Enzymatic Fuel Cell Research Articles

Electrochemical Impedance Spectroscopy as a Characterization Method for Enzymatic Fuel Cell Bioanodes

Electrochemical Impedance Spectroscopy as a Characterization Method for Enzymatic Fuel Cell Bioanodes

High voltage flexible glucose/O2 fully printed hydrogel-based enzymatic fuel cell

Herein we report on a novel enzymatic fuel cell (EFC) based on stencil printed electrodes modified with pyrrolo quinoline quinone glucose dehydrogenase and bilirubin oxidase, which are assembled by considering two different configurations: (i) normal assembling in liquid electrolyte and (ii) six EFCs connected in series, each one comprising both bioanode and biocathode, coupled through a hydrogel-based electrolyte in a stack-like mode similar to a Voltaic pile. After a deep electrodes characterization, they are assembled according to the first configuration obtaining an open circuit voltage (OCV) of 0.562 ± 0.002 V. Moreover, the EFC performance are substantially improved by using the second configuration (six EFCs connected in series) obtaining an OCV of 2.36 ± 0.22 V with a maximum power output of 22.9 ± 0.9 μW at a cell voltage of 1.95 V (operating in 10 mM D-glucose). This innovative approach represents a proof-of-concept towards the development of renewable power sources and could serve as a critical step in powering implantable bioelectronics, such as pacemakers.

Gold nanoparticles activate hydrogenase synthesis and improve heterotrophic growth of Ralstonia eutropha H16.

Ralstonia eutropha is a facultative chemolithoautotrophic aerobic bacterium that grows using organic substrates or H2 and CO2. Hydrogenases (Hyds) are synthesized under lithoautotrophic, or energy-limited heterotrophic conditions and are used in enzyme fuel cells (EFC) as anodic catalysts. The effects of chemically synthesized gold nanoparticles (Au-NPs) on R. eutropha H16 growth, oxidation-reduction potential (ORP) kinetics, and H2-oxidizing Hyd activity were investigated in this study. Atomic force microscopy showed that thin, plate-shaped Au-NPs were in the nanoscale range with an average size of 5.68 nm. Compared with growth in medium without Au-NPs (control), the presence of Au-NPs stimulated growth, and resulted in a decrease in ORP to negative values. H2-oxidizing activity was not detected in the absence of Au-NPs, but activity was significantly induced (12 U/g CDW) after 24 h of growth with 18 ng/ml, increasing a further 4-fold after 72 h of growth. The results demonstrate that Au-NPs primarily influence the membrane-bound Hyd. In contrast to R. eutropha, Au-NPs had a negligible or negative effect on the growth, Hyd activity, and H2 production of Escherichia coli. The findings of this study offer new perspectives for the production of oxygen-tolerant Hyds and the development of EFCs.

Investigating Enzyme-Based Electrode Processes Using Electrochemical Impedance Spectroscopy and Distribution of Relaxation Time Analysis

Enzymatic fuel cells (EFCs) are an emerging class of electrochemical devices that have the potential to harness renewable energy sources by converting the chemical energy of an organic substrate into electrical energy. Enzymes are responsible in an EFC for highly selective and efficient electrode processes under typically mild ambient conditions, such as the oxidation reaction at the anode and the reduction reaction at the cathode 1. In addition to more conventional techniques, electrochemical impedance spectroscopy (EIS) can be employed to characterize such systems. Electrochemical behavior of enzymatic electrodes, such as enzyme reaction kinetics, substrate mass transport, and electrode stability, can be studied by measuring the system's impedance over a wide frequency range and then performing a precise post-process analysis based on the distribution of relaxation time (DRT). DRT analysis is rarely used to interpret EIS spectra within the EFCs research field, despite the fact that it is widely acknowledged as relevant for properly understanding the EIS potential to improve power density in fuel cells 2,3. Glucose oxidase (GOx) and bilirubin oxidase (BOD) were immobilized on multi-walled carbon nanotubes (MWCNTs) to catalyze the bioelectrocatalytic oxidation of glucose at the anode and the bioelectrocatalytic reduction of oxygen at the cathode, respectively. This study presents a promising approach for the investigation of enzyme-based anodic and cathodic processes by measuring them with EIS and then interpreting the results with DRT analysis, as depicted in Figure 1. By varying different operating parameters, such as the enzymatic loading on the electrode surface or electrolyte conditions, this method can be used to identify three primary relaxation processes occurring at both electrodes. At the interface between the electrolyte and the electrode, it is possible to conclude that high-frequency processes are accountable for ionic conduction, intermediate-frequency processes are accountable for charge transfer, and low-frequency processes are accountable for oxygen surface exchange and diffusion.This method provides access to preliminary fundamental information about enzymatic bioelectrode processes, which can be used to optimize biofuel cell construction and compositions and increase their power output. Figure 1. Schematic representation of the characterization method used to characterize a GOx-CNT-modified anode consisting of EIS measurements and subsequent DRT analysis. Concentration of glucose 175 mM in 0.1 M HEPES Buffer, pH 7; EIS parameters: 20 mV sinusoidal excitation, 0.1 Hz-100 kHz. References K. Herkendell, A. Stemmer and R. Tel-Vered, Nano Res., 12(4), 767–775 (2019). H. Wang, X. Long, Y. Sun, D. Wang, Z. Wang, H. Meng, C. Jiang, W. Dong and N. Lu, Frontiers in microbiology, 13, 973501 (2022). F. Ciucci, Current Opinion in Electrochemistry, 13, 132–139 (2019). Figure 1

