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Entrapment of glucose oxidase (GOx) within metallic gold converts this widely used enzyme into a general saccharide oxidase. The following sugar molecules were oxidized by the entrapped enzyme (in addition to D-glucose): fructose, xylose, L-glucose, glucose-6-phosphate, sucrose, lactose, methylglucoside, and the tri-saccharide raffinose. With the exception of raffinose, none of these sugars have a natural specific oxidase. The origin of this generalization of activity is attributed to the strong protein-gold 3D interactions and to the strong interactions of the co-entrapped CTAB with both the gold, and the protein. It is proposed that these interactions induce conformational changes in the channel leading to the active site, which is located at the interface between the two units of the dimeric GOx protein. The observations are compatible with affecting the specific conformation change of pulling apart and opening this gate-keeper, rendering the active site accessible to a variety of substrates. The entrapment methodology was also found to increase the thermal stability of GOx up to 100 °C and to allow its convenient reuse, two features of practical importance., (© 2022. The Author(s).)

Glucose oxidase is a subset of oxidoreductase enzymes that catalyzes the transfer of electrons from an oxidant to a reductant. Glucose oxidases use oxygen as an external electron acceptor that releases hydrogen peroxide (H 2 O 2 ). Glucose oxidase has many applications in commercial processes, including improving the color and taste, increasing the persistence of food materials, removing the glucose from the dried egg, and eliminating the oxygen from different juices and beverages. Moreover, glucose oxidase, along with catalase, is used in glucose testing kits (especially in biosensors) to detect and measure the presence of glucose in industrial and biological solutions (e.g., blood and urine specimens). Hence, glucose oxidase is a valuable enzyme in the industry and medical diagnostics. Therefore, evaluating the structure and function of glucose oxidase is crucial for modifying as well as improving its catalytic properties. Finding different sources of glucose oxidase is an effective way to find the type of enzyme with the desired catalysis. Besides, the recombinant production of glucose oxidase is the best approach to produce sufficient amounts of glucose oxidase for various uses. Accordingly, the study of various aspects of glucose oxidase in biotechnology and bioprocessing is crucial., (© 2021 International Union of Biochemistry and Molecular Biology, Inc.)

The Fenton reaction is regarded as highly efficient for the degradation of organic contaminants. However, the traditional Fenton reaction is still flawed in a narrow pH working range and low utilization efficiency of the reagents. Based on two striking features, a sustained release of H 2 O 2 in-situ under the catalysis of glucose oxidase (GOX) and the rapid electron donation & transferability from green rust (GR), an adaptable biological Fenton-like system (GGGMFs) was established. The coupling roles of glucose, GOX and GR in the degradation of 3,4-dimethylaniline (3,4-DMA) and the types of reactive species were deduced by electron spin resonance (ESR), etc.. Results demonstrated that the suitable pH range of the system was optimized from acidic to circumneutral, which was favorable for practical application, owing to the heterogeneous formation of GR and the pH self-adjustable capacity of GOX-Glucose. Meanwhile, hydroxyl radical (·OH), superoxide radical (·O 2 - ) and Fe (IV) were identified to be the main oxidizing reactive species. Taking different selectivity of the reactive species to certain pollutant functional groups into consideration, the degradation pathways of 3,4-DMA were proposed. Moreover, it was shown that GR not only acted as the activating substance of the Fenton-like reaction, but also enhanced the activity of GOX, resulting in the promotion of glucose conversion in GGGMFs. This study shed light on the enhancement mechanism consisting of two aspects: (i) the elimination of product inhibition (ii) the formation of a 2Fe(III)-FAD complex with FAD, the active center of GOX, which prompted the electronic transfer in the enzyme catalytic reaction., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Life is fueled by multi-enzymatic tandem processes that display unmatched catalytic efficiencies owing to certain features of the biological reactors such as compartmentalization, nano-confinement, and out-of-equilibrium dynamics. With an attempt to match such natural catalytic systems, herein, we present a chemoenzymatic pH clock mediated transient assembly of a vesicular nanozyme. Distinct confinement of two catalytically discrete units, Histidine groups on the periphery and hemin in the lipid bilayer, results in an efficient hydrolase-peroxidase tandem catalysis in a temporally controlled fashion. The pH clock, constituted by alkaline TRIS (Tris(hydroxymethyl)aminomethane hydrochloride) buffer (promoter) and glucose oxidase (GOx) catalyzed oxidation of glucose, steers the transience in an asymmetric fashion. Alkaline TRIS buffer enhances the pH of the system and triggers the formation of imine linked Supramolecular Peptide Amphiphiles (SPAs) that further assemble into vesicles. On the other hand, oxidation of glucose produces gluconolactone and H 2 O 2 . Gluconolactone hydrolyzes to gluconic acid (deactivator) which dissipates the nanozyme while H 2 O 2 is used in the peroxidase catalysis. Thus, the bi-directional feedback from the fuel not only regulates the existence of the transient state but also controls the activity of the assembly. The transiently assembled nanozyme protected the activity of the catalytic units, displayed substrate specificity and catalytic reproducibility over multiple fueling cycles., 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 © 2022 Elsevier Inc. All rights reserved.)

It is generally accepted that glucose oxidase (GOx) shows unique specificity in β-d-glucose catalysis. However, it has been found that GOx can catalyze diverse monosaccharides. Therefore, the sensing accuracy for glucose biosensors using GOx as probes will be largely compromised by the presence of other monosaccharides. Herein, multifunctional bi-nanospheres (Fe 3 O 4 @Au NCs), which show both peroxidase-like and catalase-like catalytic activities in different working conditions, are successfully constructed and served as desirable platform with huge surface area for the immobilization of large amount of GOx probes. In acidic environment, hydroxyl radicals could be generated via the cascaded catalysis of β-d-glucose by Fe 3 O 4 @Au-GOx, and then employed to initiate the polymerization of boric acid derivative to prepare molecularly imprinted polymers (MIPs) on the surface of GOx using β-d-glucose as template. Then, the molecularly imprinted GOx are immobilized on the surface of highly oriented pyrolytic graphite (HOPG) electrode and an electrochemical biosensor (Fe 3 O 4 @Au-GOx-HOPG) for glucose sensing is successfully obtained. Interestingly, the as-prepared biosensors could selectively detect glucose in the range of 10.0 μM - 5.0 mM with a LOD = 5.0 μM with the help of MIPs, which is comparable or better than other glucose sensors reported recently., (Copyright © 2022. Published by Elsevier B.V.)

