Real-time monitoring abscisic acid release from single rice protoplast by amperometry at microelectrodes modified with abscisic acid receptor PYL2.

It was previously reported that stress induces a cellular production of abscisic acid in plants, but no direct method shows the evidence. Here, an electrochemical microsensor involving an abscisic acid receptor PYL2 modified carbon fiber microelectrode was fabricated by self-assembly method, where the Cu2+ combined with the histidine tag of PYL2 on the surface of microelectrode was used as the detection probe, the mediated reaction between Cu+ and ferricyanide realized the amplification responses and provided the microsensor with a high sensitivity for detection of abscisic acid with a detection limit of 0.8 nM. With use of this microsensor, an increase of extracellular abscisic acid from single rice protoplast induced by sulfate, osmotic and salinity stress was real-time monitored. Direct measurement of free extracellular abscisic acid in single plant cells might offer important new insights into its role in plants challenged by abiotic stresses.

Silver oxide doped iron oxide/alginate nanocomposite coated cotton cloth for selective catalytic reduction of potassium ferricyanide.

Silver oxide doped iron oxide (Ag2O-Fe2O3) nanocatalyst was prepared and coated on cotton cloth (CC) as well as wrapped in sodium alginate (Alg) hydrogel. Ag2O-Fe2O3 coated CC (Ag2O-Fe2O3/CC) and Ag2O-Fe2O3 wrapped Alg (Ag2O-Fe2O3/Alg) were utilized as catalysts in reduction reaction of 4-nitrophenol (4-NP), congo red (CR), methylene blue (MB) and potassium ferricyanide (K3[Fe(CN)6]). Ag2O-Fe2O3/CC and Ag2O-Fe2O3/Alg were found to be effective and selective catalyst for the reaction of K3[Fe(CN)6]. Further amount of catalyst, K3[Fe(CN)6] quantity, amount of NaBH4, stability of catalyst and recyclability were optimized for the reaction of K3[Fe(CN)6] reduction. Ag2O-Fe2O3/Alg and Ag2O-Fe2O3/CC were appeared to be the stable catalysts by maintaining high activity during recyclability tests showing highest reaction rate constants (kapp) of 0.3472 and 0.5629 min-1, correspondingly. However, Ag2O-Fe2O3/CC can be easily recovered as compared to Ag2O-Fe2O3/Alg by simply removing from the reaction which is the main advantage of Ag2O-Fe2O3/CC. Moreover, Ag2O-Fe2O3/Alg and Ag2O-Fe2O3/CC were also examined in real samples and found useful for K3[Fe(CN)6] reduction involving real samples. The Ag2O-Fe2O3/CC nanocatalyst is a cost and time saving material for economical reduction of K3[Fe(CN)6] and environmental safety.

Chemoorganotrophic electrofermentation by Cupriavidus necator using redox mediators.

The non-pathogenic ß-proteobacterium Cupriavidus necator has the ability to switch between chemoorganotrophic, chemolithoautotrophic and electrotrophic growth modes, making this microorganism a widely used host for cellular bioprocesses. Oxygen usually acts as the terminal electron acceptor in all growth modes. However, several challenges are associated with aeration, such as foam formation, oxygen supply costs, and the formation of an explosive gas mixture in chemolithoautotrophic cultivation with H2, CO2 and O2. Bioelectrochemical systems in which O2 is replaced by an electrode as a terminal electron acceptor offer a promising solution to these problems. The aim of this study was to establish a mediated electron transfer between the anode and the metabolism of living cells, i.e. anodic respiration, using fructose as electron and carbon source. Since C. necator is not able to transfer electrons directly to an electrode, redox mediators are required for this process. Based on previous observations on the extracellular electron transfer enabled by a polymeric mediator, we tested 11 common biological and non-biological redox mediators for their functionality and inhibitory effect for anodic electron transfer in a C. necator-based bioelectrochemical system. The use of ferricyanide at a concentration of 15 mM resulted in the highest current density of 260.75µAcm-2 and a coulombic efficiency of 64.1 %.

Rapid and directly interpretable antimicrobial susceptibility profiling by continuous microvolume-electroanalysis of ferricyanide-mediated bacterial respiration.

