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Despite decades of laboratory and clinical trials, breast cancer remains the main cause of cancer-related disease burden in women. Considering the metabolism destruction effect of metformin (Met) and cancer cell starvation induced by glucose oxidase (GOx), after their efficient delivery to tumor sites, GOx and Met may consume a large amount of glucose and produce sufficient hydrogen peroxide in situ. Herein, a pH-responsive epigallocatechin gallate (EGCG)-conjugated low-molecular-weight chitosan (LC-EGCG, LE) nanoparticle (Met–GOx/Fe@LE NPs) was constructed. The coordination between iron ions (Fe3+) and EGCG in this nanoplatform can enhance the efficacy of chemodynamic therapy via the Fenton reaction. Met–GOx/Fe@LE NPs allow GOx to retain its enzymatic activity while simultaneously improving its stability. Moreover, this pH-responsive nanoplatform presents controllable drug release behavior. An in vivo biodistribution study showed that the intracranial accumulation of GOx delivered by this nanoplatform was 3.6-fold higher than that of the free drug. The in vivo anticancer results indicated that this metabolism destruction/starvation/chemodynamic triple-combination therapy could induce increased apoptosis/death of tumor cells and reduce their proliferation. This triple-combination therapy approach is promising for efficient and targeted cancer treatment.

A novel Fe2+/Tyr/H2O2 fluorescence reaction system has been established for the purpose of analyzing glucose oxidase activity. This system involves the catalysis of glucose oxidase on glucose to produce H2O2, which in turn oxidizes tyrosine to a highly fluorescent substance under the catalysis of Fe2+. The fluorescence intensity is subsequently employed to ascertain the enzymatic activity of glucose oxidase. The enzymatic oxidation reaction and tyrosine fluorescence reaction conditions were optimized based on the H2O2 standard curve equation. Direct fluorescence spectrophotometry was used to determine the activity range and detection limit of glucose oxidase, which were found to be 7.00 × 10−5–7.00 × 10−2 U/mL and 3.36 × 10−5 U/mL (Enzyme-like activity is 6.72 × 10−4 U/mL, The enzyme reaction time is 5 min), respectively, with a relative standard deviation of less than 3.2 %. This method has been successfully applied to determine the activity of glucose oxidase in food additives, with a recovery rate ranging from 96.00 % to 102.0 %.

With the advancement of scientific research, an increasing number of studies on metal nanozymes, particularly Au NPs, have been conducted. They possess glucose oxidase (GOD)-like activity and peroxidase (POD)-like activity, with GOD-like activity receiving the most attention. However, there is a lack of research and summary on future trends in Au NPs in biomedical field. Therefore, we conducted a study on the GOD-like activity of Au NPs based on 158 relevant publications downloaded from the Web of Science Core Collection between 2007 and 2023, using citation metrics, authors, nations, institutions, cited references, and keywords. We found that Au NPs were initially utilized for glucose biosensors, glucose detection in human serum, and the detection of other carbohydrates and proteins. Researchers have recently focused on its application in oncotherapy and diabetic wound treatment. In addition to being utilized to treat autoimmune illnesses, inflammation, and other diseases, Au NPs may eventually replace insulin and antibiotics. We anticipate that the use of Au NPs will increase in the future. We hope this study will serve as a guide for future researchers so that Au NPs can be applied more broadly and competently.

Chemodynamic therapy (CDT) has been exploited as an emerging therapeutic modality for cancer treatment. However, insufficient intracellular H2O2 levels and ultrahigh glutathione (GSH) in the tumor microenvironment (TME) severely restrict CDT efficacy. To address these concerns, in this work, a self-reinforcing biodegradable nanotherapeutic was developed to modulate the TME to amplify CDT. The nanotherapeutic (GOD-FeCaP@TPZ, abbreviated GFCT) was fabricated through a mild one-step biomineralization process using the enzyme catalyst glucose oxidase (GOD) as a template to form biodegradable iron-doped calcium phosphate nanoparticles, followed by the loading of the prodrug tirapazamine (TPZ). When GFCT decomposes and releases Fe3+ in the acidic TME, the delivered GOD can effectively catalyze the oxidization of glucose to enhance the subsequent Fenton-like reaction by promoting intracellular acidity (via the production of gluconic acid) and improving the generation of H2O2. Moreover, the released iron constantly depletes GSH by self-cyclic valence alterations in Fe(III) and Fe(II) and then triggers H2O2 to produce •OH through an Fe(II)-mediated Fenton-like reaction. Moreover, the elevated level of hypoxia induced by GOD-mediated glucose oxidization can activate the prodrug TPZ for effective chemotherapy, thus achieving remarkable synergistic therapeutic outcomes. This work provides another paradigm to augment antitumor efficacy via modulation of the TME.

Glucose oxidase (GOx) can specifically catalyze the conversion of β-d-glucose into gluconic acid and hydrogen peroxide (H2O2) in the presence of oxygen, making it promising for tumor starvation therapy and oxidative therapy. However, GOx's immunogenicity, poor in vivo stability, short half-life, and potential systemic toxicity, limit its application in cancer therapy. Nanocarriers are capable of improving the pharmacological properties of therapeutic drugs (e.g. stability, circulating half-life, and tumor accumulation) and lower toxicity, hence resolving GOx issues and enhancing its efficacy. Although the application of targeted nanocarriers based on GOx has recently flourished, this field has not yet been reviewed and evaluated. Herein, we initially examined the mechanism of GOx-based nanocarriers for enhanced tumor therapy. Also, we present a comprehensive and up-to-date review that highlights GOx-based nanocarriers for tumor targeting therapy. This review expands on GOx-based nano-targeted combination therapies from both passive and active targeting perspectives, meanwhile, active targeting is further classified into ligand-mediated targeting and physical-mediated targeting. Furthermore, this review also emphasizes the present challenges and promising advancements.

Lactose concentration is a key parameter for assessing the quality of raw and processed milk and identifying abnormal milk. Current methods for lactose determination are laborious, time-consuming and oftentimes require skilled lab workers, expensive equipment, and special reagents. In this study, an optical biosensor system based on nanosized titanium dioxide (TiO2) is presented for routine measurement of lactose in milk without sample preparation. TiO2 was obtained via sol-gel synthesis and characterized using dynamic light scattering, microelectrophoresis, X-ray diffraction, transmission electron microscopy, and nitrogen adsorption-desorption analysis. The system, whose colorimetric response was based on the chromogen-free TiO2-based detection of hydrogen peroxide (H2O2), used β-galactosidase and/or glucose oxidase as the biorecognition elements. The selectivity and sensitivity of the biosensors for lactose and glucose was investigated by diffuse reflectance spectroscopy using standard aqueous lactose and glucose solutions and whole milk samples. The results showed that the proposed biosensors had high optical signal reproducibility (RSD = 5–6%), high sensitivity (LOD = 0.005 wt%), and high selectivity. The linear dynamic ranges of lactose and glucose detection were from 0.005 wt% to 0.100 wt%, and the optimal response time of the biosensor was 25 min. The listed features and advantages of label-free detection make the biosensor attractive for simple, inexpensive, sensitive, and selective determination of lactose in milk.

Since the discovery of acrylamide in heat-treated foods (crisps and biscuits) in April 2002, by a Swedish study, many efforts have been made to reduce acrylamide levels in foods. Consideration was given to reducing acrylamide levels by reducing Maillard reaction products. However, the production of potato chips creates acrylamide, a carcinogenic compound. This study aimed to use a new quantitative index and formula for l-asparaginase, glucose oxidase, their 1:1 blending enzymes, baker's yeast and green tea powder (0.5 g/kg distilled water) to reduce acrylamide production in potato chips and evaluate the products on some new indicators, such as asparagine reduction (%), acrylamide reduction (%), Asparagine / Acrylamide ratio (AAR) and Asparagine content (mg/kg). The highest acrylamide concentrations were reduced from 1346.0 ± 7.9 mg/kg in the blank sample (control) to 285.7 ± 3.2 and 325.3 ± 10.5 mg/kg in the baker's yeast sample (CT4) and green tea sample (CT5) treated sample, respectively. The potato chips treated with 0.5 g/kg l-asparaginase (CT1) reduced the acrylamide level by approximately 70.65±0.31%, while the CT4 samples showed 78.77±0.21% and asparagine contents of 116.0 ± 5.3 mg/kg and 195.0 ± 5.0 mg/kg, respectively.The results showed that some enzymes, baker's yeast and green tea powder could control the formation of acrylamide, which was formed during the processing of potato chips, allowing the food industry to produce low acrylamide foods from high starch food products such as potatoes. It has the ability to be used as a functional food, as well as to protect the human body of school-age children and adolescents from absorbing 70–80% of their daily acrylamide intake without these treatments.

