Glucose Oxidase Research Articles

Honey induces changes in the molecular structure and microstructure of gluten in wheat-rye sourdoughs.

Chemical oxidizers and redox enzymes have traditionally been used to enhance the quality of baked goods. However, consumers now seek natural and clean-label ingredients, avoiding those with chemical-sounding names. Honey, a natural source of glucose oxidase (GOX), represents a promising alternative to purified enzymes for baking purposes. This study aimed to evaluate the effect of honey on the molecular structure and microstructure of gluten proteins in sourdough fermented by different lactic acid bacteria (LAB) strains. Four wheat-rye (1:1) sourdoughs were prepared, each supplemented with honey and inoculated with a different LAB strain. Additionally, two uninoculated doughs, one with honey (honey dough) and the other without (control dough), were prepared under identical conditions. Electronic paramagnetic resonance spectroscopy revealed the presence of hydrogen peroxide in honey solutions, indicating its role as an active source of GOX. Raman spectroscopy showed that honey addition altered the molecular structure of gluten by increasing the proportion of random coils at the expense of α-helix structures. This change is likely attributed to the competition between honey sugars and gluten proteins for water molecules in this system. Moreover, honey led to a decrease in the free sulfhydryl content of gluten compared to the control dough, suggesting an increase in disulfide crosslinking points. These enhanced protein-protein interactions were observed in scanning electron microscopy micrographs as a coarse gluten network composed of interconnected strands and fibrils. All LAB strains exhibited optimal acidification (pH < 4.3) in honey-supplemented sourdoughs, promoting the hydrolysis of gluten proteins into smaller fragments. Overall, honey-supplemented sourdoughs showed a gradual increase in the β-sheet content while decreasing the proportion of random coils over time. This trend suggests that the polypeptide fragments interacted through interchain hydrogen bonds, leading to a more ordered structure, which likely contributes to providing dough with good baking aptitude.

Fabrication of Multiple-Channel Electrochemical Microneedle Electrode Array via Separated Functionalization and Assembly Method

Real-time monitoring of physiological indicators inside the body is pivotal for contemporary diagnostics and treatments. Implantable electrodes can not only track specific biomarkers but also facilitate therapeutic interventions. By modifying biometric components, implantable electrodes enable in situ metabolite detection in living tissues, notably beneficial in invasive glucose monitoring, which effectively alleviates the self-blood-glucose-managing burden for patients. However, the development of implantable electrochemical electrodes, especially multi-channel sensing devices, still faces challenges: (1) The complexity of direct preparation hinders functionalized or multi-parameter sensing on a small scale. (2) The fine structure of individual electrodes results in low spatial resolution for sensor functionalization. (3) There is limited conductivity due to simple device structures and weakly conductive electrode materials (such as silicon or polymers). To address these challenges, we developed multiple-channel electrochemical microneedle electrode arrays (MCEMEAs) via a separated functionalization and assembly process. Two-dimensional microneedle (2dMN)-based and one-dimensional microneedle (1dMN)-based electrodes were prepared by laser patterning, which were then modified as sensing electrodes by electrochemical deposition and glucose oxidase decoration to achieve separated functionalization and reduce mutual interference. The electrodes were then assembled into 2dMN- and 1dMN-based multi-channel electrochemical arrays (MCEAs), respectively, to avoid damaging functionalized coatings. In vitro and in vivo results demonstrated that the as-prepared MCEAs exhibit excellent transdermal capability, detection sensitivity, selectivity, and reproducibility, which was capable of real-time, in situ glucose concentration monitoring.

Screening of Lactic Acid Bacteria Strains for Potential Sourdough and Bread Applications: Enzyme Expression and Exopolysaccharide Production.

Twenty-eight strains of lactic acid bacteria (LAB) were characterized for the ability to express enzymes of interest (including protease, xylanase, α-amylase, laccase, and glucose oxidase) as well as the ability to produce exopolysaccharide (EPS). The screening of enzyme capability for all LAB strains proceeded in a progressive 3-stage manner that helps to profile the efficiency of LAB strains in expressing chosen enzymes (Stage 1), highlights the strains with affinity for flour as the substrate (Stage 2), and discerns strains that can adapt well in a simulated starter environment (Stage 3). The theoretical ability of LAB to express these enzymes was also assessed using Basic Local Alignment Search Tool (BLAST) analysis to identify the underlying genes in the whole genome sequence. By consolidating both experimental data and information obtained from BLAST, three LAB strains were deemed optimal in expressing enzymes, namely, Lb. delbrueckii subsp. bulgaricus (RBL 52), Lb. rhamnosus (RBL 102), and Lb. plantarum (ATCC 10241). Meanwhile, EPS-producing capabilities were observed for 10 out of 28 LAB strains, among which, Lactococcus lactis subsp. diacetylactis (RBL 37) had the highest total EPS yield (274.15mg polysaccharide/L culture) and produced 46.2% polysaccharide with a molecular mass of more than 100kDa.