We present a novel method for the electroanalysis of potassium ferricyanide-mediated bacterial electron transport, to rapidly assess viability and construct interpretable antimicrobial susceptibility profiles. Electrochemical minimum inhibitory concentrations (ecMICs) became determinable with a high correlation to the results from conventional assays.

Electrocatalytic aptasensor for bacterial detection exploiting ferricyanide reduction by methylene blue on mixed PEG/aptamer monolayers.

Pathogen-triggered infections are the most severe global threat to human health, and to provide their timely treatment and prevention, robust methods for rapid and reliable identification of pathogenic microorganisms are required. Here, we have developed a fast and inexpensive electrocatalytic aptamer assay enabling specific and ultrasensitive detection of E. coli. E. coli, a biomarker of environmental contamination and infections, was captured on the mixed aptamer/thiolated PEG self-assembled monolayers formed on electrochemically pre-treated gold screen-printed electrodes (SPE). Signals from aptamer - E. coli binding were amplified by electrocatalytic reduction of ferricyanide mediated by methylene blue (MB) adsorbed on bacterial and aptamer surfaces. PEG operated as an antifouling agent and inhibited direct (not MB-mediated) discharge of ferricyanide. The assay allowed from 10 to 1000 CFU mL-1E. coli detection in 30 min, with no interference from B. subtilis, in buffer and artificial urine samples. This electrocatalytic approach is fast, specific, sensitive, and can be used directly in in-field and point-of-care applications for analysis of bacteria in human environment.

Roadmap of electrons from donor side to the reaction center of photosynthetic purple bacteria with mutated cytochromes.

In photosynthetic bacteria, the absorbed light drives the canonical cyclic electron transfer between the reaction center and the cytochrome bc1 complexes via the pools of mobile electron carriers. If kinetic or structural barriers hinder the participation of the bc1 complex in the cyclic flow of electrons, then the pools of mobile redox agents must supply the electrons for the multiple turnovers of the reaction center. These conditions were achieved by continuous high light excitation of intact cells of bacterial strains Rba. sphaeroides and Rvx. gelatinosus with depleted donor side cytochromes c2 (cycA) and tetraheme cytochrome subunit (pufC), respectively. The gradual oxidation by ferricyanide further reduced the availability of electron donors to pufC. Electron transfer through the reaction center was tracked by absorption change and by induction and relaxation of the fluorescence of the bacteriochlorophyll dimer. The rate constants of the electron transfer (~ 3 × 103 s‒1) from the mobile donors of Rvx. gelatinosus bound either to the RC (pufC) or to the tetraheme subunit (wild type) were similar. The electrons transferred through the reaction center dimer were supplied entirely by the donor pool; their number amounted to about 5 in wild type Rvx. gelatinosus and decreased to 1 in pufC oxidized by ferricyanide. Fluorescence yield was measured as a function of the oxidized fraction of the dimer and its complex shape reveals the contribution of two competing processes: the migration of the excitation energy among the photosynthetic units and the availability of electron donors to the oxidized dimer. The experimental results were simulated and rationalized by a simple kinetic model of the two-electron cycling of the acceptor side combined with aperiodic one-electron redox function of the donor side.

Spectrophotometric Determination of Cysteine Hydrochloride Content by Ferric Chloride-Potassium Ferricyanide.

In the acidic medium, hydrosulfuryl(-SH) in cysteine hydrochloride can reduce Fe3+ to Fe2+, then Fe2+ react with potassium ferricyanide to form KFe[Fe(CN)6](soluble Prussian blue). Prussian blue has a maximum absorption at 727 nm, Bill's law is observed between mass concentration of cysteine hydrochloride and absorbance of Prussian blue, the content of cysteine hydrochloride is indirectly determinated by measuring the absorbance of Prussian blue. An accurate, simple, fast spectrophotometric method for the determination of cysteine hydrochloride content by ferric chloride-potassium ferricyanide has been established. The optimal determination conditions of cysteine hydrochloride content are explored. The cysteine hydrochloride content is determinate by this method.

A possible relationship between the effect of factors on photoactivation of photosystem II depleted of functional Mn and cytochrome b559.