Glucose oxidase (GOX) catalyzes the FAD-dependent oxidation of α-D-glucose to D-gluconolactone with production of hydrogen peroxide. This enzyme encounters many biotechnological applications from glucose sensors to applications in food, pharma and textile industries. For this purpose, recombinant GOX versions, usually derived from Aspergillus niger, are produced in fermentation systems, frequently in filamentous fungi because other production platforms such as bacteria or yeast have rendered meager results. We wondered whether A. niger GOX, more specifically a mutant version with superior thermotolerant properties, could be efficiently produced in Nicotiana benthamiana plants. To this aim, we used a tobacco mosaic virus-derived vector that is inoculated into plant tissues using Agrobacterium tumefaciens. Results exhibited the efficient production of the recombinant GOX in plants and the facile downstream purification when the recombinant protein is targeted to the apoplast, the space between plasma membranes and cell walls. The plant-made recombinant GOX displayed excellent catalytic properties in broad pH and temperature conditions. In addition to establishing a new strategy to produce recombinant GOX in plants as a green alternative to traditional fungal fermentation, we further investigated the potential application of this protein as an ezybiotic. Results exhibited a remarkable bacteriocide activity against Escherichia coli and Staphylococcus aureus.

Major challenges for cancer treatment are how to effectively eliminate primary tumor and sufficiently induce immunogenic cell death (ICD) to provoke a robust immune response for metastasis control. Here, a self-assembled cascade bioreactor was developed to improve cancer treatment with enhanced tumor penetration and synergistic therapy of starvation, chemodynamic (CDT) and photothermal therapy. Ultrasmall FeS-GOx nanodots were synthesized with glucose oxidase (GOx) as template and induced by paclitaxel (PTX) to form self-assembling FeS-GOx@PTX (FGP) via hydrophobic interaction. After accumulated at tumor sites, FGP disassembles to smaller FeS-GOx for enhanced deep tumor penetration. GOx maintains high enzymatic activity to catalyze glucose with assistant of oxygen to generate hydrogen peroxide (H2O2) as starvation therapy. Fenton reaction involving the regenerated H2O2 in turn produced more hydroxyl radicals for enhanced CDT. Following near-infrared laser at 808 nm, FGPs displayed pronounced tumor inhibition in vitro and in vivo by the combination therapy. The consequent increased exposure to calreticulin amplified ICD and promoted dendritic cells maturation. In combination with anti-CTLA4 checkpoint blockade, FGP can absolutely eliminate primary tumor and avidly inhibit distant tumors due to the enhanced intratumoral infiltration of cytotoxic T lymphocytes. Our work presents a promising strategy for primary tumor and metastasis inhibition.

Desugarization is an important technique in industrial egg processing to maintain the color and shelf life of egg white powder. Its effect on proteolysis of egg white protein is essential in understanding the functionality changes in the obtained hydrolysate. Two desugarization methods, including yeast fermentation and enzyme desugarization using glucose oxidase are commonly employed in industrial scale egg white processing but the suitable method which can improve egg white protein hydrolysates' functionality has not yet been reported. Thus, the present work compares the physicochemical, functional, and surface properties of egg white protein hydrolysates obtained through both the desugarization methods. Alcalase enzyme (2% w/w) was used for enzyme hydrolysis. The degree of hydrolysis of desugared egg white hydrolysate was higher than the non-desugared samples by 34.5%. The desugared samples (especially yeast fermented) had higher foaming and emulsifying ability than non-desugared samples; however, it showed emulsion activity index and surface hydrophobicity. Compared to native protein, the in-vitro trypsin digestibility of desugared hydrolysate samples was significantly (p

Intratumoral injections can reduce systemic absorption and deliver large amounts of drugs to the tumor, thereby reducing side effects and exhibiting high therapeutic efficacy. Therefore, a variety of drug delivery systems, such as hydrogels, fine particles, and nanoparticles, have been studied. Although the sustained-release drug delivery system can effectively reduce systemic absorption due to the slow release of the drug from the site of intratumoral injection, it lacks the ability to deliver high concentrations of drugs to the tumor. In particular, the larger the tumor size, the lower the efficacy of the treatment. To address this problem, this study focused on the tumor structure. Owing to the three-dimensional structure of the dense tumor microenvironment (TME) and abnormal blood vessels, drugs administered directly into the tumor act as if they were encapsulated in a hydrogel. To evaluate whether the three-dimensional structure of the tumor affects the intratumoral distribution and systemic absorption of drugs, needle-type starch implants (GOD-NS implants) and needle-type gelatin implants (GOD-NG implants) containing glucose oxidase (GOD), a protein that exhibits anti-cancer effects through hydrogen peroxide (H2O2) generation, were prepared. Both GOD-NS and GOD-NG implants can be easily injected into tumors. GOD-NS implants released GOD slowly, whereas GOD-NG implants released most of the GOD within 1 h. When a GOD-NG implant that rapidly released GOD was also injected, a high concentration of GOD was maintained in the tumor for a long time as it was trapped in the three-dimensional structure of the tumor. This study demonstrated that intratumoral injection of a rapidly drug-releasing gelatin needle may be a novel drug delivery system capable of long-term retention of high drug concentrations in tumors, as the three-dimensional structure of the tumor affects drug delivery.

The demand for glucose-sensing devices has increased along with the increasing diabetic population. Here, we aimed to construct a system with a glucose oxidase (GOx)-integrated Cu-nanoflower (Cu-NF) as the underlying electrode. This novel system was successfully developed by creating a cross-linked GOx within a Cu-NF matrix, forming a c-GOx@Cu-NF-coated film on a carbon screen-printed electrode (CSPE). A comparison of the stabilities of the cross-linking methods demonstrated enhanced durability, with an activity level of >88 % maintained after approximately 35 days of storage in room temperature buffer. Regarding the ability of the c-GOx@Cu-NF modified CSPE to detect glucose via electrochemical methods, the redox potential gap (ΔE) and peak current increased in the presence of GOx. In comparison to that of glucose, the sensitivity of c-GOx@Cu-NF was approximately 8 times greater than that of GOx@Cu-NF, with a detection limit of 0.649 μM and a linear range of 5-500 μM. It sustained an average relative activity of 80 % over 20 days. After 10 cycles of repeated use, the activity remained above 75 %. In terms of evaluating the electrode's specificity for glucose, the detection rate for individual similar substances was approximately 1 %. The introduction of a crosslinking strategy to Cu-NF, leading to enhanced mechanical stability and conductivity, improved the detection capability. Furthermore, this approach led to increased long-term storage stability and reusability, allowing for specific glucose detection. To our knowledge, this report represents the first demonstration of a c-GOx@Cu-NF system for integrating electrochemical biosensing devices into digital healthcare pathways, offering enhanced sensing accuracy and mechanical stability., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In recent years, the exceptional biocatalytic properties of glucose oxidase (GOx) have spurred the development of various GOx-functionalized nanocatalysts for cancer diagnosis and treatment. Carbon dots, renowned for their excellent biocompatibility and distinctive fluorescence properties, effectively incorporate GOx. Given the paramount importance of GOx's enzymatic activity in therapeutic efficacy, this study conducts a thorough exploration of the molecular-level binding dynamics between GOx and near-infrared carbon dots (NIR-CDs). Utilizing various spectrometric and molecular simulation techniques, we reveal that NIR-CDs form a ground-state complex with GOx primarily via hydrogen bonds and van der Waals forces, interacting directly with amino acid residues in GOx's active site. This binding leads to conformational change and reduces thermal stability of GOx, slightly inhibiting its enzymatic activity and demonstrating a competitive inhibition effect. In vitro experiments demonstrate that NIR-CDs attenuate the GOx's capacity to produce H 2 O 2 in HeLa cells, mitigating enzyme-induced cytotoxicity and cellular damage. This comprehensive elucidation of the intricate binding mechanisms between NIR-CDs and GOx provides critical insights for the design of NIR-CD-based nanotherapeutic platforms to augment cancer therapy. Such advancements lay the groundwork for innovative and efficacious cancer treatment strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this study, it was aimed to exploit no chemical drug to treat cancer by starvation-immunotherapy combination. We used PD-L1 siRNA to down-regulate checkpoint protein PD-L1 on cancer cell surfaces, and IFN-γ was used as an immunomodulatory agent to activate the innate immune system. When PD-L1 siRNA, IFN-γ, and glucose oxidase (GOx) were co-delivered to the tumor microenvironment (TME) by a pH-sensitive drug delivery system (DDS), starvation-immunotherapy combinedly resulted in significant tumor volume reduction in a mouse model. The DDS was prepared by using PLGA polymer whose surface was coated with chitosan. IFN-γ, siRNA, and GOx entrapment efficiencies were 93.5 ± 3.1%, 65.1 ± 2.6%, and 66.2 ± 3.3%, respectively. Moreover, the size of the DDS was well below 200 nm, allowing easy sterilization and tumor accumulation by the enhanced permeability and retention (EPR) effect. In vitro pH-dependent release study indicated that in acidic pH, the developed DDS can show accelerated drug release. In a mouse model and in acidic TME, the DDS resulted in the best therapeutic effect due to pH-dependent drug release compared to unmodified DDS and free drug. Our findings reflect that high efficiency in colon cancer treatment is achieved by combination therapy over the studied monotherapies.