Development of an enhanced hybrid bi-catalytic electrode containing bimetallic composite catalyst and immobilized enzyme for complete glucose electrooxidation

We describe an enzymatic fuel cell (EFC) that can electrooxidize glucose completely. The EFC contains the hybrid Ni@Pt-CNT/OxOx bioanode, composed of a bimetallic composite catalyst (Ni@Pt-CNT) and the enzyme oxalate oxidase (OxOx), which can cleave carbon-carbon bonds. Ni@Pt-CNT/OxOx displayed 3-fold higher catalytic activity in the presence than in the absence of glucose (1.3 and 0.4 mA cm−2, respectively), indicating that Ni@Pt-CNT and OxOx acted synergistically. Electrochemical impedance spectroscopy showed that Ni@Pt-CNT/OxOx had higher charge transfer resistance and double layer capacitance than Ni@Pt-CNT. Long-term bulk electrolysis (18 h) revealed that the EFC operating with Ni@Pt-CNT/OxOx presented better current density and stability than the electrochemical cell operating with Ni@Pt-CNT, so deep glucose electrooxidation generated energy. The glucose oxidation products detected by HPLC-UV/RID confirmed that glucose was fully electrooxidized, and that 24 electrons were harvested from it. The hybrid Ni@Pt-CNT/OxOx bioanode developed herein could be used in cascade reactions, to provide an EFC with promising application in self-powered electronic devices.

L-amino acids affect the hydrogenase activity and growth of Ralstonia eutropha H16

Ralstonia eutropha H16 is a chemolithoautotrophic bacterium with O2-tolerant hydrogenase (Hyds) enzymes. Hyds are expressed in the presence of gas mixtures (H2, O2, CO2) or under energy limitation and stress conditions. O2-tolerant Hyds are promising candidates as anode biocatalysts in enzymatic fuel cells (EFCs). Supplementation of 0.5% (w/v) yeast extract to the fructose-nitrogen (FN) growth medium enhanced H2-oxidizing Hyd activity ~ sixfold. Our study aimed to identify key metabolites (l-amino acids (l-AAs) and vitamins) in yeast extract that are necessary for the increased synthesis and activity of Hyds. A decrease in pH and a reduction in ORP (from + 240 ± 5 mV to − 180 mV ± 10 mV values) after 24 h of growth in the presence of AAs were observed. Compared to the FN-medium control, supplementation of 7.0 μmol/ml of the l-AA mixture stimulated the growth of bacteria ~ 1.9 to 2.9 fold, after 72 h. The whole cells’ H2-oxidizing Hyd activity was not observed in control samples, whereas the addition of l-AAs, mainly glycine resulted in a maximum of ~ 22 ± 0.5 and 15 ± 0.3 U, g CDW−1 activity after 24 h and 72 h, respectively. Our results suggest a correlation between ORP, pH, and function of Hyds in R. eutropha H16 in the presence of key l-AAs. l-AAs used in small amounts can be proposed as signaling molecules or key components of Hyd maturation. These results are important for the optimization of O2-tolerant Hyds production as anode biocatalysts.