In recent years, many efforts have been made to develop rapid, sensitive and user-friendly glucose biosensors for monitoring blood glucose concentration in patients. In this study, the electrochemical glucose biosensors based on graphite rod (GR) electrode electrochemically modified with dendritic gold nanostructures (DGNs) and glucose oxidase (GOx) were developed. Phenazine methosulfate was used as a soluble redox mediator. Three GOx immobilization methods: adsorption on DGNs and cross-linking with glutaraldehyde (GA) vapour (GA-GOx/DGNs/GR), covalent immobilization on DGNs modified with 11-mercaptoundecanoic acid self-assembled monolayer (SAM) (GOx-SAM/DGNs/GR) and covalent immobilization on SAM with additional cross-linking with GA vapour (GA-GOx-SAM/DGNs/GR), were used. It was determined that GA significantly improved the stability of the enzyme layer. The difference of maximal current generated during the enzymatic reaction (Δ I max ) equal to 272.06 ± 8.69 µA was obtained using a biosensor based on GA-GOx/DGNs/GR electrodes. However, the highest Δ I max equal to 384.20 ± 16.06 µA was obtained using GA-GOx-SAM/DGNs/GR electrode. Δ I max for biosensors based on the GA-GOx-SAM/DGNs/GR electrode was 1.41 times higher than for the GA-GOx/DGNs/GR, whereas the linear dynamic range from 0.1 to 10 mM was the same using all three GOx immobilization methods. The limit of detection using GA-GOx-SAM/DGNs/GR and GA-GOx/DGNs/GR electrodes was 0.019 and 0.022 mM, respectively. The ability to detect glucose in the serum by developed biosensors was evaluated.

The adoption of existing continuous glucose monitors (CGMs) is limited by user burden. Herein, a design for a glucose biosensor with the potential for subcutaneous implantation, without the need for a transcutaneous probe or affixed transmitter, is presented. The design is based on the combination of an enzyme-driven phosphorescence lifetime-based glucose-sensing assay and a thermoresponsive membrane anticipated to reduce biofouling. The metalloporphyrin, Pd meso-tetra(sulfophenyl)-tetrabenzoporphyrin ([PdPh 4 (SO 3 Na) 4 TBP] 3 , HULK) as well as glucose oxidase (GOx) are successfully incorporated into the UV-cured double network (DN) membranes by leveraging electrostatic interactions and covalent conjugation, respectively. The oxygen-sensitive metalloporphyrin is incorporated at different levels within the DN membranes. These HULK-containing membranes retain the desired thermosensitivity, as well as glucose diffusivity and primary optical properties of the metalloporphyrin. After subsequently modifying the membranes with GOx, glucose-sensing experiments reveal that membranes prepared with the lowest GOx level exhibit the expected increase in phosphorescent lifetime for glucose concentrations up to 200 mg dL -1 . For membranes prepared with relatively higher GOx, oxygen-limited behavior is considered the source of diminished sensitivity at higher glucose levels. This proof-of-concept study demonstrates the promising potential of a biosensor design integrating a specific optical biosensing chemistry into a thermoresponsive hydrogel membrane., (© 2022 Wiley-VCH GmbH.)

Although "closed-loop" smart insulin delivery systems have been extensively investigated, the majority of them suffer from low insulin loading efficiency and slow glucose response. Here, we constructed a novel nanocomplex (NC), which was prepared by electrostatic interaction between negatively charged insulin prodrug nanoparticles (NPs) and positively charged polycaprolactone-polyethylenimine (PCL-PEI) micelles. The insulin prodrug was linked to acetalated dextran (AD) via borate ester bonds to form IAD NPs, and glucose oxidase (GOx) was encapsulated in PCL-PEI micelles. The NC was negatively charged with a high insulin grafting rate (0.473 mg/mg), and in vitro experiments revealed that IAD was sensitive to hyperglycemia and H 2 O 2 , whereas GOx significantly improved the response to glucose by altering the microenvironment to promote sustained insulin release. Furthermore, compared with free insulin and IAD NPs, subcutaneously injected NCs in diabetic rats had long-term hypoglycemic effects, showing excellent biocompatibility in vitro and in vivo, which had good potential in insulin self-regulation delivery.

Colloidal assemblies of mesoporous suprastructures provide effective catalysis in an advantageous volume-confined environment. However, typical fabrication methods of colloidal suprastructures are carried out under toxic or harmful conditions for unstable biomolecules, such as, biocatalytic enzymes. For this reason, biocatalytic enzymes have rarely been used with suprastructures, even though biocatalytic cascade reactions in confined environments are more efficient than in open conditions. Here, multimodal enzyme- and photocatalyst-carrying superstructures with efficient cascade reactions for colorimetric glucose detection are demonstrated. The suprastructures consisting of various functional nanoparticles, including enzyme-carrying nanoparticles, are fabricated by surface-templated evaporation driven suprastructure synthesis on polydimethylsiloxane-grafted surfaces at ambient conditions. For the fabrication of suprastructures, no additional chemicals and reactions are required, which allows maintaining the enzyme activities. The multimodal enzymes (glucose oxidase and peroxidase)-carrying suprastructures exhibit rapid and highly sensitive glucose detection via two enzyme cascade reactions in confined geometry. Moreover, the combination of enzymatic and photocatalytic cascade reactions of glucose oxidase to titanium dioxide nanoparticles is successfully realized for the same assay. These results show promising abilities of multiple colloidal mixtures carrying suprastructures for effective enzymatic reactions and open a new door for advanced biological reactions and enzyme-related works., (© 2021 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Glucose oxidase (GOx) is an important oxidoreductase enzyme with many important roles in biological processes. It is considered an "ideal enzyme" and is often called an oxidase "Ferrari" because of its fast mechanism of action, high stability and specificity. Glucose oxidase catalyzes the oxidation of β-d-glucose to d-glucono-δ-lactone and hydrogen peroxide in the presence of molecular oxygen. d-glucono-δ-lactone is sequentially hydrolyzed by lactonase to d-gluconic acid, and the resulting hydrogen peroxide is hydrolyzed by catalase to oxygen and water. GOx is presently known to be produced only by fungi and insects. The current main industrial producers of glucose oxidase are Aspergillus and Penicillium. An important property of GOx is its antimicrobial effect against various pathogens and its use in many industrial and medical areas. The aim of this review is to summarize the structure, function, production strains and biophysical and biochemical properties of GOx in light of its various industrial, biotechnological and medical applications.