Evaluation of vat blue removal by Aspergillus niger using scanning electron microscopy and glucose oxidase enzyme activity

AbstractThe use of fungi to reduce dye removal in water from diverse industries can be an efficient option for solving environmental problems. However, the mycoremediation processes involved in color removal are unknown. In this article, Aspergillus niger CDBB‐H‐175 was applied for vat blue discoloration (DC). Hence, inactive biomass by two methods (metabolic inhibitor and heat) and active biomass were studied by scanning electron microscopy and enzyme activities to determine the vat blue DC. When using active biomass, 62% of DC, glucose oxidase (GOX) enzyme activity, 1.19 U/mL and 1.43 mg/L of H2O2 were obtained. It was observed that 68% of DC was due to the degradation process and 32% was due to biomass sorption. When the biomass was inactivated with sodium azide, DC increased by 90% (1.45‐fold), low GOX activity (0.138 U/mL), and H2O2 (0.25 mg/L) were detected. High DC might be due to a high dye sorption in hyphae, since an increase in the thickness and morphology of the hyphae were observed by SEM. Inactivated biomass through heating, showed a 69.5% of DC, where 0.01 U/mL GOX and 0.25 mg/L H2O2 were reached, demonstrating that inactivation of A. niger biomass by sodium azide can be considered as an alternative treatment for colored wastewater.

Proportion of Newly Detected Hypothyroidism among the Diabetic and Nondiabetic Pregnant Ladies and Correlation of Different Parameters with Thyroid Stimulatory Hormone in Pregnancy Attending a Tertiary Care Hospital

ABSTRACT Background: Subclinical hypothyroidism and gestational diabetes are the two most common endocrinopathy encountered in daily clinical practice. Many literature showed a higher frequency of hypothyroidism in the persons with diabetes but others did not find it. Objectives: The study was intended to compare the proportion of newly detected hypothyroidism in persons with diabetes and without diabetes in pregnancy (DIP). Materials and Methods: This cross-sectional, observational study enrolled 175 pregnant women by convenient sampling from the Department of Obstetrics and Gynecology, BIRDEM General Hospital, with or without diabetes after IRB clearance. According to the study protocol, Thyroid function test (TFT) and oral glucose tolerance test were done in the participants. Thyroid stimulatory hormone (TSH), free thyroxin, and antithyroid antibodies were tested using the chemiluminescent technique by the ADVIA centaur, Siemens. Plasma glucose was analyzed by Advia-1800, SIEMENS, USA by glucose oxidase method. Data were analyzed by SPSS version 25. Nonparametric tests were chosen for statistical analysis. Results: The median maternal age was 30 years, and half of the participants had a family history of diabetes mellitus (DM). Hundred patients had diabetes either gestational DM or DIP or preexisting diabetes. Among the persons with diabetes, 34% had hypothyroidism, and in euglycemic patients, 29.3% had hypothyroidism. There was no statistically significant (P = 0.785) difference in the median values of TSH across the glycemic status among the hypothyroid subjects. TSH had a positive correlation with thyroid peroxidase antibody (P = 0.174) within the hypothyroid patients. In the study subjects, TSH was inversely correlated with hemoglobin (P &lt; 0.001). Conclusions: There is no statistically significant concordance in diagnosing hypothyroidism in pregnancy between the persons with diabetes and without diabetes.

