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Article Dans Une Revue Langmuir Année : 2014

Enzyme Immobilization on Silane-Modified Surface through Short Linkers: Fate of Interfacial Phases and Impact on Catalytic Activity

Résumé

We investigated the mechanism of enzyme immobilization on silanized surfaces through coupling agents (cross-linkers) in order to understand the role of these molecules on interfacial processes and their effect on catalytic activity. To this end, we used a model multimeric enzyme (G6PDH) and several cross-linking molecules with different chemical properties, including the nature of the end-group (-NCO, -NCS, -CHO), the connecting chain (aliphatic vs aromatic), and geometrical constraints (meta vs para-disubstituted aromatics). There did not seem to be radical differences in the mechanism of enzyme adsorption according to the linker used as judged from QCM-D, except that in the case of DIC (1,4-phenylene diisocyanate) the adsorption occurred more rapidly. In contrast, the nature of the cross-linker exerted a strong influence on the amount of enzyme immobilized as estimated from XPS, and more unexpectedly on the stability of the underlying silane layer. DIC, PDC (1,4-phenylene diisothiocyanate), or GA (glutaraldehyde) allowed successful enzyme immobilization. When the geometry of the linker was changed from 1,4-phenylene diisothiocyanate to 1,3-phenylene diisothiocyanate (MDC), the silane layer was subjected to degradation, upon enzyme adsorption, and the amount of immobilized molecules was significantly lowered. TE (terephtalaldehyde) and direct enzyme deposition without cross-linker were similar to MDC. The organization of immobilized enzymes also depended on the immobilization procedure, as different degrees of aggregation were observed by AFM. A correlation between the size of the aggregates and the catalytic properties of the enzyme was established, suggesting that aggregation may enhance the thermostability of the multimeric enzyme, probably through a compaction of the 3D structure.

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Chimie
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Dates et versions

hal-01121650 , version 1 (02-03-2015)

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Nesrine Aissaoui, Latifa Bergaoui, Souhir Boujday, Jean-François Lambert, Christophe Méthivier, et al.. Enzyme Immobilization on Silane-Modified Surface through Short Linkers: Fate of Interfacial Phases and Impact on Catalytic Activity. Langmuir, 2014, 30 (14), pp.4066-77. ⟨10.1021/la404935q⟩. ⟨hal-01121650⟩
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