Tumor vasculature-targeting therapy either using angiogenesis inhibitors or vascular disrupting agents offers an important new avenue for cancer therapy. In this work, a tumor-specific catalytic nanomedicine for enhanced tumor ablation accompanied with tumor vasculature disruption and angiogenesis inhibition was developed through a cascade reaction with enzyme glucose oxidase (GOD) modified on Fe-based metal organic framework (Fe-MOF) coupled with anti-VEGFR2.The GOD enzyme could catalyze the intratumoral glucose decomposition to trigger tumor starvation and yet provide abundant hydrogen peroxide as the substrate for Fenton-like reaction catalyzed by Fe-MOF to produce sufficient highly toxic hydroxyl radicals for enhanced chemodynamic therapy and instantly attacked tumor vascular endothelial cells to destroy the existing vasculature, while the anti-VEGFR2 antibody guided the nanohybrids to target blood vessels and block the VEGF-VEGFR2 connection to prevent angiogenesis. Both in vitro and in vivo results demonstrated the smart nanohybrids could cause the tumor cell apoptosis and vasculature disruption, and exhibited enhanced tumor regression in A549 xenograft tumor-bearing mice model. This study suggested that synergistic targeting tumor growth and its vasculature network would be more promising for curing solid tumors. STATEMENT OF SIGNIFICANCE: Cooperative destruction of tumor cells and tumor vasculature offers a potential avenue for cancer therapy. Under this premise, a tumor-specific catalytic nanomedicine for enhanced tumor ablation accompanied with tumor vasculature disruption and new angiogenesis inhibition was developed through a cascade reaction with glucose oxidase modified on the surface of iron-based metal organic framework coupled with VEGFR2 antibody. The resulting data demonstrated that a therapeutic regimen targeting tumor growth as well as its vasculature with both existing vasculature disruption and neovasculature inhibition would be more potential for complete eradication of tumors., 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 © 2022. Published by Elsevier Ltd.)

The integration of multimodal chemo-/bio-catalysis for efficient cascade reactions has long provided broad prospects in the field of biotechnology for ages. In this work, we describe the synthesis of a biomimetic multienzyme hybrid with hierarchically porous structure and outstanding catalytic activity via in situ encapsulation of natural enzymes in an iron-cobalt bimetallic metal-organic framework (Fe/Co-MOF, FCM). The combination of a single enzyme (glucose oxidase) or dual enzyme (β-galactosidase and glucose oxidase) with FCM resulted in remarkable synergistic biocatalysis ability; in contrast to simple biocatalyst mixtures in solution, the prepared multienzyme hybrid resulted in 3.2-fold and 2.1-fold improvements in activity for tandem reactions, respectively. The reinforced cascade bioactivity of the multienzyme hybrid benefitted from the synergistic effect between iron/cobalt in the FCM nanozyme, the opened substrate channel between enzymes/nanozymes, and the beneficial effect provided by the hierarchical MOF pores. The enlarged pores not only provided adequate space for immobilized proteins to diffuse and reorientate in FCM with low surface energy, but also reduced the intrinsic mass transfer obstacle to increase the diffusional efficiency of reactants/intermediates. In addition, on account of the shielding effect provided by FCM, the multienzyme hybrid exhibited enhanced tolerance towards severe circumstances and excellent reusability and has been successfully applied in small molecule detection, such as glucose and lactose. The current study highlights the superiority of synergistic bioreactors integrated with the MOF nanozyme and natural enzymes, suggesting great potential for applications in sustainable biomimetic catalysis.

Pyrite (PR), as a representative sulfide mineral, possesses the advantages of abundancy, thermodynamic stability, non-toxicity and semi-conductivity. In this study, an amperometric glucose biosensor (GOD/CS/PR/GCE) based on layer-by-layer of glucose oxidase (GOD), chitosan (CS) and pyrite (PR) on a glassy carbon electrode (GCE) was fabricated through electrostatic force. In this research, PR suspension prepared in phosphate buffer (pH 5.5) was first immobilized on the GCE surface, which exhibits a negative charge. Then, positively charged CS was adsorbed on the PR/GCE by electrostatic force. Finally, negatively charged GOD was further modified on the CS/PR/GCE surface through electrostatic force again. The surface morphology and adsorbance mechanism were supported by field emission scanning electron microscopy, quartz crystal microbalance with dissipation and atomic force microscope. The step-by-step procedure gives both strong adhesion ability and good bioelectrocatalytic activity of GOD on the CS- and PR-modified electrode surface. The linear range of this GOD/CS/PR/GCE biosensor was achieved from 0.5 to 60 mM with the linear regression equation of y = 0.897x - 0.3016 (R 2  = 0.9996) and a limit of detection value of 50 µM. This approach of using pyrite and chitosan as physically modified GOD to serve as electrostatic glues could be useful for designing better enzyme-based biosensors for a wide variety of practical applications., (© 2022. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

Glyconanoparticles (GNPs) made by self-assembly of carbohydrate-based polystyrene-block-β-cyclodextrin copolymer are used as a building block for the design of nanostructured biomaterials of electrode. The firm immobilization of GNPs is carried out on electrochemically generated polymer, poly(pyrrole-adamantane), and copolymer, poly(pyrrole-adamantane)/poly(pyrrole-lactobionamide) via host-guest interactions between adamantane and β-cyclodextrin. The ability of GNPs for the specific anchoring of biological macromolecules is investigated using glucose oxidase enzyme modified by adamantane groups as a protein model (GOx-Ad). The immobilization of GOx-Ad is carried out by incubation of an aqueous enzyme solution on a coating of GNPs adsorbed on a platinum electrode. The presence of immobilized GOx-Ad is evaluated in aqueous glucose solution by potentiostating the underlying platinum electrode at 0.7 V/SCE for the electro-oxidation of H 2 O 2 generated by the enzyme. The analytical performance of the bioelectrodes for the detection of glucose is compared to control electrodes prepared without GNPs or without electropolymerized films. The better permeability of copolymer compared to polymer and the possibility to elaborate two alternating layers of GNPs and GOx-Ad are clearly observed. The best amperometric response is recorded with a multilayered bioelectrode displaying a wide linear range linear range of the calibration curve: 68 µmol L -1 to 0.1 mol L -1 ., (© 2022 Wiley-VCH GmbH.)