Glucose oxidase (GOD), a popular enzyme used in pharmaceutical, food, bio-sensor and other fields, can well catalyze the oxidation of glucose to produce gluconic acid and hydrogen peroxide. The main limitation for the applications of free enzyme is its less preferable owing to its weaker activity against to changeable conditions such as pH, temperature, and organic solvent. The immobilization is thought to be an effective way to improve the stability of GOD under harsh conditions. In this study, MIL-125 was applied as a fixing carrier to anchor GOD for the first time. The synthesized MIL-125 was characterized by using X-ray diffraction (XRD), Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), N2 adsorption-desorption and DFT pore size distribution. The FT-IR spectra were employed to study the structural stability and interface interaction of MIL-125 with GOD. Compared to the previously reported GOD-related adsorption results, the higher adsorption capacity of GOD was obtained on MIL-125. The adsorption kinetics and isotherm analyses indicated that the adsorption process of GOD on MIL-125 was a spontaneous endothermic process, which was more in line with the pseudo-second-order kinetic model. The enzyme activity assay showed that the values of Km and Vmax for immobilized GOD on MIL-125 were 14.482 mmol L−1 and 0.356 mmol⋅L−1⋅min−1, respectively, which were lower than those of free GOD. But, compared to free GOD, the immobilized GOD showed less sensitivity to the changes in temperature, pH values and organic solvent. After six cyclic catalytic reactions, the immobilized GOD remained over 43.79% of its initial activity. The endurability tests showed that the immobilized GOD showed 43.41% of its initial activity after storage for 21 days at room temperature; meanwhile, the free GOD almost lost its initial activity at the end of 3 days under the same condition. The results demonstrated that the MOF-immobilized enzymes were more stable and had a broader potential application in the enzyme immobilization engineering.

The negative effects of dietary antibiotics have become a widespread concern. It is imperative to search for a new type of green, safe, and efficient feed additive that can replace antibiotics. This study was to investigate the effects of glucose oxidase (GOD) on growth performance, immune function, and intestinal barrier in ducks infected with Escherichia coli O88. First, we established the E. coli challenge model of ducks through a preliminary experiment and then carried out the formal experiment by using 144 1-day-old male lean Peking ducklings (50 ± 2.75 g). All ducks were randomly assigned to 1 of 3 dietary treatment groups of basal diet (control), 30 mg/kg virginiamycin (antibiotic), and 200 U/kg GOD (1,000 U/g). Each group consisted of 6 replications with 8 birds per replicate. At day 7, all ducks were orally administered 0.2 mL E coli O88 (3 × 109 cfu/mL) twice, 8 h apart based on the preliminary experiment. The experiment lasted for 28 d. Dietary supplementation with GOD improved growth performance of ducks infected with E. coli. The GOD increased contents of Ig in plasma and secreted Ig A in jejunal mucosa. The GOD group had lower concentrations of inflammatory cytokines (tumor necrosis factor-α, IL-1β, and IL-6) and their upstream regulator Toll-like receptor 4 in the jejunum of ducks than the control group. Supplementation with GOD increased villus height and decreased crypt depth in the jejunum. The gene expression of tight junction proteins (zonula occludens-1, claudin-1 and claudin-2) was enhanced by adding GOD. The GOD decreased intestinal permeability by reducing the concentrations of diamine oxidase and D-lactic in plasma of ducks. There were no significant differences in almost all the indices tested between the GOD and the antibiotic groups. In conclusion, supplementation of GOD improved growth performance, immune function, and intestinal barrier of ducks infected with E. coli O88. Glucose oxidase may serve as a promising alternative therapy to antibiotics to relieve or prevent colibacillosis in ducks.

Herein, a comparative study between novel water-soluble phthalocyanine-based biosensors was performed for the application of glucose sensing. For this purpose, two different copper (II) and manganese (III) phthalocyanines and their water-soluble derivatives were synthesized, and then their role as a supporting material for enzyme immobilization was evaluated by comparing their sensor performances. Two different phthalocyanine (AP-OH 2 -MnQ (MnPc) and AP-OH 2 -CuQ (CuPc)) were tested using electrochemical biosensor with immobilized glucose oxidase (GOx). To the best of our knowledge, the related water-soluble phthalocyanine-based glucose biosensors were attempted for the first time, and the developed approach resulted in improved biosensor characteristics. The constructed biosensors GE/MnPc/GOx and GE/CuPc/GOx showed good linearity between 0.003-1.0 mM and 0.05-0.4 mM, respectively. The limit of detection was estimated at 0.0026 mM for the GE/MnPc/GOx and 0.019 mM for the GE/CuPc/GOx. K M app and sensitivity values were also calculated as 0.026 mM and 175.043 µAmM -1 cm -2 for the GE/MnPc/GOx biosensor and 0.178 mM and 117.478 µAmM -1 cm -2 for the GE/CuPc/GOx biosensor. Moreover, the fabricated biosensors were successfully tested to detect glucose levels in beverages with high recovery results. The present study shows that the proposed water-soluble phthalocyanines could be a good alternative for quick and cheap glucose sensing with improved analytical characteristics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

ABSTRACT: Arbor Acre (AA) broilers were used as the research object to investigate whether glucose oxidase (GOD) has preventive and relieving effects on necrotic enteritis. The experiment was designed as a factorial arrangement of 2 dietary treatments × 2 infection states. Chickens were fed a basal diet or a diet with 150 U/kg GOD, and were challenged with Clostridium perfringens (Cp) or sterile culture medium. In our study, Cp challenge led to intestinal injury, as evidenced by reducing the average daily gain and the average daily feed intake of AA broilers of 14 to 21 d (P < 0.05), increasing the intestinal jejunal lesion score (P < 0.05), reducing the jejunal villi height and villi height/crypt depth (P < 0.05), upregulating the mRNA expression levels jejunal IFN-γ (P < 0.05). The dietary GOD had no significant effects on the growth performance of each growth period, but significantly decreased the ileal pH, increased the height of villi and the ratio of villi height to crypt depth (P < 0.05) and the expression levels of Occludin and Zonula occludens-1 (ZO-1) at d 21. Moreover, dietary GOD and the Cp challenge significantly altered the composition of 21-d ileal microbiota. The Cp challenge decreased the relative abundance of genus Lactobacillus (P = 0.057), and increased the relative abundance of genus Romboutsia (P < 0.05) and genus Veillonella (P = 0.088). The dietary GOD tended to increase the relative abundance of genus Helicobacter (P = 0.066) and decrease the relative abundance of genus Streptococcus (P = 0.071). This study has shown that the supplementation of GOD could promote the integrity of intestinal barrier and the balance of ileal microbiota, but the effects of GOD on NE broilers and its application in actual production need to be further confirmed.

