Biosensor development for low-level acetaldehyde gas detection using mesoporous carbon electrode printed on a porous polyimide film.

Acetaldehyde, which is an intermediate product of alcohol metabolism, is known to induce symptoms, including alcohol flushing, vomiting, and headaches in humans. Therefore, real-time monitoring of acetaldehyde levels is crucial to mitigating these health issues. However, current methods for detecting low-concentration gases necessitate the use of complex measurement equipment. In this study, we developed a low-cost, low-detection-limit, enzyme-based electrochemical biosensor for acetaldehyde gas detection that does not require sophisticated equipment. The sensor was constructed by screen-printing electrodes onto a porous polyimide film, using grafted MgO-templated carbon (GMgOC) as working electrode material, carbon for the counter electrode, and silver/silver chloride for the reference electrode. Pyrroloquinoline-quinone-dependent aldehyde dehydrogenase was immobilized on the working electrode, and a chamber was attached to the electrode chip and filled with 1-methoxy-5-methylphenazinium methyl sulfate solution. The sensor can be used to measure acetaldehyde gas concentrations from 0.02 to 0.1 ppm, making it suitable for monitoring human skin gas. This low detection limit was achieved by delivering the analyte through the porous polyimide film on which the electrodes were printed and accumulating acetaldehyde in the mesoporous GMgOC of the working electrode. This mechanism suggests that this sensor design can be adapted to develop other low-detection limit gas sensors, such as those for screening skin gas biomarkers.

Wearable Ion Sensors for the Detection of Sweat Ions Fabricated by Heat-Transfer Printing.

Wearable ion sensors for the real-time monitoring of sweat biomarkers have recently attracted increasing research attention. Here, we fabricated a novel chloride ion sensor for real-time sweat monitoring. The printed sensor was heat-transferred onto nonwoven cloth, allowing for easy attachment to various types of clothing, including simple garments. Additionally, the cloth prevents contact between the skin and the sensor and acts as a flow path. The change in the electromotive force of the chloride ion sensor was -59.5 mTV/log CCl-. In addition, the sensor showed a good linear relationship with the concentration range of chloride ions in human sweat. Moreover, the sensor displayed a Nernst response, confirming no changes in the film composition due to heat transfer. Finally, the fabricated ion sensors were applied to the skin of a human volunteer subjected to an exercise test. In addition, a wireless transmitter was combined with the sensor to wirelessly monitor ions in sweat. The sensors showed significant responses to both sweat perspiration and exercise intensity. Thus, our research demonstrates the potential of using wearable ion sensors for the real-time monitoring of sweat biomarkers, which could significantly impact the development of personalized healthcare.

Air-Bubble-Insensitive Microfluidic Lactate Biosensor for Continuous Monitoring of Lactate in Sweat.

This study aimed to develop a lactate sensor with a microchannel that overcomes the issue of air bubbles interfering with the measurement of lactate levels in sweat and to evaluate its potential for continuous monitoring of lactate in sweat. To achieve continuous monitoring of lactate, a microchannel was used to supply and drain sweat from the electrodes of the lactate sensor. A lactate sensor was then developed with a microchannel that has an area specifically designed to trap air bubbles and prevent them from contacting the electrode. The sensor was evaluated by a person while exercising to test its effectiveness in monitoring lactate in sweat and its correlation with blood lactate levels. Furthermore, the lactate sensor with a microchannel in this study can be worn on the body for a long time and is expected to be used for the continuous monitoring of lactate in sweat. The developed lactate sensor with a microchannel effectively prevented air bubbles from interfering with the measurement of lactate levels in sweat. The sensor showed a concentration correlation ranging from 1 to 50 mM and demonstrated a correlation between lactate in sweat and blood. Additionally, the lactate sensor with a microchannel in this study can be worn on the body for an extended period and is expected to be useful for the continuous monitoring of lactate in sweat, particularly in the fields of medicine and sports.

High-performance paper-based biocathode fabricated by screen-printing an improved mesoporous carbon ink and by oriented immobilization of bilirubin oxidase.

In this study, the performance of a paper-based, screen-printed biofuel cell with mesoporous MgO-templated carbon (MgOC) electrodes was improved in two steps. First, a small amount of carboxymethyl cellulose (CMC) was added to the MgOC ink. Next, the cathode was modified with bilirubin prior to immobilizing the bilirubin oxidase (BOD). The CMC increased the accessibility of the mesopores of the MgOC, and subsequently, the performance of both the bioanode and biocathode. CMC also likely increased the stability of the electrodes. The pre-modification with bilirubin improved the orientation of the BOD, which facilitated direct electron transfer. With these two steps, an open circuit potential of 0.65 V, a maximal current density of 1.94 mA cm-2, and a maximal power density of 465 µW cm-2 was achieved with lactate oxidase as bioanode enzyme and lactate as fuel. This is one of the highest reported performances for a biofuel cell.