The development of modern technologies has acclimatized biosensors to complicated applicable scenarios with integrated properties as a whole instead of the pursuit of a single-point breakthrough. Here, we targeted a few concerns in the development of enzyme-based biosensors, including stability, analyte enrichment, and signal transduction, and developed a general biosensing model utilizing enzymes, aggregation-induced emission (AIE) luminogens, and stimuli-responsive framework materials as the units. We propose such proof-of-concept of glucose biosensors by coencapsulating glucose oxidase and AIE-type gold nanoclusters into acid-sensitive zeolite imidazolate framework (ZIF)-8 nanocrystals. The acid-activated degradation of ZIF-8 bridges the molecular signals produced by the enzyme-catalytic reaction of glucose and the photon signals generated by ZIF-8-induced AIE effects of gold nanoclusters, resulting in the "turn-off" model nanoprobes for glucose detection with high selectivity. After embedding the nanoprobes into hollow-out tapes, the formed paper biosensors can conveniently detect glucose with the help of a smartphone.

The unrestrained use of antibiotics accelerates the development of drug-resistant bacteria and leads to an increasing threat to human health. Therefore, there is an urgent need to explore novel and effective strategies for the treatment of bacterial infections. Herein, zeolite imidazole framework-8 (ZIF-8) material was utilized to construct biomineralized nanomaterial (GOx&HRP@ZIF-8/ASO) by encapsulating biological cascade enzymes and combining with antisense oligonucleotides (ASOs), which achieved effective and synergistic antidrug-resistant bacteria therapy. Various in vitro assays confirmed that GOx&HRP@ZIF-8/ASO exhibited excellent antibacterial properties against Escherichia coli , Staphylococcus aureus , methicillin-resistant S. aureus (MRSA) during catalysis of glucose (Glu), especially the minimum inhibitory concentration (MIC) against MRSA was only 16 μg/mL. Compared with simple ZIF-8 (32.85%) and ftsZ ASO (58.65%), GOx&HRP@ZIF-8/ASO+Glu exhibited superb biofilm destruction ability, and the bacteria removal efficiency of the MRSA biofilm could be as high as 88.2%, indicating that the reactive oxygen species (ROS) produced by the cascade enzyme reaction imparted the main synergistic antibacterial capability, and simultaneously, ftsZ ASO significantly enhanced the antibacterial effect by inhibiting the expression of the ftsZ gene. In vivo anti-infection treatment experiments revealed that GOx&HRP@ZIF-8/ASO exhibited the best wound repairing performance and excellent biocompatibility in the presence of Glu. These findings suggested that GOx&HRP@ZIF-8/ASO has favorably realized high-efficiency treatment of MRSA infection and filled the gap in the antibacterial application of biological enzymes.

In eukaryotic cells, enzymes are compartmentalized into specific organelles so that different reactions and processes can be performed efficiently and with a high degree of control. In this work, we show that these features can be artificially emulated in robust synthetic organelles constructed using an enzyme co-compartmentalization strategy. We describe an in situ encapsulation approach that allows enzymes to be loaded into silica nanoreactors in well-defined compositions. The nanoreactors can be combined into integrated systems to produce a desired reaction outcome. We used the selective enzyme co-compartmentalization and nanoreactor integration to regulate competitive cascade reactions and to modulate the kinetics of sequential reactions involving multiple nanoreactors. Furthermore, we show that the nanoreactors can be efficiently loaded into giant polymer vesicles, resulting in multi-compartmentalized microreactors., (© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Redox enzymes are capable of harvesting electrical energy from biofuels in high catalytic activity and under mild condition. However, it is difficult to achieve efficient electron transfer and deep oxidation of biofuels simultaneously in a single-enzyme catalytic system. Herein, we report a hybrid catalyst cascade consisting of an organic oxidation catalyst, 2,2,6,6-tetramethyl-1-piperidine N-oxyl (TEMPO), and an enzyme, glucose oxidase (GOx), for electrochemical oxidation of glucose. It is found that TEMPO is capable of mediating electron transfer between the redox center of GOx and the electrode surface. While glucose can be oxidized into glucuronic acid under neutral conditions. Thus, combining GOx and TEMPO, we are able to achieve 4e - electrooxidation of glucose using the hybrid enzymatic and organic cascade (HEOC) system. When coupled with an air-breathing Pt cathode, the resulting glucose/air biofuel cell using the proposed HEOC anode exhibits a maximum power density of 38.1 μW cm -2 with a short-circuit current of 651.4 μA cm -2 , which can be attributed to the enhanced energetic efficiency, enabling TEMPO a promising catalyst for glucose oxidation in bioelectronics 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 © 2021. Published by Elsevier B.V.)

Wearable electronic medical devices measuring continuous biological signals for early disease diagnosis should be small and lightweight for consecutive usability. As a result, there has been an increasing need for new energy supply systems that provide continuous power without any interruption to the operation of the medical devices associated with the use of conventional batteries. In this work, we developed a patch-type self-charging supercapacitor that can measure biological signals with a continuous energy supply without batteries. The glucose oxidase coated on the surface of the microneedle-type glucose sensor encounters glucose in the interstitial fluids of the human body. Electrons created by glucose oxidation operate the self-powered system in which charging begins with the generation of potential differences in supercapacitor electrodes. In an 11 mM glucose solution, the self-powered solid-state supercapacitors (SPSCs) showed a power density of 0.62 mW/cm 2 , which resulted in self-charging of the supercapacitor. The power density produced by each SPSC with a drop of 11 mM glucose solution was higher than that produced by glucose-based biofuel cells. Consequently, the all-in-one self-powered glucose sensor, with the aid of an Arduino Uno board and appropriate programming, effectively distinguished normal, prediabetic, and diabetic levels from 0.5 mL of solutions absorbed in a laboratory skin model.

Background: Cancer is the most serious world's health problems on the global level and various strategies have been developed for cancer therapy. Pillar[5]arene-based supramolecular therapeutic nano-platform (SP/GOx NPs) was constructed successfully via orthogonal dynamic covalent bonds and intermolecular H-bonds with the assistance of glucose oxidase (GOx) and exhibited efficient targeted/synergistic chemo-chemodynamic cancer therapy., Methods: The morphology of SP/GOx NPs was characterized by DLS, TEM, SEM and EDS mapping. The cancer therapy efficinecy was investigated both in vivo and in vitro., Results: SP/GOx NPs can load drug molecules (Dox) and modify target molecule (FA-Py) on its surface conveniently. When the resultant FA-Py/SP/GOx/Dox NPs enters blood circulation, FA-Py will target it to cancer cells efficiently, where GOx can catalyst the overexpressed glucose to generate H 2 O 2 . Subsequently, the generated H 2 O 2 in cancer cells catalyzed by ferrocene unit to form •OH, which can kill cancer cells. Furthermore, the loaded Dox molecules released under acid microenvironment, which can further achieve chemo-therapy., Conclusion: All the experiments showed that the excellent antitumor performance of FA-Py/SP/GOx/Dox NPs, which provided an new method for pillar[5]arene-based supramolecular polymer for biomedical applications., (© 2022. The Author(s).)

