Peptide ligation by chemoselective aminonitrile coupling in water.

Amide bond formation is one of the most important reactions in both chemistry and biology1-4, but there is currently no chemical method of achieving α-peptide ligation in water that tolerates all of the 20 proteinogenic amino acids at the peptide ligation site. The universal genetic code establishes that the biological role of peptides predates life's last universal common ancestor and that peptides played an essential part in the origins of life5-9. The essential role of sulfur in the citric acid cycle, non-ribosomal peptide synthesis and polyketide biosynthesis point towards thioester-dependent peptide ligations preceding RNA-dependent protein synthesis during the evolution of life5,9-13. However, a robust mechanism for aminoacyl thioester formation has not been demonstrated13. Here we report a chemoselective, high-yielding α-aminonitrile ligation that exploits only prebiotically plausible molecules-hydrogen sulfide, thioacetate12,14 and ferricyanide12,14-17 or cyanoacetylene8,14-to yield α-peptides in water. The ligation is extremely selective for α-aminonitrile coupling and tolerates all of the 20 proteinogenic amino acid residues. Two essential features enable peptide ligation in water: the reactivity and pKaH of α-aminonitriles makes them compatible with ligation at neutral pH and N-acylation stabilizes the peptide product and activates the peptide precursor to (biomimetic) N-to-C peptide ligation. Our model unites prebiotic aminonitrile synthesis and biological α-peptides, suggesting that short N-acyl peptide nitriles were plausible substrates during early evolution.

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.

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).

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.

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.

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.

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.

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.

Investigation of the pH-dependence of the oxidation of FAD in VcCRY-1, a member of the cryptochrome-DASH family.

Vibrio cholerae cryptochrome-1 (VcCRY-1) is a member of the cryptochrome DASH family. The flavoprotein appears to use blue light both for repair of cyclobutane pyrimidine dimers (CPDs) on DNA and signal transduction. Earlier, we found that it was almost impossible to oxidize the FADH· state upon binding to a CPD, and, in the absence of substrate, the rate of FADH· oxidation was much larger at high pH (Gindt et al. in Biochemistry 54:2802-2805, 2015). Here, we present the pH-dependence of the oxidation of FADH· by ferricyanide, which revealed a switch between slow and fast oxidation with a pKa ≈ 7.0. Stopped-flow mixing was used to measure the oxidation of FADH- to FADH· at pH 6.7 and 7.5. Substrate binding was required to slow down this oxidation such that it could be measured with stopped flow, but there was only a small effect of pH. In addition, resonance Raman measurements of FADH· in VcCRY-1 at pH 6.5 and 7.5 were performed to probe for structural changes near the FAD cofactor related to the observed changes in rate of FADH· oxidation. Only substrate binding seemed to induce a change near the FAD cofactor that may relate to the change in oxidation kinetics. The pH-effect on the FADH· oxidation rate, which is rate-limited by the proton acceptor, does not seem to be due to a protein structural change near the FAD cofactor. Instead, a conserved glutamate in CRY-DASH may control the deprotonation of FADH· and give rise to the pH-effect.

Miniaturized probe on polymer SU-8 with array of individually addressable microelectrodes for electrochemical analysis in neural and other biological tissues.

An SU-8 probe with an array of nine, individually addressable gold microband electrodes (100 µm long, 4 µm wide, separated by 4-µm gaps) was photolithographically fabricated and characterized for detection of low concentrations of chemicals in confined spaces and in vivo studies of biological tissues. The probe's shank (6 mm long, 100 µm wide, 100 µm thick) is flexible, but exhibits sufficient sharpness and rigidity to be inserted into soft tissue. Laser micromachining was used to define probe geometry by spatially revealing the underlying sacrificial aluminum layer, which was then etched to free the probes from a silicon wafer. Perfusion with fluorescent nanobeads showed that, like a carbon fiber electrode, the probe produced no noticeable damage when inserted into rat brain, in contrast to damage from an inserted microdialysis probe. The individual addressability of the electrodes allows single and multiple electrode activation. Redox cycling is possible, where adjacent electrodes serve as generators (that oxidize or reduce molecules) and collectors (that do the opposite) to amplify signals of small concentrations without background subtraction. Information about electrochemical mechanisms and kinetics may also be obtained. Detection limits for potassium ferricyanide in potassium chloride electrolyte of 2.19, 1.25, and 2.08 µM and for dopamine in artificial cerebral spinal fluid of 1.94, 1.08, and 5.66 µM for generators alone and for generators and collectors during redox cycling, respectively, were obtained.

