A lactate/O enzymatic biofuel cell (EBFC) was prepared as a potential power source for wearable microelectronic devices. Mechanically stable and flexible nanoporous gold (NPG) electrodes were prepared using an electrochemical dealloying method consisting of a pre-anodization process and a subsequent electrochemical cleaning step. Bioanodes were prepared by the electrodeposition of an Os polymer and Pediococcus sp. lactate oxidase onto the NPG electrode. The electrocatalytic response to lactate could be tuned by adjusting the deposition time. Bilirubin oxidase from Myrothecium verrucaria was covalently attached to a diazonium-modified NPG surface. A flexible EBFC was prepared by placing the electrodes between two commercially available contact lenses to avoid direct contact with the eye. When tested in air-equilibrated artificial tear solutions (3 mM lactate), a maximum power density of 1.7 ± 0.1 μW cm and an open-circuit voltage of 380 ± 28 mV were obtained, values slightly lower than those obtained in phosphate buffer solution (2.4 ± 0.2 μW cm and 455 ± 21 mV, respectively). The decrease was mainly attributed to interference from ascorbate. After 5.5 h of operation, the EBFC retained 20% of the initial power output.
Nanoporous gold (NPG) fabricated by sputtering is a material of versatile morphology with pores whose size can be tailored to accommodate enzymes. The process of pore formation and the size of the pores in NPG are influenced by the composition of Au and Ag in the alloy used to prepare the electrodes together with the temperature and time period of the dealloying process. On increasing the time from 1 to 60 min and the temperature from 0.5 °C to 60.5 °C in concentrated HNO3, significant increases in the average pore diameters from 4.4 to 78 nm were observed with simultaneous decreases in the roughness factor (Rf). The pores of NPG were fully addressable regardless of the diameter, with Rf increasing linearly up to an alloy thickness of 500 nm. The influence of the pore size on the bioelectrochemical response of redox proteins was evaluated using cytochrome c as a model system. The highest current densities of ca. 30 µA cm−2 were observed at cytochrome c modified NPG electrodes with an average pore size of ca. 10 nm. The pores in NPG were also tuned for the mediatorless immobilization of Myrothecium verrucaria bilirubin oxidase. High current densities of ca. 65 µA cm−2 were observed at MvBOD modified NPG electrodes prepared by dealloying at 0.5 °C for 5 min with an average pore size of 8 nm, which is too small to accommodate the enzyme into the pores, indicating that the response was from enzyme adsorbed on the electrode surface.
Nanoporous gold (NPG) electrodes were prepared by dealloying sputtered gold:silver alloys. Electrodes of different thicknesses and pore sizes areas were prepared by varying the temperature and duration of the dealloying procedure; these were then used as supports for FAD‐dependent glucose dehydrogenase (GDH) (Glomorella cingulata) and bilirubin oxidase (BOx) (Myrothecium verrucaria). Glucose dehydrogenase was immobilized by drop‐casting a solution of the enzyme with an osmium redox polymer together with a crosslinked polymer, whereas bilirubin oxidase was attached covalently through carbodiimide coupling to a diazonium‐modified NPG electrode. The stability of the bilirubin‐oxidase‐modified NPG electrode was significantly improved in comparison with that of a planar gold electrode. Enzyme fuel cells were also prepared; the optimal response was obtained with a BOx‐modified NPG cathode (500 nm thickness) and a GDH‐modified anode (300 nm), which generated power densities of 17.5 and 7.0 μW cm−2 in phosphate‐buffered saline and artificial serum, respectively.
Nanoporous gold electrodes were utilized as a support for the detection of d‐fructose. The immobilization of fructose dehydrogenase was achieved by adsorption on thiol‐ and diazonium‐bound carboxylic acid functional groups and subsequent crosslinking through cabodiimide‐initiated amide bond formation. The biosensor showed a linear response in the range 0.05–0.3 mM of d‐fructose with a sensitivity of 3.7±0.2 μA cm−2 mM−1 and a limit of detection of 1.2 μM. The response of the biosensor in a range of natural sweeteners and beverages compared very favorably to the results obtained by a commercially available kit. Accurate readings were obtainable after a very fast response time of less than 5 seconds. The biosensor showed high specificity towards d‐fructose in the presence of interfering sugars.
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