A surface plasmon resonance imaging (SPRI)-based biosensor is demonstrated for the detection of both hydrogen peroxide (H 2 O 2 ) and glucose. The H 2 O 2 to be detected acts as an oxidant and etch the silver film. This process gradually effects on resonance condition and consequently the reflected light intensity at a fixed angle. The etching rate of the silver film shows a clear relation with the H 2 O 2 concentration. Therefore, monitoring the reflected light intensity progressively changing over a few minutes, enables accurate detection of H 2 O 2 concentrations ranging from 0 to 200 μM (within physiological range of 0.25-50 μM), with a remarkable limit of detection (LOD) as low as 40 nM. In this regard, the behavior of the surface plasmon resonance (SPR) dip in response to the reduction of the silver film thickness is predicted by Winspall simulation software. These simulation results are in good agreement with the experimental results. Moreover, the proposed method can be applied to determine glucose concentrations ranging from 0 to 10 mM, encompassing the physiological range of 3-8 mM. This is achieved by observing the generated H 2 O 2 through the enzymatic oxidation reaction between glucose and glucose oxidase (Gox). The sensor demonstrates remarkable sensitivity and selectivity, with a detection limit as low as 175 μM for glucose concentration. Furthermore, accurate measurement of glucose concentration in an actual human serum sample is achievable with the proposed sensor, using the standard addition method. The suggested glucose sensor shows promising prospects for use in routine glucose testing, employing a label-free, real-time, and multiplex detection approach.© 2017 Elsevier Inc. All rights reserved., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The recognition of silver nanoparticles (AgNPs) as a nanozyme with peroxidase-like activity has offered a promising solution to address the challenges of bacterial resistance and argyria risk. However, the catalytic efficacy of AgNPs is limited by the need for a strong acidic environment and high concentrations of hydrogen peroxide (H 2 O 2 ). In this work, we developed a self-activated hydrogel cascade reactor (AUGP) for enhanced treatment of bacterial infection. The AUGP integrates the properties of glucose oxidase (GOx) and polyacrylamide (pAAm) hydrogel microsphere. The confinement effect of pAAm hydrogel microsphere enables glucose oxidation to occur in a confined space, which creates an acidic environment to activate AgNPs activity, initiating the cascade reaction between GOx and AgNPs. Meanwhile, the confinement effect facilitates the accumulation of a high local concentration of H 2 O 2 , allowing AUGP to generate hydroxyl radicals (•OH) without the need for external H 2 O 2 . Additionally, the release of Ag + from AUGP is achieved upon the generation of •OH. The synergistic action of Ag + and •OH confers exceptional antibacterial efficacy to AUGP. Importantly, the etching effect of H 2 O 2 ensures the absence of any residual AgNPs, reducing the risk of argyria. In vivo studies validated the efficacy of AUGP in wound disinfection with minimal toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Glucose oxidase (GOx) is often used to starvation therapy. However, only consuming glucose cannot completely block the energy metabolism of tumor cells. Lactate can support tumor cell survival in the absence of glucose. Here, we constructed a nanoplatform (Met@HMnO 2 -GOx/HA) that can deplete glucose while inhibiting the compensatory use of lactate by cells to enhance the effect of tumor starvation therapy. GOx can catalyze glucose into gluconic acid and H 2 O 2 , and then HMnO 2 catalyzes H 2 O 2 into O 2 to compensate for the oxygen consumed by GOx, allowing the reaction to proceed sustainably. Furthermore, metformin (Met) can inhibit the conversion of lactate to pyruvate in a redox-dependent manner and reduce the utilization of lactate by tumor cells. Met@HMnO 2 -GOx/HA nanoparticles maximize the efficacy of tumor starvation therapy by simultaneously inhibiting cellular utilization of two carbon sources. Therefore, this platform is expected to provide new strategies for tumor treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