Assembly of an improved hybrid cascade system for complete ethylene glycol oxidation: Enhanced catalytic performance for an enzymatic biofuel cell

We report an Enzymatic Fuel Cell (EFC) combining an enzyme that can cleave carbon-carbon bonds (oxalate oxidase (OxOx)) with an organic catalyst (Pyrene-TEMPO (TEMPO = 2,2,6,6-tetramethyl piperidinyl-N-oxyl)) immobilized on the surface of modified carboxylated multi-walled carbon nanotubes (MWCNT-COOH). This combination gave a hybrid bi-catalyst electrode for complete ethylene glycol (EG) oxidation. The hybrid electrode provided ninefold enhanced catalytic activity (0.17 ± 6 × 10−3 mA cm−2) in the presence of EG as compared to the electrode in the absence of EG (0.018 ± 3 × 10−5 mA cm−2), indicating that the enzyme combined with the organic catalyst improved energy generation through deep EG electrooxidation. Electrochemical impedance spectroscopy reveals that the addition of the enzyme in the electrode containing MWCNT–COOH–Pyrene-TEMPO increased the charge transfer resistance (Rct) and the capacitance of the double layer. Long-term electrolysis for 15 h showed that the hybrid electrode presented outstanding current density and stability. The EG oxidation products were identified and quantified by high-performance liquid chromatography (HPLC-UV/RID). The results confirmed complete EG oxidation in the presence of CO2 in the solution, allowing 10 electrons to be collected from the fuel. Overall, this study illustrates the development of a simple and improved hybrid bi-catalyst electrode for promising applications in small electronic devices.

Recent advancements in the field of flexible/wearable enzyme fuel cells

This review article focusses on new advances in the field of enzyme fuel cells (EFCs), especially, on flexible materials which can be used to make flexible EFCs for wearable devices, three-dimensional (3D) printed structures to prepare electrodes for EFCs and micro/nano electromechanical structures (MEMS/NEMS) to fabricate micro-EFCs. Particular attention is given to newly developed approaches to obtain efficient electrodes for harvesting energy via EFCs. This review article explains the various attributes of these recently developing technologies and their ability to mitigate the energy requirements of flexible/wearable bioelectronic devices. Besides discussing key milestones achieved, this perspective review article also emphasizes the main hurdles that are currently impeding the realization of flexible/wearable EFCs. We have also emphasized on the major hurdles related to the flexible materials required to fabricate wearable EFCs, suitable 3D printing techniques required, and MEMS and NEMS to fabricate micro-EFCs. In all the recently developed techniques, there are some common issues like stability, low power output, mechanical strength and flexibility. This review article also provides various routes to mitigate these issues. The main aim of this perspective article is to develop curiosity among the researchers of various fields to team up in order to address the huge challenges that restrict the real-world application of flexible/wearable EFCs. Such collaboration is important to harness the full potential of EFCs. It is requested to read this review article with supporting information to get the complete essence.

Sustainable and renewable electric energy generation with continuous flow enzymatic fuel cell

AbstractA continuous flow enzymatic fuel cell (EFC) based on poly(pyrrole‐co‐pyrrole‐2‐carboxylic acid) film was used for sustainable energy generation. The system parameters were optimized to generate maximum power. Optimum cycle number for electropolymerization was found to be 12.5 for bioanode, and 20 for biocathode. The best performance was reached at pH 7, 1 ml min−1 flow rate and cell potential of 0.51 V. A power density of 0.36 μW cm−2 in 10 min was generated with 0.86 μg glucose dehydrogenase and 0.35 μg bilirubin oxidase enzyme quantity by using 100 mmol L−1 glucose. The EFC performance was investigated with municipal sewage, and the system generated a power density of 0.85 μW cm−2. Results showed that the proposed EFC could be successfully used at waste utilization, especially for the wastewater with glycolytic content.

A Short Overview of Biological Fuel Cells.

This short review summarizes the improvements on biological fuel cells (BioFCs) with or without ionomer separation membrane. After a general introduction about the main challenges of modern energy management, BioFCs are presented including microbial fuel cells (MFCs) and enzymatic fuel cells (EFCs). The benefits of BioFCs include the capability to derive energy from waste-water and organic matter, the possibility to use bacteria or enzymes to replace expensive catalysts such as platinum, the high selectivity of the electrode reactions that allow working with less complicated systems, without the need for high purification, and the lower environmental impact. In comparison with classical FCs and given their lower electrochemical performances, BioFCs have, up to now, only found niche applications with low power needs, but they could become a green solution in the perspective of sustainable development and the circular economy. Ion exchange membranes for utilization in BioFCs are discussed in the final section of the review: they include perfluorinated proton exchange membranes but also aromatic polymers grafted with proton or anion exchange groups.

