2018) Co-localization of oxidase and catalase inside a porous support to improve the elimination of hydrogen peroxide: oxidation of biogenic amines by amino oxidase from Pisum sativum. Enzym Microb Technol 115:73–80CrossRefGoogle Scholar
  9. 9.
    Rocha-Martín J, Bdl R, Muñoz R, Guisán JM, López-Gallego F (2012) Rational co-immobilization of bi-enzyme cascades on porous supports and their applications in bio-redox reactions with in situ recycling of soluble cofactors. ChemCatChem 4:1279–1288CrossRefGoogle Scholar
  10. 10.
    Ji X, Su Z, Wang P, Ma G, Zhang S (2014) Polyelectrolyte doped hollow nanofibers for positional assembly of bienzyme system for cascade reaction at O/W interface. ACS Catal 4:4548–4559CrossRefGoogle Scholar
  11. 11.
    Bolivar JM, Hidalgo A, Sánchez-Ruiloba L, Berenguer J, Guisán JM, López-Gallego F (2011) Modulation of the distribution of small proteins within porous matrixes by smart-control of the immobilization rate. J Biotechnol 155:412–420CrossRefGoogle Scholar
  12. 12.
    Zheng Y-G, Yin H-H, Yu D-F, Chen X, Tang X-L, Zhang X-J, Xue Y-P, Wang Y-J, Liu Z-Q (2017) Recent advances in biotechnological applications of alcohol dehydrogenases. Appl Microbiol Biotechnol 101:987–1001CrossRefGoogle Scholar
  13. 13.
    van der Donk WA, Zhao H (2003) Recent developments in pyridine nucleotide regeneration. Curr Opin Biotechnol 14:421–426CrossRefGoogle Scholar
  14. 14.
    Bolivar JM, Cava F, Mateo C, Rocha-Martin J, Guisan JM, Berenguer J, Fernandez-Lafuente R (2008) Immobilization-stabilization of a new recombinant glutamate dehydrogenase from Thermus thermophilus. Appl Microbiol Biotechnol 80:49–58CrossRefGoogle Scholar
  15. 15.
    Rocha-Martín J, Vega D, Bolivar JM, Hidalgo A, Berenguer J, Guisán JM, López-Gallego F (2012) Characterization and further stabilization of a new anti-prelog specific alcohol dehydrogenase from Thermus thermophilus HB27 for asymmetric reduction of carbonyl compounds. Bioresour Technol 103:343–350CrossRefGoogle Scholar
  16. 16.
    Mateo C, Bolivar JM, Godoy CA, Rocha-Martin J, Pessela BC, Curiel JA, Muñoz R, Guisan JM, Fernández-Lorente G (2010) Improvement of enzyme properties with a two-step immobilizaton process on novel heterofunctional supports. Biomacromolecules 11:3112–3117CrossRefGoogle Scholar

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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