The development of rapid screening sensing platforms to improve pre-screening mechanisms in community healthcare is necessary to meet the significant need for portable testing in biomarker diagnostics. Here, we designed a portable smartphone-based photoelectrochemical (PEC) immunoassay for carcinoembryonic antigen (CEA) detection using Cu-doped ultrathin porous Bi 2 WO 6 (CuBWO) nanosheets as the photoactive material. The CuBWO nanosheets exhibit a fast photocurrent response and excellent electrical transmission rate under UV light due to their surface plasmon resonance effect (SPR). The method uses glucose oxidase-labeled secondary antibody as a signal indicator for sandwich-type immune conjugation. In the presence of the target CEA, the electrons and holes generated at the surface of the photo-excited ultrathin porous CuBWO were rapidly consumed by the production of H 2 O 2 from glucose oxidase oxidizing glucose, resulting in a weakened photocurrent signal. The photocurrent intensity increased logarithmically and linearly with increasing CEA concentration (0.02-50 ng mL -1 ), with a detection limit of 15.0 pg mL -1 (S/N = 3). The system provides a broader idea for inferring the electron-hole transport mechanism in ultrathin porous nanosheet layer materials and developing efficient PEC sensors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The oxygen evolution reaction (OER) is an important half-reaction in electrochemical hydrogen production (EHP) and rechargeable metal-air batteries. However, the sluggish OER kinetics has seriously impeded their performance. Herein, we report a bioelectrochemical cascade system composed of glucose oxidase (GOx)-functionalized N-doped porous carbon nanofibers to replace OER in EHP and rechargeable Zn-air batteries (ZABs) applications. In this cascade system, GOx catalyzes oxidation of glucose to produce value-added gluconic acid accompanied with the generation of H 2 O 2 under aerobic conditions. The subsequent electrocatalytic oxidation of H 2 O 2 replacing the OER results in an onset voltage below 1.10 V for EHP, and a low charging voltage of 1.35 V as well as a small charging/discharging voltage gap of ∼ 280 mV over 170 h for ZABs in neutral aqueous electrolytes. The advantages of employing the innovative bioelectrochemical cascade reaction are demonstrated in EHP and ZABs, achieving the full utilization of biomass energy in energy-saving electrochemical systems for energy storage and conversion., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study introduces the UV/glucose-oxidase@Kaolin (GOD@Kaolin) coupled organic green rust (OGR) system (UV/OGR/GOD@Kaolin) to investigate the promotion of glucose oxidase activity by UV light and its synergistic degradation mechanism for photosensitive pollutants, specifically targeting the efficient degradation of 4-chlorophenol (4-CP). The enzyme system demonstrates its ability to overcome drawbacks associated with traditional Fenton systems, including a narrow pH range and high localized concentration of H 2 O 2 , by gradually releasing hydrogen peroxide in situ within a neutral environment. In the presence of UV radiation under specific conditions, enhanced enzyme activity is observed, resulting in increased efficiency in pollutant removal. The gradual release of hydrogen peroxide plays a crucial role in preventing unwanted reactions among active substances. These unique features facilitate the generation of highly reactive species, such as Fe(IV)O, •OH, and •O 2 - , tailored to efficiently target the organic components of interest. Additionally, the system establishes a positive iron cycle, ensuring a sustained reactive capability throughout the degradation process. The results highlight the UV/OGR/GOD@Kaolin system as an effective and environmentally friendly approach for the degradation of 4-CP, and the resilience of the enzyme extends the system's applicability to a broader range of scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Fluorescent copper nanoparticles (CuNPs) was synthesized by one-step chemical reduction method using ascorbic acid (AA) and copper sulfate (CuSO 4 ⋅5H 2 O) as raw materials, which had good water solubility and fluorescence properties. A green, simple and safe CuNPs@Fe 2+ fluorescence probe was developed for the detection of hydrogen peroxide and glucose using Fe 2+ as a bridge. The prepared CuNPs could obtain the maximum fluorescence emission wavelength at 440 nm when the excitation wavelength was 360 nm. The average particle size of CuNPs was 10 nm, which had good photobleach resistance, stability and salt tolerance. The fluorescence intensity was quenched due to electron transfer (ET) process when hydrogen peroxide was added to CuNPs@Fe 2+ system. This result was mainly because Fenton reaction occured between hydrogen peroxide and Fe 2+ , producing hydroxyl free radicals (OH) and Fe 3+ . Since glucose could be catalyzed by specific glucose oxidase (GOX) to produce H 2 O 2 and corresponding oxidation products, the quantitative analysis of glucose was realized when glucose oxidase was introduced into the CuNPs@Fe 2+ sensor system. Therefore, a novel CuNPs@Fe 2+ fluorescent probe sensor study was constructed to further achieve quantitative detection of H 2 O 2 and glucose. Under the optimized experimental conditions, the linear ranges for H 2 O 2 and glucose were 28.219-171.562 μM and 1.237-75.771 μM, respectively. And the detection limits for H 2 O 2 and glucose were 7.169 μM and 0.540 μM, respectively. In addition, the mechanism of fluorescence probe quenching caused by the interaction between H 2 O 2 and CuNPs@Fe 2+ was also discussed. The proposed sensing system had been applied successfully to the detection of glucose in human serum samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: In vitro screening strategies based on the inhibition of α-glucosidase (GAA) activity have been widely used for the discovery of potential antidiabetic drugs, but they still face some challenges, such as poor enzyme stability, non-reusability and narrow range of applicability. To overcome these limitations, an in vitro screening method based on GAA@GOx@Cu-MOF reactor was developed in our previous study. However, the method was still not satisfactory enough in terms of construction cost, pH stability, organic solvent resistance and reusability. Thence, there is still a great need for the development of in vitro screening methods with lower cost and wider applicability., Results: A colorimetric sensing strategy based on GAA/(Au-Au/IrO 2 )@Cu(PABA) cascade catalytic reactor, which constructed through simultaneous encapsulating Au-Au/IrO 2 nanozyme with glucose oxidase-mimicking and peroxidase-mimicking activities and GAA in Cu(PABA) carrier with peroxidase-mimicking activity, was innovatively developed for in vitro screening of GAA inhibitors in this work. It was found that the reactor not only exhibited excellent thermal stability, pH stability, organic solvent resistance, room temperature storage stability, and reusability, but also possessed cascade catalytic performance, with approximately 12.36-fold increased catalytic activity compared to the free system (GAA + Au-Au/IrO 2 ). Moreover, the in vitro GAA inhibitors screening method based on this reactor demonstrated considerable anti-interference performance and detection sensitivity, with a detection limit of 4.79 nM for acarbose. Meanwhile, the method owned good reliability and accuracy, and has been successfully applied to the in vitro screening of oleanolic acid derivatives as potential GAA inhibitors., Significance: This method not only more effectively solved the shortcomings of poor stability, narrow scope of application, and non-reusability of natural enzymes in the classical method compared with our previous work, but also broaden the application scope of Au-Au/IrO 2 nanozyme with glucose oxidase and peroxidase mimicking activities, and Cu(PABA) carrier with peroxidase mimicking activity, which was expected to be a new generation candidate method for GAA inhibitor screening., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Hexaric acids have attracted attention lately because they are platform chemicals for synthesizing pharmaceuticals. In particular, gluconic acid is one of the most studied because it is readily available in nature. In this work, operational conditions like temperature and pH were evaluated for the enzymatic production of gluconic acid. For this purpose, glucose oxidase (GOx) and catalase (CAT) were individually immobilized and co-immobilized using amino-silica as support. The catalytic performance of the enzymes both as separate biocatalysts (GOx or CAT) and as an enzymatic complex (GOx-CAT) was assessed in terms of enzymatic activity and stability at temperatures 45 °C and 50 °C and pH 6 to 8. The results show that CAT is a key enzyme for gluconic acid production as it prevents GOx from being inhibited by H 2 O 2 . However, CAT was found to be less stable than GOx. Therefore, different GOx to CAT enzymatic ratios were studied, and a ratio of 1-3 was determined to be the best. The highest glucose conversion conditions were 45 °C and pH 7.0 for 24 h. Regarding the biocatalyst reuse, GOx-CAT retained more than 70% of its activity after 6 reaction cycles. These results contribute to further knowledge and application of oxidases for hexaric acid production and shed greater light on the role of the glucose oxidase/catalase pair in better catalytic performance. Both enzymes were immobilized in one pot, which is relevant for their potential use in industry; an enzyme system was obtained in a single step., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Enzymatic electrochemical biosensor is the most common analytical platform for medical diagnosis. To mimic the biological environment of the enzyme for maintaining the function of biosensor, zwitterionic hydrogels have been recognized as effective matrices for enzymatic immobilization. Herein, a zwitterionic hydrogel derived from a copolymer, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-N-methacryloyloxyethyl tyrosine methylester (MAT)] (PMM) was firstly applied as versatile coating to preserve stability and activity of oxidase enzymes, glucose oxidase (GOx) and lactate oxidase (LOx) for enzymatic electrochemical sensor. A screen-printed carbon electrode (SPCE) was sequentially coated with nitrogen-doped graphene (NDG), oxidase enzyme, and PMM mixed with Ru(II)bpy 3 2+ and (NH 4 ) 2 S 2 O 8 followed by visible light irradiation for 3 min to induce PMM gelation. Electrochemical detection of glucose and lactate using the modified SPCE was performed via amperometry in the presence of hydrogen peroxide. The activity of both GOx and LOx immobilized on the modified SPCE was well maintained for 49 days at 87 and 80 %, respectively. Additionally, two different electrodes, a screen-printed graphene electrode (SPGE), and a screen-printed silver electrode (SPAgE), similarly modified gave the same satisfactory detection of spiked glucose and lactate in human plasma and sweat with 93-118 % recovery. This indicates the potential of the PMM hydrogel as a universal platform for preservation of enzymes which can be easily fabricated without the need for specific chemical modification of the electrode., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The development of a self-regulated minimally invasive system for insulin delivery can be considered as the holy grail in the field of diabetes mellitus. A delivery system capable of releasing insulin in response to blood glucose levels would significantly improve the quality of life of diabetic patients, eliminating the need for frequent finger-prick tests and providing better glycaemic control with lower risk of hypoglycaemia. In this context, the latest advances in glucose-responsive microneedle-based transdermal insulin delivery are here compiled with a thorough analysis of the delivery mechanisms and challenges lying ahead in their clinical translation. Two main groups of microneedle-based systems have been developed so far: glucose oxidase-containing and phenylboronic acid-containing systems. Both strategies in combination have also been tested and two other novel strategies are under development, namely electronic closed-loop and glucose transporter-based systems. Results from preclinical studies conducted using these different types of glucose-triggered release systems are comprehensively discussed. Altogether, this analysis from both a mechanistic and translational perspective will provide rationale and/or guidance for future trends in the research hotspot of glucose-responsive microneedle-based insulin delivery systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Single-cell nanoencapsulation (SCNE) is a nanoarchitectonic strategy for creating cell-in-shell structures, in which the artificial shells, formed on individual living cells, protect the cells inside from otherwise lethal factors and also potentially provide them with advanced functions, such as exogenous biochemical reactions that are not attainable in wild-type cells. This work investigated enzymatic cascade systems for widening the substrate scope, beyond catecholamines, in the in-vitro formation of melanin-like films and shells and, ultimately, providing advanced building blocks and tools to the field of SCNE, inspired by the enzyme-derived structural diversification of melanin found in nature. The combination of glucose oxidase and horseradish peroxidase enabled the facilitated film formation of amine group-absent ortho-diphenols, such as protocatechuic aldehyde (PCA) and pyrocatechol. As a proof-of-demonstration, the developed reaction protocol was applied to the cytocompatible SCNE of Saccharomyces cerevisiae.

We investigated the effects of housing systems and dietary glucose oxidase (GOD) on the growth performance and intestinal health of Beijing You chickens (BYC). The experiment was designed as a factorial arrangement of 2 housing systems × 2 dietary treatments. Chickens were fed a basal diet or a diet with 200 U/kg GOD and were reared on the floor with deep litter or in the cages. Compared with the litter floor groups, the decreased average daily feed intake of 1 to 42 d, decreased feed conversion ratio (FCR), improved average daily gain of 42 to 77 d, and the whole period were identified in the cage rearing groups (P

Mechanical pre-treatment (disc refining) of wheat straw, at both atmospheric and elevated pressure, is shown to be an efficient process to access fermentable monosaccharides, with the potential to integrate within the infrastructure of existing first-generation bioethanol plants. The mild, enzymatic degradation of this sustainable lignocellulosic biomass affords ca. 0.10–0.13 g/g (dry weight) of d-glucose quantifiable voltammetrically in real time, over a two hundred-fold range in experimental laboratory scales (25 mL to 5.0 L), with pressure disc refining of the wheat straw enabling almost twice the amount of d-glucose to be generated during the hydrolysis stage than experiments using atmospheric refining (0.06–0.09 g/g dry weight). Fermentation of the resulting hydrolysate affords 0.08–0.10 g/g (dry weight) of ethanol over similar scales, with ethanol productivity at ca. 37 mg/(L h). These results demonstrate that minimal cellulose decomposition occurs during pressure refining of wheat straw, in contrast to hemicellulose, and suggest that the development of green, mechanochemical processes for the scalable and cost-effective manufacture of second-generation bioethanol requires improved cellulose decomposition.