Self-Powered Diaper Sensor with Wireless Transmitter Powered by Paper-Based Biofuel Cell with Urine Glucose as Fuel.

A self-driven sensor that can detect urine and urine sugar and can be mounted on diapers is desirable to reduce the burden of long-term care. In this study, we created a paper-based glucose biofuel cell that can be mounted on diapers to detect urine sugar. Electrodes for biofuel cells were produced by printing MgO-templated porous carbon on which poly(glycidyl methacrylate) was modified using graft polymerization. A new bioanode was prepared through covalently modifying flavin-adenine-dinucleotide-dependent glucose dehydrogenase and azure A with pendant glycidyl groups of poly(glycidyl methacrylate). We prepared a cathode with covalently bonded bilirubin oxidase. Covalent bonding of enzymes and mediators to both the bioanode and biocathode suppressed elution and improved stability. The biofuel cell could achieve a maximum output density of 0.12 mW cm-2, and by combining it with a wireless transmission device, the concentration of glucose sensed from the transmission frequency was in the range of 0-10 mM. The sensitivity of the sensor was estimated at 0.0030 ± 0.0002 Hz mmol-1 dm3. This device is expected to be a new urine-sugar detection device, composed only of organic materials with a low environmental load and it can be useful for detecting postprandial hyperglycemia.

A screen-printed three-electrode-type sticker device with an accurate liquid junction-type reference electrode.

We developed a novel sticker device that can convert any metal or alloy into the working electrode of a three-electrode system, enabling simple and accurate measurement. The sticker, containing a counter electrode and a stable and accurate liquid junction-type reference electrode, is attached to the metal or alloy; meanwhile the surface exposed from a hole in the device functions as the working electrode. This sticker device was fabricated by screen-printing. The polarization curve of the copper and tin-plated copper measured using the sticker device exhibited approximately the same behavior as that obtained for the conventional three-electrode system. The characteristics of various materials can be easily evaluated using this system by only dropping a small amount of solution.

Analysis of porous carbon biocathodes via three-dimensional impedance spectroscopy using a double channel transmission line model.

Porous carbon electrodes have considerably improved the performance of biofuel cells and biosensors in recent years. In this paper, we propose a novel in-situ analysis method for porous enzyme electrodes. By combining three-dimensional (3D) impedance measurement and a double-channel transmission line model, the stability of porous enzyme electrodes during operation can be evaluated. The proposed method can distinguish between the functional stability of the enzyme and mediator reaction and the general structural stability of the electrode. We demonstrated this method by evaluating bilirubin oxidase-modified carbon cloth (CC) electrodes with and without a magnesium oxide (MgO)-templated carbon coating. In case of the CC electrode, a remarkable increase in the charge transfer resistance within the first 500 s indicated the elution of the enzyme and mediator. When the CC was coated with MgO-templated carbon before enzyme modification, the charge transfer resistance remained constant, indicating an effective suppression of the elution of the enzyme and mediator. The electric double-layer capacitance values of both electrodes indicated that their general electrode structures were stable during the analysis. Thus, the proposed analytical method, based on 3D impedance, can be a powerful tool for simultaneously detecting possible changes in the general electrode structure of enzyme electrodes and in the amount of active enzymes and mediators on the electrode surface.

Chitosan-based enzyme ink for screen-printed bioanodes.

In this study, magnesium oxide (MgO)-templated mesoporous carbon (MgOC) and chitosan cross-linked with genipin (chitosan-genipin) were considered bio-composite inks for screen-printed bioanodes. The fabrication processes were optimized using rheological and structural data, and a bioanode ink containing glucose oxidase (GOx) and 1,2-naphthoquinone (1,2-NQ) was successfully developed. The optimal bioanode-ink contained MgOC pre-treated by washing to achieve a hydrophilic and neutral surface, which helped maintain enzyme activity and resulted in a highly porous electrode structure, which is essential for the accessibility of glucose to GOx. A bioanode fabricated using this ink showed a linear response current dependency up to 8 mM glucose with a sensitivity of 25.83 µA cm-2 mM-1. Combined with a conventional biocathode, an electromotive force of 0.54 V and a maximal power density of 96 µW cm-2 were achieved. These results show that this bio-composite ink can be used to replace the multi-step process of printing with conventional ink followed by drop-casting enzyme and mediator with a one-step printing process.