A Multifunctional Nanoreactor‐Induced Dual Inhibition of HSP70 Strategy for Enhancing Mild Photothermal/Chemodynamic Synergistic Tumor Therapy

AbstractMild photothermal therapy (PTT) is a spatiotemporally controllable method that utilizes the photothermal effect at relatively low temperatures (40‐45 °C) to especially eliminate tumor tissues with negligible side effects on the surrounding normal tissues. However, the overexpression of heat shock protein 70 (HSP70) and limited effect of single treatment drastically impeded the therapeutic efficacy. Herein, the constructed multifunctional core‐shell structured Ag‐Cu@SiO2‐PDA/GOx nanoreactors (APG NRs) that provide a dual inhibition of HSP70 strategy for the second near‐infrared photoacoustic (NIR‐II PA) imaging‐guided combined mild PTT/chemodynamic therapy (CDT). The Ag‐Cu cores can convert endogenous H2O2 to hydroxyl radical (•OH), which could induce lipid peroxidation (LPO) and further degrade HSP70. The polydopamine (PDA)/glucose oxidase (GOx) shells were utilized as the NIR‐II photothermal agent to generate low temperature, and the GOx could reduce the energy supplies and inhibit energy‐dependent HSP70 expression. Furthermore, both the generation of •OH and GOx‐mediated energy shortage can reduce HSP70 expression to sensitize mild PTT under 1064 nm laser, and in turn, GOx and laser self‐amplify the catalytic reactions of APG NRs for more production of •OH. The multifunctional nanoreactors will provide more potential possibilities for the clinical employment of mild PTT and the advancement of tumor combination therapies.This article is protected by copyright. All rights reserved

Radiotherapy-sensitized cancer immunotherapy via cGAS-STING immune pathway by activatable nanocascade reaction

Radiotherapy-induced immune activation holds great promise for optimizing cancer treatment efficacy. Here, we describe a clinically used radiosensitizer hafnium oxide (HfO2) that was core coated with a MnO2 shell followed by a glucose oxidase (GOx) doping nanoplatform (HfO2@MnO2@GOx, HMG) to trigger ferroptosis adjuvant effects by glutathione depletion and reactive oxygen species production. This ferroptosis cascade potentiation further sensitized radiotherapy by enhancing DNA damage in 4T1 breast cancer tumor cells. The combination of HMG nanoparticles and radiotherapy effectively activated the damaged DNA and Mn2+-mediated cGAS-STING immune pathway in vitro and in vivo. This process had significant inhibitory effects on cancer progression and initiating an anticancer systemic immune response to prevent distant tumor recurrence and achieve long-lasting tumor suppression of both primary and distant tumors. Furthermore, the as-prepared HMG nanoparticles “turned on” spectral computed tomography (CT)/magnetic resonance dual-modality imaging signals, and demonstrated favorable contrast enhancement capabilities activated by under the GSH tumor microenvironment. This result highlighted the potential of nanoparticles as a theranostic nanoplatform for achieving molecular imaging guided tumor radiotherapy sensitization induced by synergistic immunotherapy.

Inhibiting the SARM1-NAD+ axis reduces oxidative stress-induced damage to retinal and nerve cells

Retinal neurodegenerative diseases are a category of refractory blinding eye conditions closely associated with oxidative stress induced by mitochondrial dysfunction in retinal cells. SARM1, a core driver molecule leading to axonal degeneration, possesses NAD+ enzyme (NADase) activity. However, the role of the SARM1-NAD+ axis in oxidative stress-induced retinal cell death remains unclear. Here, we employed the SARM1 NADase inhibitor DSRM-3716 and established a glucose oxidase (GOx)-induced oxidative stress cell model. We found that compared to the GOx group, the DSRM-3716 pre-treated group reduced the hydrolysis of NAD+, inhibited the elevation of oxidative stress markers induced by GOx, decreased mitochondrial dysfunction, lowered the phosphorylation level of JNK, and attenuated the occurrence of pyroptosis in retinal and nerve cells, thereby providing protection for neurite growth. Further utilization of the JNK activator Anisomycin activated JNK, revealed that the JNK/c-Jun pathway down-regulated NMNAT2 expression. Consequently, it reduced cellular NAD+ synthesis, exacerbated mitochondrial dysfunction and cell pyroptosis, and reversed the protective effect of DSRM-3716 on cells. In summary, the inhibition of SARM1 NADase activity substantially mitigates oxidative damage to retinal cells and mitochondrial damage. Additionally, JNK simultaneously serves as both an upstream and downstream regulator in the SARM1-NAD+ axis, regulating retinal cell pyroptosis and neurite injury. Thus, this study provides new insights into the pathological processes of retinal cell oxidative stress and identifies potential therapeutic targets for retinal neurodegenerative diseases.