The photoassembly of the Mn4CaO5 cluster in Mn-depleted photosystem II preparations (photoactivation) was studied under the influence of oxidants, reductants and pH. New data on the effect of these factors on the photoactivation yield are presented. The presence of the oxidant, ferricyanide, negatively affected the photoactivation yield over the entire concentration range studied (0-1 mM). In contrast to ferricyanide, the addition of the reductant, ferrocyanide, up to 1 mM resulted in an increase in the photoactivation yield. Other reductants either did not significantly affect (diphenylcarbazide) or suppressed (ascorbate) the photoactivation yield. The effect of ferrocyanide on photoactivation were found to be similar dichlorophenolindophenol. Investigation of the photoactivation yield as a function of pH revealed that the maximum yield was observed at pH 6.5 in the presence of ferrocyanide and DCPIP, and at pH 5.5 without additives. In addition, the photoactivation yield at pH 5.5 was the same without and with the addition of ferrocyanide or dichlorophenolindophenol. Although ferricyanide suppressed the photoactivation, the photoactivation yield increased in the presence of ferricyanide by shifting the pH to the acidic region. The samples contained approximately 25 % of the HP cyt b559, which was in the reduced state, as the absorbance at 559 nm was decreased upon addition of ferricyanide and subsequent addition of ferrocyanide returned the spectrum to the baseline. A possible relationship between the effect of factors on the photoactivation and the involvement of cyt b559 in the protection of PSII from oxidative damage on the donor side is discussed.

Rewiring the respiratory pathway of Lactococcus lactis to enhance extracellular electron transfer.

Lactococcus lactis, a lactic acid bacterium with a typical fermentative metabolism, can also use oxygen as an extracellular electron acceptor. Here we demonstrate, for the first time, that L. lactis blocked in NAD+ regeneration can use the alternative electron acceptor ferricyanide to support growth. By electrochemical analysis and characterization of strains carrying mutations in the respiratory chain, we pinpoint the essential role of the NADH dehydrogenase and 2-amino-3-carboxy-1,4-naphtoquinone in extracellular electron transfer (EET) and uncover the underlying pathway systematically. Ferricyanide respiration has unexpected effects on L. lactis, e.g., we find that morphology is altered from the normal coccoid to a more rod shaped appearance, and that acid resistance is increased. Using adaptive laboratory evolution (ALE), we successfully enhance the capacity for EET. Whole-genome sequencing reveals the underlying reason for the observed enhanced EET capacity to be a late-stage blocking of menaquinone biosynthesis. The perspectives of the study are numerous, especially within food fermentation and microbiome engineering, where EET can help relieve oxidative stress, promote growth of oxygen sensitive microorganisms and play critical roles in shaping microbial communities.

Highly sensitive detection of fluoride based on poly(3-aminophenylboronic acid)-reduced graphene oxide multilayer modified electrode.

A novel electrochemical method for detecting fluoride was developed based on gold electrode modified by layer-by-layer (LBL) assembly of poly(3-aminophenylboronic acid)-reduced graphene oxide (PAPBA-RGO) multilayers. The PAPBA-RGO multilayer-modified gold electrode was constructed by using alternating LBL assembly of RGO and PAPBA on a bare gold electrode by one-step electrodeposition. Fluoride was electrochemically determined based on the proposed modified electrode by evaluating the changes in peak current for potassium ferricyanide reduction caused by the conjunction of fluoride and boronic acid groups of PAPBA. The results indicated that the peak current for potassium ferricyanide reduction obviously decreased with the increasing fluoride concentration. The response range of our method for fluoride was 1 × 10-8 to 1 × 10-1 M with a detection limit of 6 × 10-10 M and high sensitivity and selectivity. This method was applied to detect fluoride in tap water, rice, apple, and edible fungi samples.

Investigation of H2O2 Electrochemical Behavior on Ferricyanide-Confined Electrode Based on Ionic Liquid-Functionalized Silica-Mesostructured Cellular Foam.

In this work, ionic liquid (IL) of 1-propyl-3-methyl imidazolium chloride-functionalized silica-mesostructured cellular foam (MCF) was prepared. The obtained MCF-IL was used to construct the Fe(CN)63--confined electrode (MCF-IL-Fe(CN)63-/PVA) and H2O2 electrochemical behavior on the electrode was investigated. It was found that H2O2 was oxidized on the freshly prepared electrode while catalytically electro-reduced on the acid pretreated one. Cyclic voltametric results revealed that the real catalyst for catalytic reduction of H2O2 was Prussian blue (PB) rather than Fe(CN)63-. The electrocatalytic ability of the acid-pretreated MCF-IL-Fe(CN)63-/PVA electrode offered a wide linear range for H2O2 detection. The present study on H2O2 electrochemical behavior on an MCF-IL-Fe(CN)63-/PVA electrode might provide useful information for further developing integrated Fe(CN)63--mediated biosensors as H2O2 is extensively involved in the classic reaction containing oxidase enzymes.