Improving the Performance of Baker’s Yeast-powered MFCs by Adding Dehydrogenase Enzymes to Oxidize Yeast-produced Ethanol

This study focuses on theoretically and experimentally enhancing the performance of a baker’s yeast-powered microbial fuel cell (MFC) by hybridizing the MFC and EFC (enzymatic fuel cells) technologies. To improve the power density of the MFC, commercially available alcohol and aldehyde dehydrogenase enzymes (ADHE) have been added to the anode chamber to oxidize ethanol, which is produced by Baker’s yeast (BKY) during the metabolic process, to acetic acid. This oxidation process contributes more power output for the MFC. BKY biofilm was formed in porous 3D activated carbon paper to make the anode. The combination of using the biofilm anode and ADHE for electricity generation by the MFC enabled about 19% improvement in the maximum power density if compared with the MFC without using ADHE. Also, aerobic and anaerobic culture conditions were investigated. The result showed that under the anaerobic culture condition, the MFC generated about 22% higher maximum power density than that of the aerobic condition. These results imply that the addition of ADHE can take full advantage of BKY-produced ethanol to boost overall BKY-powered MFC performance.

In Silico Analysis of Glucose Oxidase H516r and H516d Mutations for an Enzymatic Fuel Cell

Glucose oxidase (GOx) is an oxido-reductase enzyme that catalyzes glucose into hydrogen peroxide and glucono delta-lactone (GDL). GOx has the potential to be used in the medical field. Numerous research concerning the usage of GOx to create enzymatic biofuel cells have been done, nevertheless the results obtained have not been optimal. This research aims to increase the Km values of GOx in order to increase its potential as a material for an enzymatic fuel cell. The amino acid histidine in position 516 is a residue in the active site that plays an important part in the process of glucose oxidation. In this research we mutated H516 by in silico twice resulting in the mutants R516 and D516. The mutations resulted in a change of the docking area for both mutants and in the docking affinity for H516D resulting in higher Km values. This shows that the H516 residue plays an important part in the functions of glucose oxidase and mutation into aspartate could improve glucose oxidase based enzymatic fuel cells.

Separate immobilization of glucose oxidase and trehalase, and optimization of enzyme-carbon nanotube layers for the anode of enzymatic fuel cells utilizing trehalose

Insect cyborgs can be used as search robots in disaster areas and as environmental monitoring robots. In this study, we aimed to develop an anode for compact enzymatic fuel cells (EFCs) utilizing trehalose to power electronic devices implanted in such insects. Conventional trehalose-EFCs are fabricated by co-immobilization of glucose oxidase (GOx) and trehalase (TREH) on the surface of the anode. Here, we showed that current generation can be enhanced by separate immobilization of GOx and TREH using agarose. The effects of GOx-redox mediator layers, TREH loading, and the incorporation of single-walled carbon nanotubes (SWCNTs) on the performance of GOx-TREH anodes were investigated. The optimal arrangement of GOx-redox mediator layers with or without SWCNTs was determined. The highest oxidation current density (494 μA/cm2) was obtained at an electrode composed of three GOx-redox mediator layers and three GOx-redox mediator-SWCNTs layers, and agarose containing TREH. The bilirubin oxidase (BOD)-redox mediator layers and SWCNTs also influenced the performance of cathode. The highest reduction current intensity was obtained for the cathode with six BOD-redox mediator-SWCNTs layers. EFCs with the optimized anode and cathode showed a power density of ca. 15 μW/cm2 at a cell voltage of 0.3 V under a discharge current density of 50 μA/cm2.

Paper-based mediatorless enzymatic microfluidic biofuel cells

In this study, eco-friendly and disposable paper-based membraneless microfluidic enzymatic fuel cells (EFCs) were developed without any mediators to reduce the toxicity and cost of EFCs. Glucose oxidase and laccase were immobilized on multi-walled carbon nanotube electrodes to catalyze the redox reaction of glucose and oxygen. Micromachining techniques well-suited for mass production were used to precisely fabricate micro-scale Y-shaped and cross-shaped EFCs. Experimental measurements showed that the concentration of glucose in the fuel solution affects the cell performance, which occurs because the flow speed of the fuel stream decreases as the concentration of glucose increases. The highest performance of power density (104.2 ± 3.35 μW cm−2) and current density (615.6 ± 3.14 μA cm−2) were obtained with the Y-shaped channel configuration at a glucose concentration of 100 mM. This performance is the best of all paper-based single EFCs reported to date. The new paper-based co-laminar flow mediatorless EFC exhibits strong potential to power miniaturized and portable on-site diagnostic devices.

