Direct Electron Transfer between a Site-Specific Pyrene-Modified Laccase and Carbon Nanotube/Gold Nanoparticle Supramolecular Assemblies for Bioelectrocatalytic Dioxygen Reduction

Strategies to maximize direct electron transfer (DET) between redox enzymes and electrodes include the oriented immobilization of enzymes onto an electroactive surface. Here, we present a strategy for achieving a controlled orientation of a fungal laccase on carbon nanotube-based electrodes. A homogeneous population of pyrene-modified laccase is obtained via the reductive amination of a unique surface accessible lysine residue engineered near the T1 copper center of the enzyme. Immobilization of the site-specific functionalized enzyme is achieved either via pi-stacking of pyrene on pristine CNT electrodes or through pyrene/beta-cyclodextrin host guest interactions on beta-cyclodextrin-modified gold nanoparticles (beta-CD-AuNPs). Contrasting with unmodified and nonspecifically modified (pyrene-NHS) laccase-electrodes, an efficient DET is obtained at these nanostructured assemblies. Modeling the direct bioelectrocatalysis of dioxygen reduction reveals a heterogeneity in ET rates on MWCNT electrodes wheras beta-CD-AuNPs act as efficient electronic bridges, lowering ET rate dispersion and achieving a highly efficient reduction of O-2 at low overpotential (approximate to 80 mV) accompanied by high catalytic current densities of almost 3 mA cm(-2).