The noble metal NPs that are currently applied to photothermal therapy (PTT) have their photoexcitation location mainly in the NIR-I range, and the low tissue penetration limits their therapeutic effect. The complexity of the tumor microenvironment (TME) makes it difficult to inhibit tumor growth completely with a single therapy. Although TME has a high level of H 2 O 2 , the intratumor H 2 O 2 content is still insufficient to catalyze the generation of sufficient hydroxide radicals (‧OH) to achieve satisfactory therapeutic effects. The AuPd-GOx-HA (APGH) was obtained from AuPd bimetallic nanodumbbells modified by glucose oxidase (GOx) and hyaluronic acid (HA) for photothermal enhancement of tumor starvation and cascade catalytic therapy in the NIR-II region. The CAT-like activity of AuPd alleviates tumor hypoxia by catalyzing the decomposition of H 2 O 2 into O 2 . The GOx-mediated intratumoral glucose oxidation on the one hand can block the supply of energy and nutrients essential for tumor growth, leading to tumor starvation. On the other hand, the generated H 2 O 2 can continuously supply local O 2 , which also exacerbates glucose depletion. The peroxidase-like activity of bimetallic AuPd can catalyze the production of toxic ‧OH radicals from H 2 O 2 , enabling cascade catalytic therapy. In addition, the high photothermal conversion efficiency (η = 50.7 %) of APGH nanosystems offers the possibility of photothermal imaging-guided photothermal therapy. The results of cell and animal experiments verified that APGH has good biosafety, tumor targeting, and anticancer effects, and is a precious metal nanotherapeutic system integrating glucose starvation therapy, nano enzyme cascade catalytic therapy, and PTT therapy. This study provides a strategy for photothermal-cascade catalytic synergistic therapy combining both exogenous and endogenous processes. STATEMENT OF SIGNIFICANCE: AuPd-GOx-HA cascade nanoenzymes were prepared as a potent cascade catalytic therapeutic agent, which enhanced glucose depletion, exacerbated tumor starvation and promoted cancer cell apoptosis by increasing ROS production through APGH-like POD activity. The designed system has promising photothermal conversion ability in the NIR-II region, simultaneously realizing photothermal-enhanced catalysis, PTT, and catalysis/PTT synergistic therapy both in vitro and in vivo. The present work provides an approach for designing and developing catalytic-photothermal therapies based on bimetallic nanoenzymatic cascades., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Tumor recurrence and wound healing represent significant burdens for tumor patients after the surgical removal of melanomas. Wound dressings with wound healing and anticancer therapeutic abilities could help to solve these issues. Thus, a hybrid hydrogel made of polyvinyl alcohol (PVA) and polyethylene imine (PEI) was prepared by cross-linking imine bond and boronic acid bond. This hydrogel was loaded with ruthenium nanorods (Ru NRs) and glucose oxidase (GOx) and named as nanocomposite hydrogel (Ru/GOx@Hydrogel), exhibiting remarkable photothermal/photodynamic/starvation antitumor therapy and wound repair abilities. Ru NRs are bifunctional phototherapeutic agents that simultaneously exhibit intrinsic photothermal and photodynamic functions. Three-dimensional composite hydrogel loaded with GOx can also consume glucose in the presence of O 2 during tumor starvation therapy. Near-infrared (NIR) light-triggered hyperthermia can not only promote the consumption of glucose, but also facilitate the ablation of residual cancer cells. The antitumor effect of the Ru/GOx@Hydrogel resulted in significant improvements, compared to those observed with either phototherapy or starvation therapy alone. Additionally, the postoperative wound was substantially healed after treatment with Ru/GOx@Hydrogel and NIR irradiation. Therefore, the Ru/GOx@Hydrogel can be used as a multi-stimulus-responsive nanoplatform that could facilitate on-demand controlled drug release, and be used as a promising postoperative adjuvant in combination therapy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Maillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed., Results: Herein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H 2 O 2 and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01-100 μM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations., Significance: This colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Glucose oxidase (GOD) is widely used in the pharmaceutical industry, fermentation products and glucose biosensors for its essential role in catalyzing the conversion of glucose to gluconic acid and hydrogen peroxide (H 2 O 2 ). As H 2 O 2 is the by-product and will have a toxic effect on glucose oxidase, so introducing another enzyme that could consume H 2 O 2 to form an enzymatic cascade reaction is a practical solution. However, this decision will lead to extra expenses and complex condition optimization such as the specific mass ratio, temperature and pH to improve the activity, stability and recyclability. Herein, we describe a mild and versatile strategy by anchoring GOD on carboxyl-activated MOF (Cu-TCPP(Fe)) through DNA-directed immobilization (DDI) technology. Robust MOF nanosheets were utilized as not only the carrier for the immobilization of GOD, but also a peroxidase-like catalyst for the decomposition of H 2 O 2 to reduce its harmful impacts. In this work, the immobilized GOD retained 55.78% of its initial activity after being used for 7 times. More than 60% of the immobilized enzyme's catalytic activity was still maintained after 96 h of being stored at 50 ℃. This study provides a new idea for preparing immobilized enzymes with enhanced stability, fast diffusion and high activity, which can be used in fields such as biocatalysis and biotechnology., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

Wearable microneedle sensors for continuous glucose monitoring (CGM) have great potential for clinical impact by allowing access to large data sets to provide individualized treatment plans. To date, their development has been challenged by the accurate wide linear range tracking of interstitial fluid (ISF) glucose (Glu) levels. Here, we present a CGM platform consisting of a three-electrode microneedle electrochemical biosensor and a fully integrated radio-chemical analysis system. The long-term performance of the robust CGM on diabetic rats was achieved by electrodepositing Prussian blue (PB), and crosslinking glucose oxidase (GOx) and chitosan to form a 3D network using glutaraldehyde (GA). After redox by GOx, PB rapidly decomposes hydrogen peroxide and mediates charge transfer, while the 3D network and graphite powder provide enrichment and release sites for Glu and catalytic products, enabling a sensing range of 0.25-35 mM. Microneedle CGM has high sensitivity, good stability, and anti-interference ability. In diabetic rats, CGM can accurately monitor Glu levels in the ISF in real-time, which are highly consistent with levels measured by commercial Glu meters. These results indicate the feasibility and application prospects of the PB-based CGM for the clinical management of diabetes. STATEMENT OF SIGNIFICANCE: This study addresses the challenge of continuous glucose monitoring system design where the narrow linear range of sensing due to the miniaturization of sensors fails to meet the monitoring needs of clinical diabetic patients. This was achieved by utilizing a three-dimensional network of glutaraldehyde cross-linked glucose oxidase and chitosan. The unique topology of the 3D network provides a large number of sites for glucose enrichment and anchors the enzyme to the sensing medium and the conductive substrate through covalent bonding, successfully blocking the escape of the enzyme and the sensing medium and shortening the electron transfer and transmission path., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Xuefeng Hu reports financial support was provided by National Key Research and Development Program of China, Major Science and Technology Project of Sichuan Province. Jieyu Zhang reports financial support was provided by Key Research and Development Program of Sichuan Province., (Copyright © 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Wearable biosensors have gained huge interest due to their potential for real-time physiological information. The development of a non-invasive blood glucose device is of great interests for health monitoring in reducing the diabetes incidence. Here, we report a sandwich-structured biosensor that is designed for glucose levels detection by using sweat as the means of monitoring. The Prussian blue nanoparticles (PBNPs) and carboxylated carbon nanotubes (MWCNT-COOH) were self-assembled on the electrode to improve the electrochemical performance and as the sensor unit, glucose oxidase (GOx) was immobilized by chitosan (CS) as the reaction catalysis unit, and finally encapsulated with Nafion to ensure a stable performance. As a result, the GOx/PBNPs/MWCNT-COOH sensor displays a low detection limit (7.0 μM), high sensitivity (11.87 μA mM -1  cm -2 ), and excellent interference resistance for a full sweat glucose application range (0.0-1.0 mM) for both healthy individuals and diabetic patients. Additionally, the glucose sensor exhibits stable stability for two weeks and can be successfully applied to screen-printed carbon electrodes (SPCE), demonstrating its great potential for personalized medical detection and chronic disease management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Fluorescence immunoassays are commonly employed for the detection of pathogenic bacteria as a means of ensuring food safety and preserving public health. However, the challenges such as poor photostability and background interference have limited their sensitivity and accuracy. The emergence of metal-organic frameworks (MOFs) as a label probe offers a promising solution for advancing fluorescence immunoassays owing to their tunable nature. Nonetheless, the low fluorescence efficiency of MOFs and the potential risk of dye leakage pose obstacles to achieving high detection sensitivity. Therefore, there exists a pressing need to fully utilize the potential of MOF composites in fluorescence immunoassays., Results: We explored the potential of glucose oxidase-encapsulated zeolitic imidazole framework-90 (GOx@ZIF-90) as a label probe to construct a time-resolved fluorescence immunoassay with amplified detection signal. This immunoassay involved functionalizing Fe 3 O 4 nanoparticle with porcine antibody to specifically capture and separate the target bacteria, Staphylococcus aureus (S. aureus). The captured S. aureus was then bound by GOx@ZIF-90 modified with vancomycin, resulting in a fluorescence response in the europium tetracycline (EuTc). The encapsulation of GOx in ZIF-90 provided a confinement effect that significantly enhanced the catalytic activity and stability of GOx. This led to a highly efficient conversion of glucose to H 2 O 2 , amplifying the fluorescence signal of EuTc. The immunoassay demonstrated a high sensitivity in detecting S. aureus, with a detection limit of 2 CFU/mL. We also obtained satisfactory results in milk samples. Attractively, the time-resolved detection mode of EuTc allowed the immunoassay to eliminate background fluorescence and enhance accuracy., Significance: This study not only presented a new method for detecting foodborne pathogens but also highlighted the potential of enzyme-encapsulated MOF composites as label probes in immunoassays, providing valuable insights for the design and fabrication of MOF composites for various applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The substrates of oxidase are biologically essential substances that are closely associated with human physiological health. However, current biosensing methods suffer from tough recyclability and undesired denaturation of enzyme due to impurity interference. Herein, we have developed a visual and reusable biosensor for detecting substrate using glucose oxidase (GOx) as a model oxidase. GOx was immobilized onto gold nanoparticles (AuNPs) at -20 °C in one step without additional reagents. The resulting nano-enzyme generated coloimetric signals by coupling with horseradish peroxidase (HRP) using TMB as the substrate. Our results demonstrated that the immobilized GOx exhibited satisfactory sensitivity (0.68 μM) for glucose detection and higher inherent stability than free GOx under harsh conditions, enabling reliable detection of glucose in complex fluids (colored beverages and saliva). Furthermore, the nano-enzyme retained 80 % activity even after four cycles of catalytic oxidation. This strategy constructs a universal biosensor for substrates with nano-enzyme which rely only on intrinsic cysteine within the oxidase while avoiding functional handle modification., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Magnetic nanoparticles (MNPs) are promising in tumor treatments due to their capacity for magnetic hyperthermia therapy (MHT), chemodynamic therapy (CDT), and immuno-related therapies, but still suffer from unsatisfactory tumor inhibition in the clinic. Insufficient hydrogen peroxide supply, glutathione-induced resistance, and high-density extracellular matrix (ECM) are the barriers. Herein, we hierarchically decorated MNPs with disulfide bonds (S-S), dendritic L-arginine (R), and glucose oxidase (GOx) to form a nanosystem (MNPs-SS-R-GOx). Its outer GOx layer not only enhanced the H 2 O 2 supply to produce . OH by Fenton reaction, but also generated stronger oxidants (ONOO - ) together with the interfaced R layer. The inner S-S layer consumed glutathione to interdict its reaction with oxidants, thus enhancing CDT effects. Importantly, the generated ONOO - tripled the MMP-9 expression to induce ECM degradation, enabling much deeper penetration of MNPs and benefiting CDT, MHT, and immunotherapy. Finally, the MNPs-SS-R-GOx demonstrated a remarkable 91.7% tumor inhibition in vivo. STATEMENT OF SIGNIFICANCE: Magnetic nanoparticles (MNPs) are a promising tumor therapeutic agent but with limited effectiveness. Our hierarchical MNP design features disulfide bonds (S-S), dendritic L-arginine (R), and glucose oxidase (GOx), which boosts H 2 O 2 supply for ·OH generation in Fenton reactions, produces potent ONOO - , and enhances chemodynamic therapy via glutathione consumption. Moreover, the ONOO - facilitates the upregulation of matrix metalloprotein expression beneficial for extracellular matrix degradation, which in turn enhances the penetration of MNPs and benefits the antitumor CDT/MHT/immuno-related therapy. In vivo experiments have demonstrated an impressive 91.7% inhibition of tumor growth. This hierarchical design offers groundbreaking insights for further advancements in MNP-based tumor therapy. Its implications extend to a broader audience, encompassing those interested in material science, biology, oncology, and beyond., Competing Interests: Declaration of Competing Interest The authors frankly declare that there was no conflict of interest regarding the publication of this article., (Copyright © 2023. Published by Elsevier Ltd.)