Calibration curve-free electrochemical quantitation by micro-nano multi-scale gap devices.

Quantitation without relying on the calibration curve has long been an issue of overcoming analytical problems accompanied with the inherent limitations of the calibration curve fitting errors. Here, we report on a calibration curve-free method for electrochemical quantitation based on a multi-scale gap device (MGD). The MGD is an integrated device having a series of interdigitated electrodes (IDE) with micro-to-nano gap distances. The device shows a gap-dependent redox current of the analyte when subjected to the electrochemical cycling between the two facing electrodes of its componential IDEs. Based on the fact that the current increases as the gap distance decreases, the analyte concentration could be directly estimated: the rate of increase in the current was directly proportional to the analyte concentration. The calibration curve was not necessary for the quantitation. The accuracy of this MGD approach was better than that of an IDE collection of the same gap distance, which was deteriorated at the larger gap distances particularly. The MGD-based quantitation of dopamine, potassium ferricyanide, and aminophenol was demonstrated in a relatively broad range of concentrations (100 nM-5 mM).

Impedimetric determination of cortisol using screen-printed electrode with aptamer-modified magnetic beads.

A non-invasive aptamer-based electrochemical biosensor using disposable screen-printed graphene electrodes (SPGEs) was developed for simple, rapid, and sensitive determination of cortisol levels. Selective detection of cortisol based on a label-free electrochemical assay was achieved by specific recognition of the cortisol DNA aptamer (CApt). The CApt was modified with streptavidin magnetic beads (MBs) before simple immobilization onto the electrode surface using a neodymium magnet. The electrochemical behavior of the aptamer-based biosensor was assessed by using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) (vs Ag/AgCl). The specific binding between cortisol and CApt resulted in a decrease in charge transfer resistance (Rct) from EIS using [Fe(CN)6]3-/4- with increasing cortisol concentration. Under optimal conditions, a linear range from 0.10 to 100 ng/mL with a low detection limit (3SD/slope) of 2.1 pg/mL was obtained. Furthermore, the proposed biosensing system exhibited a satisfactory recovery in the range 97.4-109.2% with 5.7-6.6% RSD in spiked artificial human sweat. Regarding the applications of this tool, the aptamer-based biosensor has potential to be a versatile and point-of-care (POC) device for simple, sensitive, selective, disposable, and low-cost cortisol detection.

Electrochemical Characteristics of a DNA Modified Electrode as a Function of Percent Binding.

Electrochemical characteristics of immobilized double-stranded DNA (dsDNA) on a Au electrode were studied as a function of coverage using a home-built optoelectrochemical method. The method allows probing of local redox processes on a 6 µm spot by measuring both differential reflectivity (SEED-R) and interferometry (SEED-I). The former is sensitive to redox ions that tend to adsorb to the electrode, while SEED-I is sensitive to nonadsorbing ions. The redox reaction maxima, Rmax and Δmax from SEED-R and SEED-I, respectively, are linearly proportional to amperometric peak current, Imax. The DNA binding is measured by a redox active dye, methylene blue, that intercalates in dsDNA, leading to an Rmax. Concomitantly, the absence of Δmax for [Fe(CN)6]4-/3- by SEED-I ensures that there is no leakage current from voids/defects in the alkanethiol passivation layer at the same spot of measurement. The binding was regulated electrochemically to obtain the binding fraction, f, ranging about three orders of magnitude. A remarkably sharp transition, f = fT = 1.25 × 10-3, was observed. Below fT, dsDNA molecules behaved as individual single-molecule nanoelectrodes. Above the crossover transition, Rmax, per dsDNA molecule dropped rapidly as f-1/2 toward a planar-like monolayer. The SEED-R peak at f ∼ 3.3 × 10-4 (∼270 dsDNA molecules) was (statistically) robust, corresponding to a responsivity of ∼0.45 zeptomoles of dsDNA/spot. Differential pulse voltammetry in the single-molecule regime estimated that the current per dsDNA molecule was ∼4.1 fA. Compared with published amperometric results, the reported semilogarithmic dependence on target concentration is in the f > fT regime.

Screen-Printed Carbon Electrodes Modified with Graphene Oxide for the Design of a Reagent-Free NAD+-Dependent Biosensor Array.