A Novel Electrochemical Aptasensor Based on a New Platform of Samarium Molybdate Flower-like Nanoparticles@Poly(pyrrole) for Non-Invasive Determination of Saliva CORTISOL.

Cortisol, a famous stress biomarker, can be considered a potential predictor of cardiac diseases in humans. The presence of cortisol in saliva has encouraged researchers to design point-of-care devices for cortisol concentration in biological fluids. Here, human salivary cortisol was analyzed through a new non-invasive voltammetric aptasensor. Although cortisol is an electroactive compound, generally, the reduction in the current peak has been considered; however, this does not show a strong signal on a bare electrode surface, especially at low concentration levels. Hence, in this study, cortisol concentration was measured electrochemically and indirectly by monitoring the difference between electrochemical probe signals in the presence and absence of cortisol. A new polymeric nanocomposite of samarium molybdate flower-like nanoparticles decorated in poly(pyrrole) was electro-synthesized on the surface of a glassy carbon electrode. Then, reduced graphene oxide was cast on the surface. Finally, the cortisol aptamer was immobilized covalently on the reduced graphene oxide. This platform was used to increase the oxidation current peak of the ferricyanide solution as a probe as well as its electrocatalyst. The novel designed polymeric has the potential ability for effective immobilization of aptamers on the electrode surface without decreasing their biological activities. Additionally, it can enhance the probe electrochemical signal. The differential pulse voltammetric method (DPV) was applied as the detection technique. By optimizing the effective parameters, a determination range of 5.0 × 10-14-1.5 × 10-11 mol/L and a limit of detection of 4.5 × 10-14 mol/L were obtained. Selectivity of the proposed aptasensor relative to ß-estradiol, progesterone and also prednisolone was studied as well. Finally, cortisol in a healthy human saliva sample was successfully analyzed by the proposed biosensors.

Photosynthetic microbial fuel cells for methanol treatment using graphene electrodes.

Photosynthetic microbial fuel cells (pMFC) represent a promising approach for treating methanol (CH3OH) wastewater. However, their use is constrained by a lack of knowledge on the extracellular electron transfer capabilities of photosynthetic methylotrophs, especially when coupled with metal electrodes. This study assessed the CH3OH oxidation capabilities of Rhodobacter sphaeroides 2.4.1 in two-compartment pMFCs. A 3D nickel (Ni) foam modified with plasma-grown graphene (Gr) was used as an anode, nitrate mineral salts media (NMS) supplemented with 0.1% CH3OH as anolyte, carbon brush as cathode, and 50 mM ferricyanide as catholyte. Two simultaneous pMFCs that used bare Ni foam and carbon felt served as controls. The Ni/Gr electrode registered a two-fold lower charge transfer resistance (0.005 kΩ cm2) and correspondingly 16-fold higher power density (141 mW/m2) compared to controls. The underlying reasons for the enhanced performance of R. sphaeroides at the graphene interface were discerned. The real-time polymerase chain reaction (PCR) analysis revealed the upregulation of cytochrome c oxidase, aa3 type, subunit I gene, and Flp pilus assembly protein genes in the sessile cells compared to their planktonic counterparts. The key EET pathways used for sustaining CH3OH oxidation were discussed.

Modification of polyacrylonitrile-derived carbon nanofibers and bacteriophages on screen-printed electrodes: A portable electrochemical biosensor for rapid detection of Escherichia coli.