Enhancing Substrate Channeling with Multi-Enzyme Architectures in Hydrogen-Bonded Organic Framework.

Hydrogen-bonded organic frameworks (HOF) represent an emerging category of organic structures with high crystallinity and metal-free, which are not commonly observed in alternative porous organic frameworks. These needle-like porous structure can help in stabilizing enzymes and allow transfer of molecules between enzymes participating in cascade reactions for enhanced substrate channelling. Herein, we systematically synthesized and investigated the stability of HOF at extreme conditions followed by one-pot encapsulation of single and bi-enzyme systems. Firstly, we observed HOF to be stable at pH 1 to 14 and at high temperatures (up to 115°C). Secondly, the encapsulated glucose oxidase enzyme (GOX) showed 80 % and 90% of its original activity at 70 °C and pH 11, respectively. Thirdly, transient time close to 0 seconds was observed for HOF encapsulated bi-enzyme cascade reaction system demonstrating a 4.25-fold improvement in catalytic activity when compared to free enzymes with enhanced substrate channelling. Our findings showcase a facile system synthesized under ambient conditions to encapsulate and stabilize enzymes at extreme conditions.

All-printed wearable biosensor based on MWCNT-iron oxide nanocomposite ink for physiological level detection of glucose in human sweat

Wearable sensors for sweat glucose monitoring are gaining massive interest as a patient-friendly and non-invasive way to manage diabetes. The present work offers an alternative on-body method employing an all-printed flexible electrochemical sensor to quantify the amount of glucose in human sweat. The working electrode of the glucose sensor was printed using a custom-formulated ink containing multi-walled carbon nanotube (MWCNT), poly (3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOPT: PSS), and iron (II, III) oxide (Fe3O4) nanoparticles. This novel ink composition has good conductivity, enhanced catalytic activity, and excellent selectivity. The working electrode was modified using Prussian blue (PB) nanoparticles and glucose oxidase enzyme (GOx). The sensor displayed a linear chronoamperometric response to glucose from 1 μM to 400 μM, with a precise detection limit of ∼0.38 μM and an impressive sensitivity of ∼4.495 μAμM−1cm−2. The sensor stored at 4 °C exhibited excellent stability over 60 days, high selectivity, and greater reproducibility. The glucose detection via the standard addition method in human sweat samples acquired a high recovery rate of 96.0–98.6%. Examining human sweat during physical activity also attested to the biosensor's real-time viability. The results also show an impressive correlation between glucose levels obtained from a commercial blood glucose meter and sweat glucose concentrations. Remarkably, the present results outperform previously published printed glucose sensors in terms of detection range, low cost, ease of manufacturing, stability, selectivity, and wearability.

A Biomineralized Bifunctional Patient-Friendly Nanosystem for Sustained Glucose Monitoring and Control in Diabetes.

Regular blood glucose monitoring and control is necessary for people with type 1 or advanced type 2 diabetes, yet diagnosing and treating patients with diabetes in an accurate, sustained and patient-friendly manner remains limited. Here, a glucose-responsive bifunctional nanosystem (PGOxMns) is constructed via one-pot biomineralisation of manganese dioxide with glucose oxidase and ε-poly-L-lysine. Under hyperglycaemic conditions, the cascade reactions that occur when glucose interacts with PGOxMns can trigger the production of Mn(II), which enhances the magnetic resonance imaging signal. Simultaneously, manganese dioxide catalyses the decomposition of toxic hydrogen peroxide into oxygen, which also maintains glucose oxidase (GOx)activity. In an in vivo model of diabetes, PGOxMns is used to monitor glucose levels (0-20mm) and allowed identification of diabetic mice via T1-weighted MRI. Furthermore, PGOxMns is found to have a high insulin-loading capacity (83.6%), likely due to its positive charge. A single subcutaneous injection of insulin-loaded nanosystem (Ins-PGOxMns) into diabetic mice resulted in a rapid and efficient response to a glucose challenge and prolonged blood glucose level control (< 200mgdL-1) for up to 50h. Overall, this proof-of-concept study demonstrates the feasibility of using biomineralised nanosystems to develop patient-friendly strategies for glucose monitoring and control.

