A Flexible Method for the Stable, Covalent Immobilization of Enzymes at Electrode Surfaces
| Autor: | Su Ma, Philip N. Bartlett, Marta Meneghello, Roland Ludwig, Firas A. Al-Lolage |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
chemistry.chemical_classification
Cellobiose dehydrogenase Immobilized enzyme 010405 organic chemistry Chemistry Biomolecule 010402 general chemistry 01 natural sciences Combinatorial chemistry Redox Catalysis 0104 chemical sciences chemistry.chemical_compound Electron transfer Solid-phase synthesis Electrochemistry Organic chemistry Maleimide Biosensor |
| Zdroj: | ChemElectroChem. 4:1528-1534 |
| ISSN: | 2196-0216 |
| Popis: | Stable, site-specific immobilization of redox proteins andenzymes is of interest for the development of biosensors and biofuelcells, where the long-term stability of enzymatic electrodes as wellas the possibility of controlling the orientation of the biomolecules atthe electrode surface have a great importance. Ideally, it would bedesirable to immobilise redox proteins and enzymes in a specificorientation, but still with some flexibility to optimise reaction kinetics.In this work, we establish such an approach using site-directedmutagenesis to introduce cysteine residues at specific locations onthe protein surface and the reaction between the free thiol group andmaleimide groups attached to the electrode surface to immobilisethe mutated enzymes. Using cellobiose dehydrogenase (CDH) as amodel system, carbon nanotubes electrodes were first covalentlymodified with maleimide groups following a modular approach basedon electrografting of primary amines at the carbon surface and solidphase synthesis methodology to elaborate the surface modifiedelectrode. The CDH-modified electrodes were tested for directelectron transfer (DET), showing high catalytic currents as well asexcellent long-term storage stability. The key advantage of thismethod is its great flexibility, as the main components of themodification can be independently varied to change the localenvironment at the electrode surface and a wide range of redoxproteins or enzymes can be specifically engineered to presentcysteine residues at their surface for oriented immobilisation. |
| Databáze: | OpenAIRE |
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