A facile method was developed for fabricating a disposable phage-based electrochemical biosensor for the detection of Escherichia coli. Bare screen-printed electrodes (SPEs) were modified using a two-step drop-casting method, in which polyacrylonitrile-derived electrospun carbon nanofibers (CNFs) were deposited, followed by E. coli bacteriophage immobilization. The deposition of CNFs increased the surface area for bacteriophage immobilization while maintaining a conductive link for ferro/ferricyanide redox transitions. Cyclic voltammetry and electrochemical impedance spectroscopy confirmed that the CNF modification increased the electron-transfer rate, whereas bacteriophages and E. coli blocked electron transfer at the electrode. The biosensor achieved a response within 10 min and a linear response in the E. coli concentration range of 102-106 CFU/mL. A limit of detection (LOD) of 36 CFU/mL in phosphate-buffered saline was achieved, which is the lowest LOD reported thus far for phage-based disposable SPE sensors. The biosensor exhibited recovery rates between 106 % and 119 % for E. coli detection in apple juice. The proposed fabrication method allowed electrodes to be obtained from different production batches with remarkable consistency and reproducibility, and they remained stable at room temperature for one month. Thus, a phage-based disposable SPE that can be used for bacterial detection was developed for the first time.

Potassium ferricyanide oxidizes silicon quantum dots under alkaline conditions to produce chemiluminescence for uric acid detection in human urine.

Potassium ferricyanide (K3 (Fe(CN)6 )) could directly oxidize silicon quantum dots (Si QDs) to generate chemiluminescence (CL) under alkaline conditions. It was noteworthy that in the Si QDs-K3 (Fe(CN)6 )-NaOH CL system, the Si QDs worked as a new luminescent material. In addition, the signal intensity of this CL system could be weakened with the addition of uric acid (UA). Based on these, we exploited a new easy and convenient determination method of UA. This method only needed filtration and dilution of UA, without other pretreatment. The constructed system exhibited a linear relationship that ranged from 0.50 to 4.50 mmol·L-1 , with 0.24 mmol·L-1 of detection limit, and this system had successfully demonstrated the detection of UA in human urine. In addition, this work also broaden the application of the Si QDs in CL research.

Determination of polyanion utilizing a promoted glucose oxidase enzymatic reaction by ε-poly-L-lysine.

In this paper, a sensitive determination method for polyanion using a glucose oxidase (GOx) enzymatic reaction with ferricyanide ion is described. We previously reported that the GOx enzymatic reaction was significantly promoted by a cationic polymer of ε-poly-L-lysine (εPL), and the enzymatic reaction could be utilized for the determination of εPL. Generally, polycation stoichiometrically forms polyion complex with polyanion. Thus, it is expected that the promotion effect of εPL on the enzymatic reaction is interfered by polyanion, and the enzymatic reaction is also applicable to the determination of polyanion. Predictably, the promotion effect of εPL was stoichiometrically interfered by polyanions, such as polyvinyl sulfate and polyacrylate, and the interference effect allowed for the determination of the polyanions. The detection limit of polyanion was estimated to be ~ 0.3 µeq L-1. As a preliminary application, the proposed method was applied to the determination of anionic polymer of heparin in a human plasma.

Elimination of oxygen interference in the photoelectrochemical sensor with ferricyanide shield oxygen reduction for point of care testing.

As a sensitive and promising detection method, photoelectrochemical (PEC) sensor has been widely used in biochemical analysis field. However, the interferences from environment, especially dissolved oxygen, often impact the stability and precision of PEC sensors, which limit its practical applications. Here, we report a dissolved oxygen insensitive PEC sensor based on a proposed indirect electron transfer model. Through the detailed study of the charge transfer process, we determined that the photocurrent mainly comes from the electrochemical reaction between the photochemical products and the electrode, rather than direct charge transfer between the photoelectric materials and the electrode. The newly designed PEC sensor used ferricyanide to shield oxygen reduction and eliminated the influence from variable oxygen solubility. This sensor maintained robust responses over an extremely wide range (1.0-7.5 mg/L) of dissolved oxygen concentrations. To further demonstrate its capability, a smartphone based portable immunosensor was constructed for the detection of human serum albumin (HSA), which exhibited excellent stability and accuracy. The relative error of current was reduced by 81.3% over traditional electron donor solution. This work effectively improves the stability of PEC sensors, and lays the foundation for the subsequently practical applications of PEC sensor in point-of-care testing.

Target amplification-free detection of urinary microRNA for diabetic nephropathy diagnosis with electrocatalytic reaction.

