Electroreduction of laccase covalently bound to organothiol monolayers on gold electrodes

Abstract

Cerrena unicolor laccase was immobilized on the gold electrode by covalent bonding to self-assembled monolayers of mercaptoundecanoic or mercaptopropionic acids. STM images of immobilized laccase proved high population of the laccase molecules on the monolayer modified electrode. The SERS experiments in concert with resonance Raman effect confirmed that the structure at the “blue” copper site of the immobilized protein remained intact. The accessibility of individual copper sites for electron exchange with the gold electrode surface was investigated by voltammetry. The electrode behavior of laccase is different in the presence and absence of oxygen, showing that the immobilized enzyme is reactive towards oxygen. Addition of two common mediators improved the electrical connectivity of the enzyme with the electrode, increased the catalytic efficiency of immobilized laccase and switched the onset of catalytic current to the potentials of the mediator. Immobilization of laccase on well-organized mercaptoundecanoic acid separates efficiently the enzyme from the electrode and does not allow easy access of mediators to the surface. Attachment of the enzyme at smaller distance from the electrode by means of significantly shorter spacer molecule—mercaptopropionic acid improved the efficiency of catalytic reduction of oxygen on the monolayer modified electrode.