Reagentless D-Tagatose Biosensors Based on the Oriented Immobilization of Fructose Dehydrogenase onto Coated Gold Nanoparticles- or Reduced Graphene Oxide-Modified Surfaces: Application in a Prototype Bioreactor

As electrode nanomaterials, thermally reduced graphene oxide (TRGO) and modified gold nanoparticles (AuNPs) were used to design bioelectrocatalytic systems for reliable D-tagatose monitoring in a long-acting bioreactor where the valuable sweetener D-tagatose was enzymatically produced from a dairy by-product D-galactose. For this goal D-fructose dehydrogenase (FDH) from Gluconobacter industrius immobilized on these electrode nanomaterials by forming three amperometric biosensors: AuNPs coated with 4-mercaptobenzoic acid (AuNP/4-MBA/FDH) or AuNPs coated with 4-aminothiophenol (AuNP/PATP/FDH) monolayer, and a layer of TRGO on graphite (TRGO/FDH) were created. The immobilized FDH due to changes in conformation and spatial orientation onto proposed electrode surfaces catalyzes a direct D-tagatose oxidation reaction. The highest sensitivity for D-tagatose of 0.03 ± 0.002 μA mM–1cm–2 was achieved using TRGO/FDH. The TRGO/FDH was applied in a prototype bioreactor for the quantitative evaluation of bioconversion of D-galactose into D-tagatose by L-arabinose isomerase. The correlation coefficient between two independent analyses of the bioconversion mixture: spectrophotometric and by the biosensor was 0.9974. The investigation of selectivity showed that the biosensor was not active towards D-galactose as a substrate. Operational stability of the biosensor indicated that detection of D-tagatose could be performed during six hours without loss of sensitivity.

Carbon-Encapsulated Iron Nanoparticles as a Magnetic Modifier of Bioanode and Biocathode in a Biofuel Cell and Biobattery

This work demonstrates the application of magnetic carbon-encapsulated iron nanoparticles (CEINs) for the construction of bioelectrodes in a biobattery and a biofuel cell. It has been shown that carbon-encapsulated iron nanoparticles are a suitable material for the immobilization of laccase (Lc) and 1,4-naphthoquinone (NQ) and fructose dehydrogenase (FDH). The system is stable; no leaching of the enzyme and mediator from the surface of the modified electrode was observed. The onset of the catalytic reduction of oxygen to water was at 0.55 V, and catalytic fructose oxidation started at −0.15 V. A biobattery was developed in which a zinc plate served as the anode, and the cathode was a glassy carbon electrode modified with carbon-encapsulated iron nanoparticles, laccase in the Nafion (Nf) layer. The maximum power of the cell was ca. 7 mW/cm2 at 0.71 V and under external resistance of 1 kΩ. The open-circuit voltage (OCV) for this system was 1.51 V. In the biofuel cell, magnetic nanoparticles were used both on the bioanode and biocathode to immobilize the enzymes. The glassy carbon bioanode was coated with carbon-encapsulated iron nanoparticles, 1,4-naphthoquinone, fructose dehydrogenase, and Nafion. The cathode was modified with carbon-encapsulated magnetic nanoparticles and laccase in the Nafion layer. The biofuel cell parameters were as follows: maximum power of 78 µW/cm2 at the voltage of 0.33 V and under 20 kΩ resistance, and the open-circuit voltage was 0.49 V. These enzymes worked effectively in the biofuel cell, and laccase also effectively worked in the biobattery.