Toward Wearable Energy Storage Devices: Paper-Based Biofuel Cells based on a Screen-Printing Array Structure.

A novel paper-based biofuel cell with a series/parallel array structure has been fabricated, in which the cell voltage and output power can easily be adjusted as required by printing. The output of the fabricated 4-series/4-parallel biofuel cell reached 0.97±0.02 mW at 1.4 V, which is the highest output power reported to date for a paper-based biofuel cell. This work contributes to the development of flexible, wearable energy storage device.

Enhanced Sensitivity for Electrochemical Detection Using Screen-Printed Diamond Electrodes via the Random Microelectrode Array Effect.

The electrochemical properties of screen-printed diamond electrodes with various insulating polyester (PES) resin binder/boron-doped diamond powder (BDDP) ratios were investigated for high sensitivity electrochemical detection. For PES/BDDP weight ratios in the range of 0.3-0.5, the BDDP-printed electrodes exhibited cyclic voltammetry (CV) characteristics for Fe(CN)6(3-/4-) that are typical of a planar electrode, whereas microelectrode-like characteristics with sigmoidal CV curves were observed for PES/BDDP ratios of 1.0-2.0. Cu elemental mapping images of copper-electrodeposited BDDP-printed electrodes indicated the formation of island structures with conductive BDDP domains surrounded by an insulating PES matrix for large PES/BDDP ratios. The electrochemical detection of ascorbic acid (AA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) was also investigated using polycrystalline BDD thin-film and BDDP-printed electrodes (PES/BDDP ratio = 0.3 and 1.0). As a result, the signal-to-background (S/B) ratios for the voltammetric detection of AA and 8-OHdG were in the order BDDP-printed electrode (PES/BDDP = 1.0) > BDDP-printed electrode (PES/BDDP = 0.3) > polycrystalline BDD thin film electrode, based on the large faradaic current with respect to the background current. Therefore, the BDDP-printed electrode with a large insulating binder/BDDP ratio has the potential for use as a disposable electrode for electrochemical detection. The electrode is cheaper, lighter and more sensitive than conventional BDD electrodes.

An instantly usable paper-based screen-printed solid-state KCl/Ag/AgCl reference electrode with long-term stability.

An instantly usable screen-printed paper-based Ag/AgCl electrode was fabricated for use as a cost-effective disposable reference electrode. The reference electrode showed potential stability for approximately 75 h. The setup time, which is less than 1 min, is much shorter than those for similar previously reported electrodes.

Electrochemical monitoring systems of demembranated flagellate algal motility for ATP sensing.

The ATP-induced behavior of the unicellular flagellate alga Chlamydomonas reinhardtii was recorded as changes in the redox currents for a coexisting redox marker. The ATP concentration was estimated using the presented compact electrochemical system, which is based on monitoring of the motility of the flagellates.

Flexible and high-performance paper-based biofuel cells using printed porous carbon electrodes.

We demonstrate a flexible paper-based biofuel cell using porous carbon inks for high power output. The power density of the fabricated biofuel cell reached 0.12 mW cm(-2) (at 0.4 V), which is the highest output power reported to date, to the best of our knowledge.

Elimination of influence by interference substance on amperometric enzyme biosensor response using wavelet transformation.

Wavelet transformation was applied as an elimination method of influence by an interfering substance on an amperometric biosensor. The current responses of a bi-enzyme type lactose biosensor in a solution containing both lactose and ascorbic acid were analyzed by wavelet transformation. The power spectrum density for ascorbic acid was detected selectively at around 0.125 Hz. The current component due to ascorbic acid was eliminated at 98% from the current response of the biosensor by wavelet transformation.

Wavelet transformation of amperometric algal biosensor response.

Wavelet transformation was applied as a noise elimination method of an amperometric algal biosensor. The drift of the baseline current was clearly removed by using the wavelet transformation. The S/N ratio, calculated by the power spectrum density, is about 3-times larger than that calculated by the current response. The response to a herbicide, atrazine, calculated from the power spectrum density in high-noise circumstance, was the same as that calculated from the current response in a low-noise circumstance.

Screen-printed dissolved oxygen sensor based on cerium oxide-supported silver catalyst and polydimethylsiloxane film.