The commonly required properties of diffusive electron mediators for point-of-care testing are rapid dissolubility, high stability, and moderate formal potential in aqueous solutions. Inspired by nature, various quinone-containing electron mediators have been developed; however, satisfying all these requirements remains a challenge. Herein, a strategic design toward quinones incorporating sulfonated thioether and nitrogen-containing heteroarene moieties as solubilizing, stabilizing, and formal potential-modulating groups is reported. A systematic investigation reveals that di(thioether sulfonate)-substituted quinoline-1,4-dione (QLS) and quinoxaline-1,4-dione (QXS) display water solubilities of ≈1 m and are rapidly dissoluble. By finely balancing the electron-donating effect of the thioethers and the electron-withdrawing effect of the nitrogen atom, formal potentials suitable for electrochemical biosensors are achieved with QLS and QXS (-0.15 and -0.09 V vs Ag/AgCl, respectively, at pH 7.4). QLS is stable for >1 d in PBS (pH 7.4) and for 1 h in tris buffer (pH 9.0), which is sufficient for point-of-care testing. Furthermore, QLS, with its high electron mediation ability, is successfully used in biosensors for sensitive detection of glucose and parathyroid hormone, demonstrating detection limits of ≈0.3 × 10 -3 m and ≈2 pg mL -1 , respectively. This strategy produces organic electron mediators exhibiting rapid dissolution and high stability, and will find broad application beyond quinone-based biosensors., (© 2021 Wiley-VCH GmbH.)

In recent years, three key techniques including random co-immobilization, positional co-immobilization, and compartmentalization for multi-enzyme immobilization were extensively considered. Herein, we investigate random co-immobilization and positional co-immobilization techniques for multi-enzyme systems in detail. We describe randomly co-immobilized glucose oxidase (GOx) and horseradish peroxidase (HRP) on reduced graphene oxide (rGO) as the most used methods. Materials and methods are presented in terms of preparation of GO and rGO as well as enzyme immobilization procedure. Moreover, the principles of positional co-immobilization have been reviewed, and the relevant methods based on microfluidic systems and DNA structure considering HRP and GOx enzymes have been individually studied. It is believed that the benefits of using the methods associated with random and specifically positional immobilized multi-enzyme systems include not only enhanced cascade enzymatic activity via manipulated surface such as microfluidic systems (including porous materials) and DNA structure but also improved enzyme stability and ease of recovery for recycle., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Cascade reactions catalyzed by multi-enzymatic systems have attracted enormous scientific interest over the last decade. They are an emerging technology that significantly expands the applicability of biocatalysts in several biotechnological processes, such as the synthesis of high value-added products. Immobilization of enzymes on a solid carrier is a commonly used strategy to improve the stability and reuse of multiple enzyme systems. Magnetic nanoparticles have been applied as promising nanocarriers for either the immobilization of one enzyme or the co-immobilization of multiple enzymes. In this chapter, we describe the preparation of magnetic iron oxide nanoparticles γ-Fe 2 O 3 modified with 3-(aminopropyl)-triethoxysilane (APTES), for the simultaneous covalent co-immobilization of oxidoreductases and hydrolytic enzymes, such as cellulase, β-glucosidase (bgl), glucose oxidase (GOx), and horseradish peroxidase (HRP). Several spectroscopic techniques that are used to characterize the structure and the catalytic performance of such systems are also described., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

This chapter reports a single-step preparation of nanoarmored bi-enzyme systems assembled on 1-D and 2-D nanomaterials, with glucose oxidase and peroxidase enzymes as model systems for cascade bio-catalysis. This is a simple and facile method to both exfoliate the bulk 1D (carbon nanotubes, CNT) and 2D nanomaterials (α-Zirconium phosphate, α-ZrP) and bind the enzymes in a single step. Exfoliation of the bulk material enhances the accessible surface area of the materials for the enzyme binding, and it also boosts the diffusion of reagents from the bulk phase to the active sites of the bio-catalysts. For example, a mixture of horseradish peroxidase, glucose oxidase, and bovine serum albumin (BSA) were adsorbed on the surfaces of the α-ZrP nanoplates or carbon nanotubes (CNT) as the bulk materials are exfoliated simultaneously, in a one-step process. The resulting bio-catalysts were thoroughly characterized by powder X-ray diffraction, electron microscopy, biochemical and biophysical methods, while enzyme activity studies proved successful binding of enzymes with retention of activities or even enhancements in their specific activities. For example, GOx/HRP/BSA/CNT displayed 6 times the activity of a mixture of GOx/HRP/BSA, under otherwise identical conditions. Similarly, GOx/HRP/BSA/ZrP had 3.5 times the activity of the corresponding mixture of GOx/HRP/BSA, in the absence of the nanoplates. These robust nano-dispersions worked extraordinarily well as active bio-catalysts. These two kinds of fabricated biocatalyst dispersions are also highly stable., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Understanding the levels of glucose (G) and lactate (L) in blood can help us regulate various chronic health conditions such as obesity. In this paper, we introduced an enzyme-based electrochemical biosensor adopting glucose oxidase and lactate oxidase on two working screen-printed carbon electrodes (SPCEs) to sequentially determine glucose and lactate concentrations in a single drop (~30 µL) of whole blood. We developed a diet-induced obesity (DIO) mouse model for 28 weeks and monitored the changes in blood glucose and lactate levels. A linear calibration curve for glucose and lactate concentrations in ranges from 0.5 to 35 mM and 0.5 to 25 mM was obtained with R-values of 0.99 and 0.97, respectively. A drastic increase in blood glucose and a small but significant increase in blood lactate were seen only in prolonged obese cases. The ratio of lactate concentration to glucose concentration (L/G) was calculated as the mouse's gained weight. The results demonstrated that an L/G value of 0.59 could be used as a criterion to differentiate between normal and obesity conditions. With L/G and weight gain, we constructed a diagnostic plot that could categorize normal and obese health conditions into four different zones. The proposed dual electrode biosensor for glucose and lactate in mouse whole blood showed good stability, selectivity, sensitivity, and efficiency. Thus, we believe that this dual electrode biosensor and the diagnostic plot could be used as a sensitive analytical tool for diagnosing glucose and lactate biomarkers in clinics and for monitoring obesity.