A Zn-MOF-GOx-based Cascade Nanoreactor Promotes Diabetic Infected Wound Healing by NO Release and Microenvironment Regulation

Diabetic wound healing is a great clinical challenge due to the microenvironment of hyperglycemia and high pH value, bacterial infection and persistent inflammation. Here, we develop a cascade nanoreactor hydrogel (Arg@Zn-MOF-GOx Gel, AZG-Gel) with arginine (Arg) loaded Zinc metal organic framework (Zn-MOF) and glucose oxidase (GOx) based on chondroitin sulfate (CS) and Pluronic (F127) to accelerate diabetic infected wound healing. GOx in AZG-Gel was triggered by hyperglycemic environment to reduce local glucose and pH, and simultaneously produced hydrogen peroxide (H2O2) to enable Arg to release nitric oxide (NO) for inflammation regulation, providing a suitable microenvironment for wound healing. Zinc ions (Zn2+) released from acid-responsive Zn-MOF significantly inhibited the proliferation and biofilm formation of S.aureus and E.coli. AZG-Gel significantly accelerated diabetic infected wound healing by down-regulating pro-inflammatory tumor necrosis factor (TNF)-α and interleukin (IL)-6, up-regulating anti-inflammatory factor IL-4, promoting angiogenesis and collagen deposition in vivo. Collectively, our nanoreactor cascade strategy combining “endogenous improvement (reducing glucose and pH)” with “exogenous resistance (anti-bacterial and anti-inflammatory)” provides a new idea for promoting diabetic infected wound healing by addressing both symptoms and root causes. Statement of SignificanceA cascade nanoreactor (AZG-Gel) is constructed to solve three key problems in diabetic wound healing, namely, hyperglycemia and high pH microenvironment, bacterial infection and persistent inflammation. Local glucose and pH levels are reduced by GOx to provide a suitable microenvironment for wound healing. The release of Zn2+ significantly inhibits bacterial proliferation and biofilm formation, and NO reduces wound inflammation and promotes angiogenesis. The pH change when AZG-Gel is applied to wounds is expected to enable the visualization of wound healing to guide the treatment of diabetic wound. Our strategy of “endogenous improvement (reducing glucose and pH)” combined with “exogenous resistance (anti-bacterial and anti-inflammatory)” provides a new way for promoting diabetic wound healing.

Chitosan-based GOx@Co-MOF composite hydrogel: A promising strategy for enhanced antibacterial and wound healing effects

A novel composite hydrogel was synthesized via Schiff base reaction between chitosan and di-functional poly(ethylene glycol) (DF-PEG), incorporating glucose oxidase (GOx) and cobalt metal-organic frameworks (Co-MOF). The resulting CS/PEG/GOx@Co-MOF composite hydrogel was characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA), and energy-dispersive X-ray spectroscopy (EDS). The results confirmed successful integration and uniform distribution of Co-MOF within the hydrogel matrix. Functionally, the hydrogel exploits the catalytic decomposition of glucose by GOx to generate gluconic acid and hydrogen peroxide (H2O2), while Co-MOF gradually releases metal ions and protects GOx. This synergy enhanced the antibacterial activity of the composite hydrogel against both Gram-positive (S. aureus) and Gram-negative bacteria (E. coli), outperforming conventional chitosan-based hydrogels. The potential of the composite hydrogel in treating wound infections was evaluated through antibacterial and wound healing experiments. Overall, CS/PEG/GOx@Co-MOF hydrogel holds great promise for the treatment of wound infections, paving the way for further research and potential clinical applications.

Construction of multi(iron)porphyrin array multilayers on quartz surface as peroxidase mimetics for H2O2 and glucose colorimetric, fluorometric and color-read detection

Multi(metallo)porphyrin arrays are important artificial biomimetics for mimicking the biological catalytic reactions in nature. Here, we report construction of multi(iron)porphyrin array ultrathin films on the quartz surface by coordination-driven layer-by-layer (LBL) assembling method with the Pd(II) ion as a connector and iron(III) tetrapyridylporphyrin (FeTPyP) as a linker. It is revealed that the (Pd-FeTPyP)n LBL multilayers can catalyze oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H2O2 with peroxidase-like activity, resulting in the formation of the blue-colored oxidized TMB (oxTMB). Coupled with glucose oxidase, three sensitive probes based on the colorimetric, fluorometric and color-read methods have been developed by using modified quartz plates as the heterogenous catalyst, with a glucose detection limit of about 0.89, 1.5 and 0.65 μM, respectively. Mechanism studies confirm an enhanced affinity between (Pd-FeTPyP)n and H2O2/TMB molecules and an improved catalytic performance, which are attributed to the reason that the FeTPyP macrocycles are well-dispersed and arranged on the two-dimensional quartz surface. Furthermore, owing to chemically anchored on the transparent quartz surface, the (Pd-FeTPyP)n films exhibit good stability with naked-eye observation of color change under moderate conditions, thus providing a potential candidate for developing colorimetric, fluorometric and color-read glucose biosensors.