Chemodynamic therapy (CDT), as an emerging therapeutic strategy, kills cancer cells by converting intracellular hydrogen peroxide (H 2 O 2 ) into cytotoxic oxidizing hydroxyl radicals (⋅OH). However, the therapeutic efficiency of CDT is compromised due to the insufficient endogenous H 2 O 2 and metal catalysts in tumor cells. The use of multivalent polyphenols with multiple hydroxyl functions provides a facile yet robust means for efficient CDT augmentation. For this purpose, we reported herein the construction of polyphenol-metal nanoparticles (NPs) via a phenol-metal coordination strategy. The uniqueness of this study is the preparation of only one polymer construct with multivalency that can afford various supramolecular interactions for simultaneous "one-pot" loading of different therapeutic species, i.e., doxorubicin (DOX), glucose oxidases (GOD), and Fe 3+ and further co-self-assembly into a stabilized nanomedicine for cascade amplified chemo-chemodynamic therapy. Specifically, the tumor intracellular acidic pH-triggered DOX release could serve for chemotherapy as well as enhance the intracellular H 2 O 2 level. Together with the extra H 2 O 2 and gluconic acid produced by the GOD-triggered glucose consumption, DOX@POAD-Fe@GOD NPs promoted Fe 3+ participation in the Fe-mediated Fenton reaction for cascade amplified chemo-chemodynamic therapy. Notably, this formulation displayed a greater anti-tumor effect with a tumor inhibition ratio 1.6-fold higher than that of free DOX in a BALB/c mice model bearing 4T1 tumors. Overall, the multivalent polyphenol-metal nanoplatform developed herein integrates chemotherapy, starvation therapy, and CDT for synergistic enhanced anticancer efficiency, which shows great potential for clinical translations. STATEMENT OF SIGNIFICANCE: Chemodynamic therapy (CDT) generally suffers from compromised therapeutic efficiency due to insufficient endogenous H 2 O 2 and metal catalysts in tumor cells. To develop a facile yet robust strategy for efficient CDT augmentation, we reported herein construction of a multivalent polyphenol-metal nanoplatform, DOX@POAD-Fe@GOD nanoparticles (NPs) via a phenol-metal coordination strategy. This nanoplatform integrates multiple supramolecular dynamic interactions not only for simultaneously safe encapsulation of doxorubicin (DOX), Fe 3+ , and glucose oxidases (GOD), but also for cascade amplified chemo-chemodynamic therapy. Specifically, the intracellular acidic pH-triggered dissociation of DOX@POAD-Fe@GOD NPs promoted the release of Fe 3+ , DOX, and GOD for significantly increased ROS levels that can accelerate Fenton reactions for cascaded chemotherapy, starvation therapy, and CDT with amplified antitumor efficiency in vivo., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interest., (Copyright © 2023 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Immunotherapy is an emerging treatment modality for tumors after surgery, radiotherapy, and chemotherapy. Despite the potential for eliminating primary tumor cells and depressing cancer metastasis, immunotherapy has huge challenges including low tumor immunogenicity and undesirable immunosuppressive tumor microenvironment (TME). Herein, the two-pronged microenvironmental modulation nanoplatform is developed to overcome these limitations. Specifically, hollow mesoporous MnO 2 (HM) nanoparticles with pH responsive property are prepared and modified with glucose oxidase (GOX) by amide bond, which are further loaded with a potent glutaminase inhibitor CB839 to obtain HM-GOX/CB839. Under the low pH values in TME, HM was disintegrated, thereby releasing Mn 2+ , GOX and CB839. On the one hand, Mn 2+ can convert H 2 O 2 that increased by GOX catalysis in tumors into highly toxic hydroxyl radicals (•OH) and further induce immunogenic cell death (ICD) through the metal-oxidase cascade catalytic reaction, enhancing immunogenicity. On the other hand, GOX and CB839 can block glycolytic and glutamine metabolism pathways, respectively, which effectively reduce the number of immunosuppressive cells and reshape TME, improving anti-tumor immune efficacy. It is demonstrated that HM-GOX/CB839 can effectively activate the body's immunity and inhibit tumor growth and metastasis, providing a potential strategy for comprehensive tumor therapy. STATEMENT OF SIGNIFICANCE: Integrated microenvironmental modulation of metal-oxidase cascade catalysis and metabolic intervention offers a potential avenue for tumor immunotherapy. Under this premise, we constructed a two-pronged microenvironmental modulation nanoplatform (HM-GOX/CB839). On the one hand, the metal oxidase cascade could catalyze the generation of hydroxyl radicals (•OH) and induce immunogenic cell death (ICD), enhancing immunogenicity; on the other hand, metabolic intervention reprogrammed tumor microenvironment to relieve immunosuppression and thereby enhancing anti-tumor immune response. The resulting data demonstrated that HM-GOX/CB839 effectively inhibited tumor growth and metastasis, providing therapeutic potential for cancer immunotherapy., Competing Interests: Declaration of Competing Interest All the authors have read, approved this manuscript and declare no conflict of interest., (Copyright © 2023. Published by Elsevier Ltd.)