Bioelectrocatalytic Conversion of N2: From Chemically Inert Gas to Chiral Chemicals

Dinitrogen molecule (N2) is the most abundant natural gas and also the ultimate source of nitrogen for nitrogenated industrial and natural compounds. So far, the fully hydrogenated product, ammonia (NH3), is the most common product of N2 fixation.1 However, NH3, the regular end-product of N2 fixation, is a bulk chemical with low added-value. The conversion of NH3 to nitrogenous compounds with high added-value and complicated structure still requires further tedious chemical synthesis with the use of precious metal catalysts.2 In our study, an upgraded N2 fixation system was firstly designed and constructed based on a bioelectrocatalytic N2 reduction architecture.3 The NH3 generated via N2 reduction catalyzed by nitrogenase was in situ utilized by L-alanine dehydrogenase to realize the reductive amination of pyruvate and the generation of alanine. The reduced equivalent to support the N2 reduction and reductive amination of pyruvate was regenerated by the electrochemical method. ω-TA transferred the amino group from alanine to ketone substrates and finally produced desired chiral amine intermediates.4 On this basis, an H2 fuel cell module was integrated into the upgraded N2 fixation system to construct a self-powered H2/α-keto acid fuel cell. In the new system, the upgraded N2 fixation was powered by the oxidation of H2 with high Faradaic efficiency and without the requirement of external electrical energy input.5 Finally, the end-product of N2 fixation in our study successfully went beyond NH3 and reached high-value-added chiral amine intermediates and chiral amino acids. The produced chiral amine intermediates and chiral amino acids have wild application in pharmaceuticals production and biotechnology research. References (1) Foster, S. L.; Bakovic, S. I. P.; Duda, R. D.; Maheshwari, S.; Milton, R. D.; Minteer, S. D.; Janik, M. J.; Renner, J. N.; Greenlee, L. F. Catalysts for nitrogen reduction to ammonia. Nat. Catal. 2018, 1, 490.(2) Balaraman, E.; Srimani, D.; Diskin-Posner, Y.; Milstein, D. Direct synthesis of secondary amines from alcohols and ammonia catalyzed by a ruthenium pincer complex. Catal. Lett. 2015, 145, 139-144.(3) Milton, R. D.; Cai, R.; Abdellaoui, S.; Leech, D.; De Lacey, A. L.; Pita, M.; Minteer, S. D. Bioelectrochemical Haber-Bosch Process: An Ammonia-Producing H2/N2 Fuel Cell. Angew. Chem. Int. Ed. 2017, 56, 2680-2683.(4) Chen, H.; Cai, R.; Patel, J.; Dong, F.; Chen, H.; Minteer, S. D. Upgraded Bioelectrocatalytic N2 Fixation: From N2 to Chiral Amine Intermediates. J. Am. Chem. Soc. 2019, 141, 4963-4971.(5) Chen, H.; Prater, M. B.; Cai, R.; Dong, F.; Chen, H.; Minteer, S. D., Bioelectrocatalytic Conversion from N2 to Chiral Amino Acids in a H2/α-Keto Acid Enzymatic Fuel Cell. J. Am. Chem. Soc. 2020, 142 (8), 4028-4036.

Self-Powered Detection of Glucose by Enzymatic Glucose/Oxygen Fuel Cells on Printed Circuit Boards.

Monitoring glucose levels in physiological fluids can help prevent severe complications associated with hypo- and hyper-glycemic events. Current glucose-monitoring systems require a three-electrode setup and a power source to function, which can hamper the system miniaturization to the patient discomfort. Enzymatic fuel cells (EFCs) offer the opportunity to develop self-powered and minimally invasive glucose sensors by eliminating the need for an external power source. Nevertheless, practical applications demand for cost-effective and mass-manufacturable EFCs compatible with integration strategies. In this study, we explore for the first time the use of gold electrodes on a printed circuit board (PCB) for the development of an EFC and demonstrate its application in saliva. To increase the specific surface area, the PCB gold-plated electrodes were modified with porous gold films. At the anode, glucose oxidase is immobilized with an osmium redox polymer that serves as an electron-transfer mediator. At the cathode, bilirubin oxidase is adsorbed onto the porous gold surface with a blocking agent that prevents parasitic reactions while maintaining the enzyme catalytic activity. The resulting EFC showed a linear response to glucose in phosphate buffer within the range 50 μM to 1 mM, with a sensitivity of 14.13 μA cm–2 mM–1. The sensor was further characterized in saliva, showing the linear range of detection of 0.75 to 2 mM, which is within the physiological range, and sensitivity of 21.5 μA cm–2 mM–1. Overall, this work demonstrates that PCBs are suitable platforms for EFCs, paving the way for the development of fully integrated systems in a seamless and miniaturized device.