A facile approach for the construction of reagent-free electrochemical dehydrogenase-based biosensors is presented. Enzymes and cofactors (NAD+ and Fe(CN)63-) were immobilized by modification of screen-printed carbon electrodes with graphene oxide (GO) and an additional layer of cellulose acetate. The sensor system was exemplarily optimized for an l-lactate electrode in terms of GO concentration, working potential, and pH value. The biosensor exhibited best characteristics at pH 7.5 in 100 mM potassium phosphate buffer at an applied potential of +0.250 V versus an internal pseudo Ag reference electrode. Thereby, sensor performance was characterized by a linear working range from 0.25 to 4 mM and a sensitivity of 0.14 µA mM-1. The detection principle was additionally evaluated with three other dehydrogenases (d-lactate dehydrogenase, alcohol dehydrogenase, and formate dehydrogenase, respectively). The developed reagentless biosensor array enabled simultaneous and cross-talk free determination of l-lactate, d-lactate, ethanol, and formate.

C60 Mediated Ion Pair Interaction for Label-Free Electrochemical Immunosensing with Nanoporous Anodic Alumina Nanochannels.

Label-free biosensing based on the nanoporous anodic alumina (NAA) membrane emerged as a versatile biosensing platform in the recent decade. In the present work, we developed a new immunosensing strategy based on the nanochannels of NAA and the ion pair interaction mediated by electrochemistry of C60. The NAA served as the matrix for the immobilization of the capture antibodies. The incubation of target antigens resulted in the formation of the immunocomplexes and thus an increase of the steric hindrance of the nanochannels. Therefore, the concentration of the redox probe transported through the nanochannels decreases, which can be detected at the working electrode modified with C60. Herein, we initially found that the cathodic peak ascribed to the reduction of C60 to C60- was obviously enhanced by the presence of the redox probe K3[Fe(CN)6] and which was contributed to the formation of a ternary ion association complex among C60, tetraoctylammonium bromide, and K3[Fe(CN)6]. Therefore, the transportation of K3[Fe(CN)6] though the NAA-based bionanochannels can be detected by a C60 modified electrode with an amplified signal. Choosing human epididymis protein 4 (HE4) as the model target, a linear range of 1.0 ng mL-1 to 100 ng mL-1 can be established between the peak current obtained from the differential pulse voltammetric response of the platform and the concentration of HE4. The detection limit was 0.2 ng mL-1. This study not only provides a new avenue to develop the other nanochannel-based biosensing platform for a variety of other disease biomarkers but also contributes to the electrochemistry of fullerene.

Label-Free and Immobilization-Free Electrochemical Magnetobiosensor for Sensitive Detection of 5-Hydroxymethylcytosine in Genomic DNA.

DNA 5-hydroxymethylcytosine (5-hmC) is an important epigenetic biomarker for tumorigenesis, and the loss of 5-hmC levels is associated with leukemia and melanoma cancers. However, it is a great challenge to discriminate 5-hmC from 5-methylcytosine (5-mC) using the conventional bisulfite conversion methods. Herein, we report a label-free and immobilization-free electrochemical magnetobiosensor for sensitive quantification of 5-hmC in genomic DNA based on a dual signal amplification strategy coupled with terminal deoxynucleotidyl transferase (TDT) enzymatic amplification and Ru(III) redox cycling. This screen-printed carbon electrode (SPCE)-based electrochemical magnetobiosensor shows distinct advantages of having low cost and simple fabrication and being label-free, immobilization-free, PCR-free, and radioactive-free. It exhibits high sensitivity with a detection limit of as low as 9.06 fM and a large dynamic range from 0.01 to 1000 pM. Importantly, this biosensor can discriminate 5-hmC from cytosine and 5-mC, and it can successfully detect 5-hmC in live cells.

Immobilized bacterial biosensor for rapid and effective monitoring of acute toxicity in water.

The use of biosensors by using microorganisms such as bacteria have short life cycles and provide other advantages. One colorimetric biosensor technique that has been developed is the use of a biosensor utilizing the incorporation of Prussian blue formation reactions mediated by E. coli bioreactors with ferricyanide. Immobilization is a method that allows the bacteria can be used for long-term without reducing its ability as bioreceptor. This study aimed to develop a novel and rapid immobilized bacterial biosensor for the detection of toxic compound in water and to evaluate their analytical performances. Immobilization of E. coli performed by trapping method using alginate material support. The bacterial suspension was mixed with sodium alginate (1:1 v/v), and the mixture was continuously dropped in CaCl2 solution to be a form of beads. The beads were used as bioreceptor to detect toxicants regarding cadmium, arsenic, mercury, chromium and lead solutions with Prussian blue as a colorimetric indicator. The linearity and sensitivity of detection of beads to the toxicants were tested, the stability of repeated use and storage were evaluated as well. The results showed that E. coli could be immobilized using alginate with response value was correlated with toxic concentration. The developed biosensor was more stable when used repeatedly and could be stored in a long time. The immobilization of E. coli in calcium alginate bead was successfully performed as a biosensor system for monitoring acute toxicity in water.