A screen-printed dissolved oxygen sensor was fabricated using cerium oxide-supported silver catalyst and polydimethylsiloxane (PDMS) film. A PDMS film of 3 µm thickness showed good permeability for oxygen and impermeability for hydrogen peroxide. The calibration curve has shown a linear relationship with a correlation coefficient of 0.996 for the dissolved oxygen concentration. The sensitivity and detection limit of the present sensor were calculated at -158 µA mM(-1) and 8.4 µM, respectively.

Use of a Gluconobacter frateurii mutant to prevent dihydroxyacetone accumulation during glyceric acid production from glycerol.

To prevent dihydroxyacetone (DHA) by-production during glyceric acid (GA) production from glycerol using Gluconobacter frateurii, we used a G. frateurii THD32 mutant, ΔsldA, in which the glycerol dehydrogenase subunit-encoding gene (sldA) was disrupted, but ΔsldA grew much more slowly than the wild type, growth starting after a lag of 3 d under the same culture conditions. The addition of 1% w/v D-sorbitol to the medium improved both the growth and the GA productivity of the mutant, and ΔsldA produced 89.1 g/l GA during 4 d of incubation without DHA accumulation.

Two-stage electrodialytic concentration of glyceric acid from fermentation broth.

The aim of this research was the application of a two-stage electrodialysis (ED) method for glyceric acid (GA) recovery from fermentation broth. First, by desalting ED, glycerate solutions (counterpart is Na+) were concentrated using ion-exchange membranes, and the glycerate recovery and energy consumption became more efficient with increasing the initial glycerate concentration (30 to 130 g/l). Second, by water-splitting ED, the concentrated glycerate was electroconverted to GA using bipolar membranes. Using a culture broth of Acetobacter tropicalis containing 68.6 g/l of D-glycerate, a final D-GA concentration of 116 g/l was obtained following the two-stage ED process. The total energy consumption for the D-glycerate concentration and its electroconversion to D-GA was approximately 0.92 kWh per 1 kg of D-GA.

Microbial production of glyceric acid, an organic acid that can be mass produced from glycerol.

Glyceric acid (GA), an unfamiliar biotechnological product, is currently produced as a small by-product of dihydroxyacetone production from glycerol by Gluconobacter oxydans. We developed a method for the efficient biotechnological production of GA as a target compound for new surplus glycerol applications in the biodiesel and oleochemical industries. We investigated the ability of 162 acetic acid bacterial strains to produce GA from glycerol and found that the patterns of productivity and enantiomeric GA compositions obtained from several strains differed significantly. The growth parameters of two different strain types, Gluconobacter frateurii NBRC103465 and Acetobacter tropicalis NBRC16470, were optimized using a jar fermentor. G. frateurii accumulated 136.5 g/liter of GA with a 72% d-GA enantiomeric excess (ee) in the culture broth, whereas A. tropicalis produced 101.8 g/liter of d-GA with a 99% ee. The 136.5 g/liter of glycerate in the culture broth was concentrated to 236.5 g/liter by desalting electrodialysis during the 140-min operating time, and then, from 50 ml of the concentrated solution, 9.35 g of GA calcium salt was obtained by crystallization. Gene disruption analysis using G. oxydans IFO12528 revealed that the membrane-bound alcohol dehydrogenase (mADH)-encoding gene (adhA) is required for GA production, and purified mADH from G. oxydans IFO12528 catalyzed the oxidation of glycerol. These results strongly suggest that mADH is involved in GA production by acetic acid bacteria. We propose that GA is potentially mass producible from glycerol feedstock by a biotechnological process.

Production of glyceric acid by Gluconobacter sp. NBRC3259 using raw glycerol.

Gluconobacter sp. NBRC3259 converted glycerol to glyceric acid (GA). The enantiomeric composition of the GA produced was a mixture of DL-forms with a 77% enantiomeric excess of D-GA. After culture conditions, such as initial glycerol concentration, types and amounts of nitrogen sources, and initial pH, were optimized, Gluconobacter sp. NBRC3259 produced 54.7 g/l of GA as well as 33.7 g/l of dihydroxyacetone (DHA) from 167 g/l of glycerol during 4 d of incubation in a jar fermentor with pH control. GA production from raw glycerol samples, the main by-product of the transesterification process in the biodiesel production and oleochemical industries, was also evaluated after proper pretreatment of the samples. Using a raw glycerol sample with activated charcoal pretreatment, 45.9 g/l of GA and 28.2 g/l of DHA were produced from 174 g/l of glycerol.