Immobilization of Enzymes in Protein Films

  • Daniel Sánchez-deAlcázar
  • Mantas Liutkus
  • Aitziber L. CortajarenaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2100)


Heterogeneous biocatalysis usually involves the use of immobilized enzymes on solid supports. Enzymes have suitable properties in terms of efficiency and selectivity for use as immobilized catalysts. Different approaches have been developed for effective immobilization, including adsorption, covalent binding, entrapment, encapsulation, and cross-linking. Those systems offer some advantages with regard to homogeneous catalysts in solution, such as low costs, easy separation and recovery of the catalyst, reusability, and enzymatic stability. Here, we describe a new approach for the immobilization of active enzymes into homogenous films composed solely of scaffolding proteins that differs from the standard methods of enzyme immobilization on solid supports.

Key words

Enzyme Enzyme immobilization Entrapment Protein scaffolds Protein films Protein materials Heterogeneous catalysis 



Financial support for this research was obtained from the Agencia Estatal de Investigació, Spain (BIO2016-77367-R and ERACoBioTech HOMBIOCAT-PCI2018-092984), and the Basque Government (Elkartek KK-2017/00008). This work was performed under the Maria de Maeztu Units of Excellence Program from the Spanish State Research Agency– grant no. MDM-2017-0720 (CIC biomaGUNE).


  1. 1.
    Cao L (2006) Carrier-bound immobilized enzymes: principles, application and design. Wiley-VCH, WeinheimGoogle Scholar
  2. 2.
    Zdarta J, Meyer AS, Jesionowski T, Pinelo M (2018) A general overview of support materials for enzyme immobilization: characteristics, properties, practical utility. Catalysis 8(2)CrossRefGoogle Scholar
  3. 3.
    Datta S, Christena LR, Rajaram YR (2013) Enzyme immobilization: an overview on techniques and support materials. 3 Biotech 3(1):1–9CrossRefGoogle Scholar
  4. 4.
    Britton J, Majumdar S, Weiss GA (2018) Continuous flow biocatalysis. Chem Soc Rev 47(15):5891–5918CrossRefGoogle Scholar
  5. 5.
    Velasco-Lozano S, Lopez-Gallego F, Vazquez-Duhalt R, Mateos-Diaz JC, Guisan JM, Favela-Torres E (2014) Carrier-free immobilization of lipase from Candida rugosa with polyethyleneimines by carboxyl-activated cross-linking. Biomacromolecules 15(5):1896–1903CrossRefGoogle Scholar
  6. 6.
    Lopez-Gallego F, Jackson E, Betancor L (2017) Heterogeneous systems biocatalysis: the path to the fabrication of self-sufficient artificial metabolic cells. Chemistry 23(71):17841–17849CrossRefGoogle Scholar
  7. 7.
    Aranaz I, Acosta N, Heras H (2018) Enzymatic d-p-hydrophenyl glycine synthesis using chitin and chitosan as supports for biocatalyst immobilization. Biocat Biotranf 36(2):89–101CrossRefGoogle Scholar
  8. 8.
    Hyeon JE, Shin SK, Han SO (2016) Design of nanoscale enzyme complexes based on various scaffolding materials for biomass conversion and immobilization. Biotechnol J 11(11):1386–1396CrossRefGoogle Scholar
  9. 9.
    Boersma YL, Pluckthun A (2011) DARPins and other repeat protein scaffolds: advances in engineering and applications. Curr Opin Biotechnol 22(6):849–857CrossRefGoogle Scholar
  10. 10.
    Grove TZ, Cortajarena AL, Regan L (2008) Ligand binding by repeat proteins: natural and designed. Curr Opin Struct Biol 18(4):507–515CrossRefGoogle Scholar
  11. 11.
    Grove TZ, Regan L, Cortajarena AL (2013) Nanostructured functional films from engineered repeat proteins. J R Soc Interface 10(83):20130051CrossRefGoogle Scholar
  12. 12.
    Cortajarena AL, Mochrie SG, Regan L (2011) Modulating repeat protein stability: the effect of individual helix stability on the collective behavior of the ensemble. Protein Sci 20(6):1042–1047CrossRefGoogle Scholar
  13. 13.
    Kajander T, Cortajarena AL, Mochrie S, Regan L (2007) Structure and stability of designed TPR protein superhelices: unusual crystal packing and implications for natural TPR proteins. Acta Crystallogr D Biol Crystallogr 63:800–811CrossRefGoogle Scholar
  14. 14.
    Mejias SH, Aires A, Couleaud P, Cortajarena AL (2016) Designed repeat proteins as building blocks for nanofabrication. Adv Exp Med Biol 940:61–81CrossRefGoogle Scholar
  15. 15.
    Mejías SH, López-Andarias J, Sakurai T, Yoneda S, Erazo KP, Seki S, Atienza C, Martín N, Cortajarena AL (2016) Repeat protein scaffolds: ordering photo- and electroactive molecules in solution and solid state. Chem Sci 7:4842–4847CrossRefGoogle Scholar
  16. 16.
    López-Andarias J, Mejías SH, Sakurai T, Matsuda W, Seki S, Feixas F, Osuna S, Atienza C, Martín N, Cortajarena AL (2017) Toward bioelectronic nanomaterials: photoconductivity in protein-porphyrin hybrids wrapped around SWCNT. Adv Funct Materials 28(24):1704031CrossRefGoogle Scholar
  17. 17.
    Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4(11):2411–2423CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Daniel Sánchez-deAlcázar
    • 1
  • Mantas Liutkus
    • 1
  • Aitziber L. Cortajarena
    • 1
    • 2
    Email author
  1. 1.CIC biomaGUNEDonostia-San SebastianSpain
  2. 2.Ikerbasque, Basque Foundation for ScienceBilbaoSpain

Personalised recommendations