Electrochemical selection of a DNA aptamer, and an impedimetric method for determination of the dedicator of cytokinesis 8 by self-assembly of a thiolated aptamer on a gold electrode.

The autosomal recessive-hyper immunoglobulin E syndromes (AR-HIES) are inherited inborn primary immunodeficiency disorders caused mainly by mutations in the dedicator of cytokinesis 8 (DOCK8) gene. A method is described for the selection of DNA aptamers against DOCK8 protein. The selection was performed by using a gold electrode as the solid matrix for immobilization of DOCK8. This enables voltammetric monitoring of the bound DNA after each selection cycle. After eight rounds of selection, high affinity DNA aptamers for DOCK8 were identified with dissociation constants (Kds) ranging from 3.3 to 66 nM. The aptamer which a Kd of 8.8 nM was used in an aptasensor. A gold electrode was modified by self-assembly of the thiolated aptamer, and the response to the DOCK8 protein was detected by monitoring the change in the electron transfer resistance using the ferro/ferricyanide system as a redox probe. The aptasensor works in the 100 pg.mL-1 to 100 ng.mL-1 DOCK8 concentration range, has a detection limit of 81 pg.mL-1 and good selectivity over other proteins in the serum. Graphical abstractSchematic representation of an electrochemical screening protocol for the selection of DNA aptamer against dedicator of cytokinesis 8 protein using electrode as solid support for target immobilization.

Effect of Poly-l-Lysine Polycation on the Glucose Oxidase/Ferricyanide Composite-Based Second-Generation Blood Glucose Sensors.

Second-generation glucose biosensors are presently the mainstream commercial solution for blood glucose measurement of diabetic patients. Screen-printed carbon electrodes (SPCEs) are the most-used substrate for glucose testing strips. This study adopted hydrophilic and positively charged α-poly-l-lysine (αPLL) as the entrapment matrix for the immobilization of negatively charged glucose oxidase (GOx) and ferricyanide (FIC) on SPCEs to construct a disposable second-generation glucose biosensor. The αPLL modification is shown to facilitate the redox kinetics of FIC and ferrocyanide on the SPCEs. The SPCEs coated with 0.5 mM GOx, 99.5 mM FIC, and 5 mM αPLL had better sensitivity for glucose detection due to the appreciable effect of protonated αPLL on the promotion of electron transfer between GOx and FIC. Moreover, the SPCEs coated with 0.5 mM GOx, 99.5 mM FIC, and 5 mM αPLL were packaged as blood glucose testing strips for the measurement of glucose-containing human serum samples. The glucose testing strips had good linearity from 2.8 mM to 27.5 mM and a detection limit of 2.3 mM. Moreover, the 5 mM αPLL-based glucose testing strips had good long-term stability to maintain GOx activity in aging tests at 50 °C.

Hydrogen Tunnelling as a Probe of the Involvement of Water Vibrational Dynamics in Aqueous Chemistry?

Our study of tunnelling in proton-coupled electron transfer (PCET) oxidation of ascorbate with hexacyanoferrate(III) follows the insights obtained from ultrafast 2D IR spectroscopy and theoretical studies of the vibrational water dynamics that led to the proposal of the involvement of collective intermolecular excitonic vibrational water dynamics in aqueous chemistry. To test the proposal, the hydrogen tunnelling modulation observed in the PCET reaction studied in the presence of low concentrations of various partial hydrophobic solutes in the water reaction system has been analyzed in terms of the proposed involvement of the collective intermolecular vibrational water dynamics in activation process in the case. The strongly linear correlation between common tunnelling signatures, isotopic values of Arrhenius prefactor ratios ln AH/AD and isotopic differences in activation enthalpies ΔΔH‡ (H,D) observed in the process in fairly diluted water solutions containing various partial hydrophobic solutes (such as dioxane, acetonitrile, ethanol, and quaternary ammonium ions) points to the common physical origin of the phenomenon in all the cases. It is suggested that the phenomenon can be rooted in an interplay of delocalized collective intermolecular vibrational dynamics of water correlated with vibrations of the coupled transition configuration, where the donor-acceptor oscillations, the motions being to some degree along the reaction coordinate, lead to modulation of hydrogen tunnelling in the reaction.