An intra-stimuli resonance energy transfer-based electrochemiluminescence biosensor for DNA methylation detection

Aberrant DNA methylation, as one of the promising biomarkers, is intimately connected to a variety of genetic and tumor diseases. Herein, we constructed an innovative intra-stimuli electrochemiluminescence resonance energy transfer (ECL-RET) biosensor for methylated DNA detection with high sensitivity and accuracy. Glucose oxidase (GOD) was modified at the terminal of assist DNA as functional conjugation (assist DNA-GOD), which then hybridized with methylated target DNA and hairpin DNA to form Y-shaped DNA ternary conjugate. Ru(bpy)32+ encapsulated silica nanospheres (RuSiO2 NPs) were uniformly coated with silver nanoparticles (Ag NPs) and the formed RuSiO2@Ag nanocomposites were introduced to the terminal of hairpin DNA via amide bond. The combination of the ECL donor (RuSiO2 NPs) and acceptor (Ag NPs) on one nanostructure yielded an intra-molecular ECL-RET. Afterward, the presence of H2O2 as the product of GOD catalyzing glucose induced the etching of Ag NPs and subsequent recovery of ECL signal from RuSiO2 NPs. Thus, due to the dependence of the ECL enhancement on the target concentration, this method achieved the quantitative analysis of methylated target DNA with good selectivity, repeatability, and practical utility.

Dual-Catalysts-Embedded spontaneously propelling asymmetric micromotors using triple emulsion microfluidic synthesis for highly efficient nano/microplastic removal

Plastics, which are pivotal in modern technological progress and have a broad range of applications, now pose a significant environmental and health risk globally. Despite their detrimental environmental effects, there is a lack in research on effective plastic removal and treatment strategies. Addressing this urgent issue necessitates the development of innovative and efficient materials and methods for removing plastics from contaminated environments. In this study, we utilized a microfluidic technique to create novel and eco-friendly asymmetrically structured micromotors made of glucose oxidase (GOx)/catalase (Cat) embedded in polyethylene glycol diacrylate (PEGDA). These micromotors are specifically engineered to efficiently remove substantial amounts of plastics from ecosystems. The microfluidic process enabled the production of micromotors with uniform yet asymmetric shapes through aqueous two-phase separation and UV polymerization. By incorporating functional nanomaterials, these micromotors can be customized for specific wastewater treatment needs. Our design utilizes GOx and Cat to consume glucose as fuel, generating oxygen microbubbles. These bubbles not only propel the micromotors but also capture suspended plastic nano/micro-particles, effectively removing them from aquatic environments. The high surface tension of the bubbles allows for the entrapment of plastics at their interface, facilitating their removal from the water. In addition, non-toxic nature of these GOx/Cat PEGDA micromotors makes them especially suitable for environmental remediation and biological applications, addressing the urgent need for environmentally benign solutions to plastic pollution.

Optimized Fabrication of Carbon-Fiber Microbiosensors for Codetection of Glucose and Dopamine in Brain Tissue.

Dopamine (DA) signaling is critically important in striatal function, and this metabolically demanding process is fueled largely by glucose. However, DA and glucose are typically studied independently and, as such, the precise relationship between DA release and glucose availability remains unclear. Fast-scan cyclic voltammetry (FSCV) is commonly coupled with carbon-fiber microelectrodes to study DA transients. These microelectrodes can be modified with glucose oxidase (GOx) to generate microbiosensors capable of simultaneously quantifying real-time and physiologically relevant fluctuations of glucose, a nonelectrochemically active substrate, and DA, which is readily oxidized and reduced at the electrode surface. A chitosan hydrogel can be electrodeposited to entrap the oxidase enzyme on the sensor surface for stable, sensitive, and selective codetection of glucose and DA using FSCV. This strategy can also be used to entrap lactate oxidase on the carbon-fiber surface for codetection of lactate and DA. However, these custom probes are individually fabricated by hand, and performance is variable. This study characterizes the physical nature of the hydrogel and its effects on the acquired electrochemical data in the detection of glucose (2.6 mM) and DA (1 μM). The results demonstrate that the electrodeposition of the hydrogel membrane is improved using a linear potential sweep rather than a direct step to the target potential. Electrochemical impedance spectroscopy data relate information on the physical nature of the electrode/solution interface to the electrochemical performance of bare and enzyme-modified carbon-fiber microelectrodes. The electrodeposition waveform and scan rate were characterized for optimal membrane formation and performance. Finally, codetection of both DA/glucose and DA/lactate was demonstrated in intact rat striatum using probes fabricated according to the optimized protocol. Overall, this work improves the reliable fabrication of carbon-fiber microbiosensors for codetection of DA and important energetic substrates that are locally delivered to the recording site to meet metabolic demand.