In this study, designing of a stable electrospun blended chitosan (CS)-poly(vinyl alcohol) (PVA) nanofibers for colorimetric glucose biosensing in an aqueous medium was investigated. CS and PVA solutions were blended to acquire an optimum content (CS/PVA:1/4) and electrospunned to obtain uniform and bead-free CS/PVA nanofiber structures following the optimization of the electrospinning parameters (33 kV, 20 cm, and 1.2 ml.h -1 ). Crosslinking process applied subsequently provided mechanically and chemically stable nanofibers with an average diameter of 378 nm. The morphological homogeneity, high fluid absorption ability (>%50), thermal (<230 °C) and morphological stability, surface hydrophilicity and degrability properties of cross-linked CS/PVA nanofiber demonstrated their great potential to be developed as an eye-readable strip for biosensing applications. The glucose oxidase (GOx) and horseradish peroxidase (HRP) was immobilized by physical adsorption on the cross-linked CS/PVA nanofiber. The glucose assay analysis by ultraviolet-visible (UV-Vis) spectrophotometry using the same enzymatic system of the proposed glucose strips in form of absorbance versus concentration plot was found to be linear over a glucose concentration range of 2.7 to 13.8 mM. The prepared naked eye colorimetric glucose detection strips, with lower detection limit of 2.7 mM, demonstrated dramatic color change from white (0 mM) to brownish-orange (13.8 mM). The developed cross-linked CS/PVA nanofiber strips, prepared by electrospinnig procedure, could be easily adapted to a color map, as an alternative material for glucose sensing. Design of a practical, low-cost, and environmental-friendly bio-based CS/PVA testing strips for eye readable detection were presented and suggested as an applicable medium for a wide range of glucose concentrations., (Copyright © 2021 Elsevier B.V. All rights reserved.)

As a class of functional proteins, enzymes possess inherent insignificant features, for instance, mediocre stability and membrane impermeability and reduced enzymatic activity after modification, which partly limit their biomedical applications. Thus, it is indispensable to exploit robust nanoreactors with high enzymatic activity and good stability and cell permeability. Here, the chiral carbon dots (CDs)-glucose oxidase (GOx) nanoreactors named LGOx and DGOx were constructed by the coassembly of GOx with L/D-CDs, respectively. L/DGOx can significantly enhance the activity of GOx and improve the efficient delivery of GOx to cancer cells. Moreover, these nanoreactors can generate hydrogen peroxide to efficaciously kill cancer cells and restrain tumor growth, and DGOx exhibits higher enzymatic activity than LGOx. According to our understanding, this is the first report about utilizing chiral CDs as vectors to construct effective CDs-enzyme nanohybrids for cancer therapy, which is envisioned to be a versatile strategy for multitudinous biomedical applications.

Anticancer therapies with profound efficacy but negligible toxicity are a fundamental pursuit that has been made humanly possible through either targeting or tumor-selective therapeutic (TST) approaches. Herein, we developed a targeting-enhanced tumor-selective cancer therapy aimed at integrating the two approaches by preparing cerium oxide (CeO 2 ) nanocubes with glucose oxidase (GOx) modified on the cube surface and cancer cell membrane (CCM) camouflaged outside. The immune escape and homotypic binding of camouflaged CCM enable targeted delivery of the resultant CeO 2 -GOx@CCM nanoparticles mostly into cancer tissue, while its acidic environment (pH < 6.6) activated a cascade reaction, in which the glucose was first catalyzed by GOx into H 2 O 2 and then by CeO 2 into highly cytotoxic ˙OH killing cancer cells. In the case of off-targeting, when very few CeO 2 -GOx@CCM nanoparticles were accidentally delivered into normal tissue, its neutral pH environment (pH = 7.4) triggered a protective reaction, in which the H 2 O 2 generated was catalyzed by CeO 2 into non-toxic H 2 O and O 2 . Both in vitro and in vivo results demonstrated that this targeting-enhanced TST achieved the most remarkable antitumor performance with negligible toxic side effects.

A chemiluminescence (CL) sensor was constructed for the one-step determination of glucose. Glucose oxidase (GOx) was successfully encapsulated into Zn-doped zeolitic imidazolate framework-67 (Zn-ZIF-67) via a simple one-pot strategy. The as-prepared GOx@Zn-ZIF-67 nanocomposite can trigger cascade reactions of glucose oxidation to generate H 2 O 2 and H 2 O 2 -mediated luminol reaction to give an intense CL emission. The sensor responds linearly to glucose in the 20.0-400.0 μmol·L -1 range with a limit of detection (LOD) of 4.7 μmol·L -1 . Eleven replicated measurements of 200.0 μmol·L -1 glucose solution gives a relative standard deviation (RSD) of 1.7%. The sensor exhibits good selectivity and stability and was successfully applied to the determination of glucose in real human serum samples. Schematic representation of one-step determination of serum glucose with GOx@Zn-ZIF-67 nanocomposite triggering cascade reactions between luminol and glucose., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Therapeutic nanosystems triggered by a specific tumor microenvironment (TME) offer excellent safety and selectivity in the treatment of cancer by in situ conversion of a less toxic substance into effective anticarcinogens. However, the inherent antioxidant systems, hypoxic environment, and insufficient hydrogen peroxide (H 2 O 2 ) in tumor cells severely limit their efficacy. Herein, a new strategy has been developed by loading the chemotherapy prodrug disulfiram (DSF) and coating glucose oxidase (GOD) on the surface of Cu/ZIF-8 nanospheres and finally encapsulating manganese dioxide (MnO 2 ) nanoshells to achieve efficient DSF-based cancer chemotherapy and dual-enhanced chemodynamic therapy (CDT). In an acidic TME, the nanocatalyst can biodegrade rapidly and accelerate the release of internal active substances. The outer layer of MnO 2 depletes glutathione (GSH) to destroy the reactive oxygen defensive mechanisms and achieves continuous oxygen generation, thus enhancing the catalytic efficiency of GOD to burst H 2 O 2 . Benefiting from the chelation reaction between the released Cu 2+ and DSF, a large amount of cytotoxic CuET products is generated, and the Cu + are concurrently released, thereby achieving efficient chemotherapy and satisfactory CDT efficacy. Furthermore, the release of Mn 2+ can initiate magnetic resonance imaging signals for the tracking of the nanocatalyst.

