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Catalysts
Volume 10
Issue 6
10.3390/catal10060676
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Article

High Stabilization of Enzymes Immobilized on Rigid Hydrophobic Glyoxyl-Supports: Generation of Hydrophilic Environments on Support Surfaces

by
Alejandro H. Orrego
1,
María Romero-Fernández
1,
María del Carmen Millán-Linares
2,
Justo Pedroche
2,
José M. Guisán
1,* and
Javier Rocha-Martin
1,*
1
Department of Biocatalysis, Institute of Catalysis and Petrochemistry (ICP) CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain
2
Group of Plant Proteins, Instituto de la Grasa CSIC, Carretera Utrera Km 1, 41013 Seville, Spain
*
Authors to whom correspondence should be addressed.
Catalysts 2020, 10(6), 676; https://doi.org/10.3390/catal10060676
Submission received: 29 May 2020 / Revised: 9 June 2020 / Accepted: 15 June 2020 / Published: 16 June 2020
(This article belongs to the Special Issue Advances in the Design and Characterization of Heterogeneous Biocatalysts)
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Abstract

Very rigid supports are useful for enzyme immobilization to design continuous flow reactors and/or to work in non-conventional media. Among them, epoxy-methacrylic supports are easily functionalized with glyoxyl groups, which makes them ideal candidates for enzyme stabilization via multipoint covalent immobilization. However, these supports present highly hydrophobic surfaces, which might promote very undesirable effects on enzyme activity and/or stability. The hydrophilization of the support surface after multipoint enzyme immobilization is proposed here as an alternative to reduce these undesirable effects. The remaining aldehyde groups on the support are modified with aminated hydrophilic small molecules (glycine, lysine or aspartic acid) in the presence of 2-picoline borane. The penicillin G acylase from Escherichia coli (PGA) and alcohol dehydrogenase from Thermus thermophilus HB27 (ADH2) were immobilized on glyoxyl-functionalized agarose, Relizyme and Relisorb. Despite the similar density of aldehyde groups displayed by functionalized supports, their stabilization effects on immobilized enzymes were quite different: up to 300-fold lower by hydrophobic supports than by highly hydrophilic glyoxyl-agarose. A dramatic increase in the protein stabilities was shown when a hydrophilization treatment of the hydrophobic support surface was done. The PGA immobilized on the glyoxyl-Relisorb hydrophilized with aspartic acid becomes 280-fold more stable than without any treatment, and it is even more stable than the PGA immobilized on the glyoxyl agarose.
Keywords: protein stabilization; protein immobilization; 2-picoline borane; methacrylic support; microenvironment; biocatalysis; support hydrophilization protein stabilization; protein immobilization; 2-picoline borane; methacrylic support; microenvironment; biocatalysis; support hydrophilization

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MDPI and ACS Style

H. Orrego, A.; Romero-Fernández, M.; Millán-Linares, M.d.C.; Pedroche, J.; Guisán, J.M.; Rocha-Martin, J. High Stabilization of Enzymes Immobilized on Rigid Hydrophobic Glyoxyl-Supports: Generation of Hydrophilic Environments on Support Surfaces. Catalysts 2020, 10, 676. https://doi.org/10.3390/catal10060676

AMA Style

H. Orrego A, Romero-Fernández M, Millán-Linares MdC, Pedroche J, Guisán JM, Rocha-Martin J. High Stabilization of Enzymes Immobilized on Rigid Hydrophobic Glyoxyl-Supports: Generation of Hydrophilic Environments on Support Surfaces. Catalysts. 2020; 10(6):676. https://doi.org/10.3390/catal10060676

Chicago/Turabian Style

H. Orrego, Alejandro, María Romero-Fernández, María del Carmen Millán-Linares, Justo Pedroche, José M. Guisán, and Javier Rocha-Martin. 2020. "High Stabilization of Enzymes Immobilized on Rigid Hydrophobic Glyoxyl-Supports: Generation of Hydrophilic Environments on Support Surfaces" Catalysts 10, no. 6: 676. https://doi.org/10.3390/catal10060676

APA Style

H. Orrego, A., Romero-Fernández, M., Millán-Linares, M. d. C., Pedroche, J., Guisán, J. M., & Rocha-Martin, J. (2020). High Stabilization of Enzymes Immobilized on Rigid Hydrophobic Glyoxyl-Supports: Generation of Hydrophilic Environments on Support Surfaces. Catalysts, 10(6), 676. https://doi.org/10.3390/catal10060676

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