The Oxidase Mimicking Activity of MnOx NPs/Co3O4 NPs Hybrid Nanozyme for Glucose Oxidation

Herein, the hybrid nanozyme MnOx NPs/Co3O4 NPs on indium tin oxide coated glass substrate (ITO) was manufactured by imparting the porous morphology with its distinct merits: its surface valence states, oxygen vacancies, large surface area, and abundant active sites. The oxidase-like activity was investigated via the catalytic oxidation of chromogenic substrate in the presence of glucose visualized by the eyes. MnOx NPs containing Mn2+ and Mn3+ have a superior ability to oxidize glucose by reducing dissolved oxygen and producing H2O2. Co3O4 NPs, in turn, reduce H2O2 with concomitant 3,3′,5,5′-tetramethylbenzidine (TMB) oxidization. Thus, the nanozyme mimics the dual roles of glucose oxidase and peroxidase. The oxidase-like activity of hybrid nanozyme for glucose was found to be higher than those of single components. The nanozyme responded to glucose with a linear range from 60 µM to 1200 μM. The acceptable performance is probably due to the facilitated access of glucose to the proximity of the sensor surface. Good reproducibility was accomplished by virtue of the meticulous construction of NPs. Without functionalization and enzyme utilization, the fabricated nanozyme holds promise as a substitute for peroxidase and oxidase for detecting glucose.

Improvement of Hydrogen Peroxide and Glucose Detection in Serum using Persistent Luminescent Nanoparticles: Impact of Synthesis Parameters and Particle Size

In this study, we report the preparation of a library of nanoparticles doped with chromium (ZGO) through two distinct syntheses conducted at 120°C for 12 hours or 24 hours, followed by a calcination at 500°C or 750°C. Using centrifugation, and starting from polydisperse samples, we demonstrated the ability to achieve different sizes in a monodisperse manner. Furthermore, our research revealed that ZGO react differently with hydrogen peroxide (H2O2) based on their size, synthesis duration, and calcination temperature. We observed the most significant amplification of persistent luminescence for particles prepared at 120°C for 12 hours and calcinated at 500°C, with a size of 100 nm, in the presence of 100 mM H2O2. This study enabled the detection of H2O2 in serum, with a detection limit of up to 2.1 µM. Additionally, we designed a glucose test for based on the responsiveness of ZGO PLNPs to H2O2. Using glucose oxidase, the glucose detection limit in serum was established around 0.13 µM, with a detection range between 0 ‐ 5 µM. These findings could open new avenues to further control the size, synthesis, calcination temperature, and persistent luminescence of PLNPs, thus enhancing their utility in the development of new in vitro biosensors.

A Portable Paper Strip Biosensor -based Smartphone for Detection of Glucose Content in Rice Samples

A paper strip biosensor was fabricated through the combination of horseradish peroxidase and glucose oxidase with 4-aminoantipyrine and phenol in order to produce a pink color. The color intensity was measured with a smartphone for quantitative analysis. To achieve highest paper strip biosensor performance, the concentrations of glucose oxidase, horseradish peroxidase, phenol, 4-aminoantypyrine, pH, and reaction time were studied. Under optimizations, the linear range of the glucose was 0.5-10 mM, and the detection limit was 0.17 mM. The detection of glucose was achieved within 7 min. The paper strip biosensor was successfully applied to the measurement of glucose in rice samples. The paper strip biosensor exhibits considerable promise as a portable instrument for the detection of glucose in rice and is also applicable to other types of samples. This research, nevertheless, necessitates controlled experimental conditions. Due to the fact that it impacts enzymatic activity.