A Hybrid Microbial–Enzymatic Fuel Cell Cathode Overcomes Enzyme Inactivation Limits in Biological Fuel Cells

The construction of optimized biological fuel cells requires a cathode which combines the longevity of a microbial catalyst with the current density of an enzymatic catalyst. Laccase-secreting fungi were grown directly on the cathode of a biological fuel cell to facilitate the exchange of inactive enzymes with active enzymes, with the goal of extending the lifetime of laccase cathodes. Directly incorporating the laccase-producing fungus at the cathode extends the operational lifetime of laccase cathodes while eliminating the need for frequent replenishment of the electrolyte. The hybrid microbial–enzymatic cathode addresses the issue of enzyme inactivation by using the natural ability of fungi to exchange inactive laccases at the cathode with active laccases. Finally, enzyme adsorption was increased through the use of a functionally graded coating containing an optimized ratio of titanium dioxide nanoparticles and single-walled carbon nanotubes. The hybrid microbial–enzymatic fuel cell combines the higher current density of enzymatic fuel cells with the longevity of microbial fuel cells, and demonstrates the feasibility of a self-regenerating fuel cell in which inactive laccases are continuously exchanged with active laccases.

Recent Advances in the Direct Electron Transfer-Enabled Enzymatic Fuel Cells.

Direct electron transfer (DET), which requires no mediator to shuttle electrons from enzyme active site to the electrode surface, minimizes complexity caused by the mediator and can further enable miniaturization for biocompatible and implantable devices. However, because the redox cofactors are typically deeply embedded in the protein matrix of the enzymes, electrons generated from oxidation reaction cannot easily transfer to the electrode surface. In this review, methods to improve the DET rate for enhancement of enzymatic fuel cell performances are summarized, with a focus on the more recent works (past 10 years). Finally, progress on the application of DET-enabled EFC to some biomedical and implantable devices are reported.

Stability of Proton Exchange Membranes in Phosphate Buffer for Enzymatic Fuel Cell Application: Hydration, Conductivity and Mechanical Properties.

Proton-conducting ionomers are widespread materials for application in electrochemical energy storage devices. However, their properties depend strongly on operating conditions. In bio-fuel cells with a separator membrane, the swelling behavior as well as the conductivity need to be optimized with regard to the use of buffer solutions for the stability of the enzyme catalyst. This work presents a study of the hydrolytic stability, conductivity and mechanical behavior of different proton exchange membranes based on sulfonated poly(ether ether ketone) (SPEEK) and sulfonated poly(phenyl sulfone) (SPPSU) ionomers in phosphate buffer solution. The results show that the membrane stability can be adapted by changing the casting solvent (DMSO, water or ethanol) and procedures, including a crosslinking heat treatment, or by blending the two ionomers. A comparison with NafionTM shows the different behavior of this ionomer versus SPEEK membranes.

A glucose/oxygen enzymatic fuel cell exceeding 1.5 V based on glucose dehydrogenase immobilized onto polyMethylene blue-carbon nanotubes modified double-sided screen printed electrodes: Proof-of-concept in human serum and saliva

Herein we report on a new hybrid enzymatic fuel cell (EFC) based on mediated/direct electron transfer, which is assembled by considering two different configurations: i) two normal screen printed electrodes (SPEs) in one electrochemical cell and ii) two double-sided SPEs each one comprising both bioanode and biocathode, connected in series.After a deep electrodes characterization, they are assembled according to the first configuration obtaining a maximal power output of 25.8 ± 0.7 μW cm−2 at a cell voltage of 0.57 V and an open circuit voltage (OCV) of 0.78 V. Moreover, the EFC performance are substantially improved by using the second configuration (EFCs connected in series) obtaining a maximal power output of 59.6 ± 1.9 μW cm−2 at a cell voltage of 1.16 V and an open circuit voltage (OCV) of 1.58 V. Finally, the last EFC configuration is tested in human saliva and serum showing good performance in terms of OCV and maximal power output as proof-of-concept to power implantable bioelectronics (e.g., pacemaker) or develop self-powered glucose biosensors.