Polyaniline (PANI) was functionalized by thiol-ene click chemistry to obtain carboxylic acid-tethered polyaniline (PCOOH). The versatility of PCOOH as an immobilization matrix was demonstrated by constructing four different biosensors for detection of metabolites and cancer biomarker. Immobilization efficiency of PCOOH was investigated by surface plasmon resonance and fluorescence microscopic analysis which revealed dense immobilization of biomolecules on PCOOH as compared to conventional PANI. A sandwich electrochemical biosensor was constructed using PCOOH for detection of liver cancer biomarker, α-fetoprotein (AFP). The sensor displayed sensitivity of 15.24 µA (ng mL -1 ) -1  cm -2 , with good specificity, reproducibility (RSD 3.4%), wide linear range (0.25-40 ng mL -1 ) at - 0.1 V (vs. Ag/AgCl), and a low detection limit of 2 pg mL -1 . The sensor was validated by estimating AFP in human blood serum samples where the AFP concentrations obtained are consistent with the values estimated using ELISA. Furthermore, utilization of PCOOH for construction of enzymatic biosensor was demonstrated by covalent immobilization of glucose oxidase, uricase, and horseradish peroxidase (HRP) for detection of glucose, uric acid, and H 2 O 2 , respectively. The biosensors displayed reasonable sensitivity (50, 148, 127 µA mM -1  cm -2 ), and linear ranges (0.1-5, 0.1-6, 0.1-7 mM) with a detection limit of 10, 1, and 8 µM for glucose, uric acid, and H 2 O 2 , respectively. The present study demonstrates the capability of PCOOH to support and enable oxidation of H 2 O 2 generated by oxidase enzymes as well as HRP enzyme catalyzed reduction of H 2 O 2 . Thus, PCOOH offers a great promise as an immobilization matrix for development of high-performance biosensors to quantify a variety of other disease biomarkers. Carboxylic acid-tethered polyaniline synthesized by thiol-ene click chemistry was used as matrix to construct four different electrochemical biosensors for detection of cancer biomarker α-fetoprotein, glucose, uric acid, and H 2 O 2 ., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

This work focuses on the synthesis of novel modified core-shell CdTe/CdS quantum dots (QDs) and develops as a fluorescence sensor for glucose determination. The (E)-2,2'-(4,4'-dioxo-2,2'-dithioxo-2H,2'H-[5,5'-bithiazolylidene]-3,3'(4H,4'H)-diyl)bis(3- mercaptopropanoic acid) (DTM) as a new derivative of thiazolidine was synthesized and characterized and used to surface-modification of CdTe/CdS QDs. DTM-capped CdTe/CdS QDs used to immobilization of glucose oxidase (GOD). The intensity fluorescence emission of the CdSe/CdS-DTM/GOD is highly sensitive to the concentration of H 2 O 2 as a byproduct of the catalytic oxidation of glucose. The experimental results showed that the quenched fluorescence was proportional to the glucose concentration within the range of 10 nM-0.32 μM under optimized experimental conditions. The limit of detection of this system was found to be 4.3 nM. Compared with most of the existing methods, this newly developed system possesses many advantages, including simplicity, low cost, and good sensitivity., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Catalytic cascades have drawn much attention by avoiding the isolation of intermediates and due to high atom economy. Yet, developing an efficient, one-pot biocatalytic cascade remains challenging. Combined with the selectivity of biological enzymes and tunable activity of nanozymes, we herein demonstrate an effective bio-nanozyme cascade formed by glucose oxidase (GOx) and in situ-generated nanoceria. The prepared H 2 O 2 -nanoceria complex shows strong oxidative activity for common chromogenic substrates under physiological conditions, which are the optimal reaction conditions for most biological enzymes. Interestingly, GOx not only provides H 2 O 2 for the second step reaction but also simultaneously leads to 7.4-fold enhancement of activity. We characterized the process of in situ generation of nanoceria at pH 7.0 and how proteins boost the activity by enhancing product desorption. In addition, the proposed one-pot bio-nanozyme cascade shows high stability and analytical performance for serum glucose with a detection limit of 5 μM.

Herein, we report the first example of using mesoporous hydrogen-bonded organic frameworks (MHOFs) as the protecting scaffold to organize a biocatalytic cascade. The confined microenvironment of MHOFs has robust and large transport channels, allowing the efficient transport of a wide range of biocatalytic substrates. This new MHOF-confined cascade system shows superior activity, extended scope of catalytic substrates, and ultrahigh stability that enables the operation of complex chemical transformations in a porous carrier. In addition, the advantages of MHOF-confined cascades system for point-of-care biosensing are also demonstrated. This study highlights the advantages of HOFs as scaffold for multiple enzyme assemblies, which has huge potential for mimicking complex cellular transformation networks in a controllable manner., (© 2021 Wiley-VCH GmbH.)

Glucose oxidase-mediated starvation therapy that effectively cuts off energy supply holds great promise in cancer treatment. However, high glutathione (GSH) contents and anoxic conditions severely reduce therapy efficiency and cannot fully kill cancer cells. Herein, to resolve the above problem, this study constructed a biomimetic nanosystem based on nanreproo-MnO 2 with porous craspedia globose-like structure and high specific surface area, and it was further modified with dopamine and folic acid to guarantee good biocompatibility and selectivity toward cancer cells. This nanosystem responsively degraded and reacted with GSH and acid to regenerate O 2 , which significantly increased intracellular O 2 levels, accelerated glucose consumption, and improved starvation therapy efficiency. Moreover, anticancer drug of camptothecin was further loaded, and notably enhanced cancer growth inhibition was obtained at very low drug concentrations. Most importantly, this novel therapy could unprecedentedly inhibit cancer cell migration to a very low ratio of 19%, and detailed cell apoptosis analyses revealed late stage apoptosis contributed most to the good therapeutic effect. This work reported a new train of thought to improve starvation therapy in biomedicine, and provided a new strategy to design targeted nanocarrier to delivery mixed drugs to overcome the restriction of starvation therapy and develop new therapy patterns., (© 2021 IOP Publishing Ltd.)