Diabetic nephropathy (DN) is a serious diabetic complication, usually developed from type II diabetes mellitus (T2DM) and known as type II DN (T2DN). New emerging biomarkers for T2DN are microRNAs (miRNAs) which have been studied for the noninvasive early-stage detection of the disease. In this work, a nucleic acid amplification-free miRNA-124 sensor based on target-induced strand displacement on magnetic beads, and by using methylene blue-loaded silica particles as a label was developed. Measurement methods can be either visual observation, spectrophotometry, or electrochemistry. After incubation and separation of the magnetic particles, a blue-violet solution (564 nm) appeared, depending on the concentration of miRNA displaced. For electrochemical detection, methylene blue on the silica served as a redox mediator for the coupled reaction with ferricyanide in the solution phase. At the electrode surface, ferricyanide was re-reduced to ferrocyanide, and was thus available for further reaction with methylene blue, forming an amplification cycle. After optimization, the total assay time was 60 min, and limits of detection were 1 pM, 6 fM, and 0.65 fM, by the naked eye, spectrophotometry and electrochemistry, respectively. The miRNAs in 42 suspected urine samples from patients suffering from either diabetic nephropathy, diabetes mellitus, or chronic kidney disease were validated by comparing with the droplet digital polymerase chain reaction (ddPCR).

A surface protein-imprinted biosensor based on boronate affinity for the detection of anti-human immunoglobulin G.

A surface protein-imprinted biosensor was constructed on a screen-printed carbon electrode (SPCE) for the detection of anti-human immunoglobulin G (anti-IgG). The SPCE was successively decorated with aminated graphene (NH2-G) and gold nanobipyramids (AuNBs) for signal amplification. Then 4-mercaptophenylboric acid (4-MPBA) was covalently anchored to the surface of AuNBs for capturing anti-IgG template through boronate affinity binding. The decorated SPCE was then deposited with an imprinting layer generated by the electropolymerization of pyrrole. After removal of the anti-IgG template by the dissociation of the boronate ester in an acidic solution, three-dimensional (3D) cavities complementary to the anti-IgG template were formed in the imprinting layer of polypyrrole (PPy). The molecularly imprinted polymers (MIP)-based biosensor was used for the detection of anti-IgG, exhibiting a wide linear range from 0.05 to 100 ng mL-1 and a low limit of detection of 0.017 ng mL-1 (S/N = 3). In addition, the MIP-based anti-IgG biosensor also shows high selectivity, reproducibility and stability. Finally, the practicability of the fabricated anti-IgG biosensor was demonstrated by accurate determination of anti-IgG in serum sample.

Low consumption and portable technology for dithionite detection based on potassium ferricyanide differential spectrophotometry method in related advanced oxidation processes.

Carbon neutrality has been received more attention and emerged in wastewater treatment processes. Due to the development of treating technologies with the rising of new-emerging pollutants, the coupled chemical processes also should remain current for the goal of carbon-neutral operation. Among of those updated strategies, several advanced oxidation processes (AOPs) based on dithionite (DTN, S2O42-), a common water treatment agent, have been established for refractory organic contaminations removal. However, in terms of DTN detection, the traditional formol-titration method has several application limits including the low detection sensitivity and high consumption of formaldehyde. In this study, compared with traditional method, a low energy consumption technology has been developed based on the potassium ferricyanide with the carbon consumption decreasing by about 5 times. Moreover, detection limit of DTN (mmol/L level) also was lower than the titration method. The method was established based on the fact that every 1 mol of DTN can react with 2 mol [Fe(CN)6]3- under alkaline condition. According to that potassium ferricyanide (K3 [Fe(CN)6]) has the maximum absorption at 419 nm wavelength, a fitting equation based on the linear relationship between the absorbance variation of K3 [Fe(CN)6] and DTN amount in the ranges of 0-30 µmol with the detection limit of 0.6 µmol was established with the determination coefficient of 0.99935. It was found that there was no obvious influence of the ubiquitous foreign species with the amount lower than 6 mM, 4 mM, 6 mM, 4 mM and 1 mg/L for Cl-, HCO3-, NO3-, SO42- and NOM, respectively. Moreover, methanol and tert-butanol were employed to verify the influence of the presence of organic matters on the determination of DTN and no impact was observed in this study. The proposed method provides a new way for DTN detection with stable and countable performance in the related AOPs with the low electric energy and carbon source consumption and high detection efficiency.