The establishment of sensitive and facile colorimetric platform based on the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is of great significance for in vitro diagnosis. Herein, we develop a dual-enzyme cascade amplification strategy based on CRISPR-Cas12a and glucose oxidase (GOx) for instrument-free and sensitive detection of target analytes. HPV-16 DNA as the model nucleic acid target directly initiated CRISPR-Cas12a-based signal transduction, resulting in the enzymatic cleavage of ssDNA linker and the release of GOx from magnetic nanoparticles 1 (MNPs1). Following simple magnetic separation, the supernatant containing GOx was taken out and used to catalyze the substrate, resulting in a visually detectable color change. The detection limit (LOD) of HPV-16 DNA was as low as 1 pM, and the entire process could be completed within 70 min without the need for expensive equipment. Notably, the dual-enzyme cascade amplification strategy was successfully applied to the detection of non-nucleic acid targets, such as ATP, via a simple signal transduction process. The visual LOD for ATP detection reaches 2.5 μM. The approach provides a robust, sensitive and reliable point-of-care biosensing platform for the detection of target analytes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This work reports for the first time a colorimetric based biosensor using nanocellulose (NC) based supports drop-deposited onto a cellulose paper substrate for glucose detection in point-of-care. For this purpose, microcrystalline cellulose (MCC) samples were oxidized with 2,2,6,6-tetramethylpiperidine-N-oxyl radical (TEMPO), sodium hypochlorite, and potassium bromide, to produce carboxylated NC. For the characterization, we used several methods: TEM, FTIR and conductometric titration. In all samples, the primary alcohol groups were selectively oxidized into carboxyl groups, provided the sodium hypochlorite is added dropwise and the reaction is performed at constant pH 10. Carboxyl- NC was further casted on a cellulose substrate and used as support for glucose oxidase (GOx), horseradish peroxidase (HRP) and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) reactions, aiming to yield a coloured detection system for glucose.The sensing system was generated by integrating GOx on the carboxyl- NC /cellulose substrate. Upon reaction with glucose, the enzyme produced hydrogen peroxide, which was converted into a blue-coloured product by reaction with HRP and the chromogenic reagent ABTS. The test-strip was calibrated by incubating it in different concentrations of glucose. The colours obtained were further analysed by a suitable image analysis software. Linear response for glucose ranged from 1.5 to 13.0 mM.Overall, this new test-strip used renewable material for glucose determination, which is an advantage when compared to other systems that require more complex technological approaches. Moreover, it was found that carboxyl- NC improved the colour homogeneity of the test-strip and the intrinsic linear response of concentration range.

In this work, a novel "turn-on" fluorescence sensor for the detection of H 2 O 2 and glucose was developed based on green fluorescent carbon dots (CDs). The CDs was newly prepared by a facile one-pot hydrothermal method with Eosin Y and branched polyethylenimine as precursors. Interestingly, in the presence of H 2 O 2 and HRP, the fluorescence of the CDs enhanced significantly with a red-shift emission due to their "aggregation". Meanwhile, the oxidation of glucose catalyzed by glucose oxidase could generate H 2 O 2 . Thus, a simple sensing system based on the CDs as fluorescent probes was constructed for H 2 O 2 and glucose determination, avoiding the fluorescence quenching and subsequent recovery process in conventional turn-on strategy. The method showed good selectivity and sensitivity for glucose sensing with the detection limit of 0.12 μM. The method was further applied to glucose detection in real samples. The obtained results demonstrated the simplicity, selectivity and practicality of the method. This work expands the carbon nanomaterials with fluorescence emission enhancement properties. It provides a new and direct "turn-on" strategy for H 2 O 2 and glucose detection, which could be a simple and effective tool for screening biological substances involved in H 2 O 2 -generation reaction., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The detection of human motion and sweat composition are important for human health or sports training, so it is necessary to develop flexible sensors for monitoring exercise processes and sweat detection. Mussel secretion of adhesion proteins enables self-healing of byssus and adhesion to surfaces. We prepared Au nanoparticles@polydopamine (AuNPs@PDA) nanomaterials based on mussel-inspired chemistry and compounded them with polyvinyl alcohol (PVA) hydrogels to obtain PVA/AuNPs@PDA self-healing nanocomposite hydrogels. Dopamine (DA) was coated on the surface of AuNPs to obtain AuNPs based composite (AuNPs@PDA) and the AuNPs@PDA was implanted into the PVA hydrogels to obtain nanocomposite hydrogel through facile freeze-thaw cycle. Glucose oxidase (GOD) was added to the hydrogel matrix to achieve specific detection of glucose in sweat. The obtained hydrogels exhibit high deformability (573.7 %), excellent mechanical strength (550.3 KPa) and self-healing properties (85.1 %). The PVA/AuNPs@PDA hydrogel sensors exhibit quick response time (185.0 ms), wide strain sensing range (0-500 %), superior stability and anti-fatigue properties in motion detection. The detection of glucose had wide concentration detection range (1.0 μmol/L-200.0 μmol/L), low detection limits (0.9 μmol/L) and high sensitivity (24.4 μA/mM). This work proposes a reference method in dual detection of human exercise and sweat composition analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In clinical practice, surgery is the preferred treatment for breast cancer; however, the high recurrence rate due to residual tumors after surgery remains a major issue. Hydrogels can reduce the side effects of residual tumors and exert strong anticancer effects, thereby showing potential as therapeutic agents for suppressing tumor recurrence after surgery. Glucose oxidase (GOD)-immobilized gelatin hydrogels (GOD-gelatin hydrogel) were prepared by bioorthogonal click chemistry. Then, the anticancer effect, tumor recurrence inhibition, and biodegradability of the resulting hydrogels were evaluated through cell and animal experiments. GOD-gelatin hydrogel showed cytotoxicity and anticancer effect via H 2 O 2 generation. Unlike free GOD, GOD-gelatin hydrogel remained in the surgical site after implant and continued to suppress tumor recurrence over time. The proposed GOD-gelatin hydrogel system can be easily implanted at the surgical site after tumor surgery, representing a novel treatment to suppress tumor recurrence without any systemic toxicity., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Changkyu lee has patent #10–2023–0036995 pending to changkyu lee., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The self-powered sensor (SPS) is a sensor method that does not require the external power source and has the potential for portable detection of environmental contaminants. In this work, for the first time, a biomolecule-free SPS for detection of ultra-trace triazine endocrine disruptor atrazine (ATZ) with high sensitivity and selectivity is constructed using a glucose oxidase (GOD)-like cobalt metal-organic framework (Co-MOF) nanozyme-modified high-performance anode and a molecularly imprinted cathode. By modulating the size and morphology of the prepared materials, Co-MOF nanozyme with superior GOD-like property (Michaelis constant K m  = 15.8 mM) has been obtained and modified at the anode to catalyze glucose oxidation with high efficiency and provide energy continuously and stably for the SPS. The separation mode of anodic energy supply-cathodic recognition ensures the recognition effect without affecting the catalytic characteristic of Co-MOF and the output signal of the SPS. The designed SPS has a wide linear range of 1 pM-100 nM and a detection limit as low as 0.65 pM, as well as superior selectivity and good stability. The present work provides a promising approach for the design of self-powered sensors which can be extended to detection of a wider range of environmental pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In recent years, two-dimensional nanosheet metal-organic frameworks (2D MOFs) have been widely considered as promising carriers for enzyme immobilization owing to their large surface area, designable and tunable structures, and other properties that enhance enzyme loading and modulate interactions with enzymes. In this study, a series of ultrathin 2D M-TCPP (M = Co, Ni, Zn, Cu) nanosheets were synthesized employing a surfactant-assisted bottom-up approach, and the effect of solvent ratio on the morphology and properties of 2D MOFs was explored. After systematic characterization, Cu-based 2D MOFs with large specific surface areas and excellent water stability was selected as the carrier for the co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP). The effects of adsorption and covalent immobilization strategies on bis-enzyme loading and enzyme activity, as well as their applications in rapid glucose detection, were systematically investigated. The results showed that A-CTGH and C-CTGH owned enzyme loadings of 187.9 and 249.1 mg/g, respectively, and exhibited superior enzymatic activity when exposed to harsh environments compared to free enzymes. In addition, the covalently immobilized biocatalyst based on GOx demonstrated a more sensitive glucose detection performance, including a wide linear range from 5.0 to 16 μM with a detection limit of 0.6 μM., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)