The development of versatile nanotheranostic agents has received increasing interest in cancer treatment. Herein, in this study, we rationally designed and prepared a novel flowerlike multifunctional cascade nanoreactor, BSA-GOx@MnO 2 @FePt (BGMFP), by integrating glucose oxidase (GOx), manganese dioxide (MnO 2 ) and FePt for synergetic cancer treatment with satisfying therapeutic efficiency. In an acidic environment, intratumoral H 2 O 2 could be decomposed to O 2 to accelerate the consumption of glucose catalyzed by GOx to induce cancer starvation. Moreover, the accumulation of gluconic acid and H 2 O 2 generated along with the consumption of glucose would in turn promote the catalytic efficiency of MnO 2 and boost O 2 evolution, which could enhance the efficiency of starvation therapy. Moreover, FePt as an excellent Fenton agent could simultaneously convert H 2 O 2 to the toxic hydroxyl radical (˙OH), subsequently resulting in amplified intracellular oxidative stress and cell apoptosis. Therefore, BGMFP could catalyze a cascade of intracellular biochemical reactions and optimize the unique properties of MnO 2 , GOx and FePt via mutual promotion of each other to realize O 2 supply, chemodynamic therapy (CDT) and starvation therapy. The anticancer results in vitro and in vivo demonstrated that BGMFP possessed remarkable tumor inhibition capacity through enhancing the starvation therapy and CDT. It is appreciated that BGMFP could be a promising platform for synergetic cancer treatment.

Three-dimensional (3D) printing has showed great potential for the construction of electrochemical sensor devices. However, reported 3D-printed biosensors are usually constructed by physical adsorption and needed immobilizing reagents on the surface of functional materials. To construct the 3D-printed biosensors, the simple modification of the 3D-printed device by non-expert is mandatory to take advantage of the remote, distributed 3D printing manufacturing. Here, a 3D-printed electrode was prepared by fused deposition modeling (FDM) 3D printing technique and activated by chemical and electrochemical methods. A glucose oxidase-based 3D-printed nanocarbon electrode was prepared by covalent linkage method to an enzyme on the surface of the 3D-printed electrode to enable biosensing. X-ray photoelectron spectroscopy and scanning electron microscopy were used to characterize the glucose oxidase-based biosensor. Direct electrochemistry glucose oxidase-based biosensor with higher stability was then chosen to detect the two biomarkers, hydrogen peroxide and glucose by chronoamperometry. The prepared glucose oxidase-based biosensor was further used for the detection of glucose in samples of apple cider. The covalently linked glucose oxidase 3D-printed nanocarbon electrode as a biosensor showed excellent stability. This work can open new doors for the covalent modification of 3D-printed electrodes in other electrochemistry fields such as biosensors, energy, and biocatalysis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Diabetes is a type of disease that threatens human health, which can be diagnosed based on the level of glucose in the blood. Recently, various MOF-based materials have been developed as efficient electrochemical glucose sensors because of their tunable pore channels, large specific surface area well dispersed metallic active sites, etc. In this review, the significance of glucose detection and the advantages of MOF-based materials for this application are primarily discussed. Then, the application of MOF-based materials can be categorized into two types of glucose sensors: enzymatic biosensors and non-enzymatic sensors. Finally, insights into the current research challenges and future breakthrough possibilities regarding electrochemical glucose sensors are considered.

A simple and efficient method was used to synthesize norepinephrine-induced AuPd aerogels (AuPd-NE) with dual enzyme properties, i.e. glucose oxidase-like property, and peroxidase-like property. Thus, AuPd-NE aerogels can be considered as a tandem nanozyme with tandem enzyme-like activity. In the presence of AuPd-NE aerogels, glucose can be decomposed into gluconic acid and H 2 O 2 . Then, H 2 O 2 will continue to decompose into ·OH and H 2 O. The generated ·OH will oxidize colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue products of ox-TMB. Accordingly, an enzyme-free method based on AuPd-NE aerogels was proposed for sensitive colorimetric detection of glucose. The linear range of the developed method was 30 to 250 μM, and the limit of detection was 10 μM. The method presents reliable applicability for blood glucose detection in human serum samples. This study will deepen the understanding of tandem nanozymes and then rationally design tandem nanozymes for many fascinating biomedical applications. A simple, sensitive and reliable one-pot enzyme-free colourimetric assay for glucose was developed., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The coral-like gold micro/nanostructures were formed onto carbon cloth followed by a Prussian blue (PB) electrochemical deposition to construct a highly sensitive H 2 O 2 biosensor. The SEM image of PB/Au/CC showed the coral-like gold morphology, and EDS and XPS tests also further confirmed the successful loading of Au and PB. The electrochemical tests of PB/Au/CC displayed the electrode possessed excellent performance in sensing H 2 O 2 , which was quantified in the linear range from 0.002 to 13.97 mM at an applied potential of -0.05 V, with a sensitivity of 454.97 μA mM -1 cm -2 and a detection limit of 0.5 μM (S/N = 3). And then a convenient sensing platform was established via the cross-linking enzyme aggregates method, using PB as the mediator to realize the construction of glucose BIOSENSOR GOx EA @PB/Au/CC. The biosensor responded to glucose in the sensitivity of 70.76 μA mM -1 cm -2 within the linear range from 0.05 to 3.15 mM with a detection limit of 10 μM. The sensitivity was much higher than the electrode constructed by the cross-linking enzyme method (GOx@PB/Au/CC), and it was also highly selective, reproducible, and stable. Besides, the proposed biosensor was successfully applied to the glucose determination in real human serum samples, which proved its practicability., 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 © 2021 Elsevier B.V. All rights reserved.)

It is usually assumed that enzymes retain their native structure during catalysis. However, the aggregation and fragmentation of proteins can be difficult to detect and sometimes conclusions are drawn based on the assumption that the protein is in its native form. We have examined three model enzymes, alkaline phosphatase (AkP), hexokinase (HK) and glucose oxidase (GOx). We find that these enzymes aggregate or fragment after addition of chemical species directly related to their catalysis. We used several independent techniques to study this behavior. Specifically, we found that glucose oxidase and hexokinase fragment in the presence of D-glucose but not L-glucose, while hexokinase aggregates in the presence of Mg 2+ ion and either ATP or ADP at low pH. Alkaline phosphatase aggregates in the presence of Zn 2+ ion and inorganic phosphate. The aggregation of hexokinase and alkaline phosphatase does not appear to attenuate their catalytic activity. Our study indicates that specific multimeric structures of native enzymes may not be retained during catalysis and suggests pathways for different enzymes to associate or separate over the course of substrate turnover.

The selective disruption of nutritional supplements and the metabolic routes of cancer cells offer a promising opportunity for more efficient cancer therapeutics. Herein, a biomimetic cascade polymer nanoreactor (GOx/CAT-NC) was fabricated by encapsulating glucose oxidase (GOx) and catalase (CAT) in a porphyrin polymer nanocapsule for combined starvation and photodynamic anticancer therapy. Internalized by cancer cells, the GOx/CAT-NCs facilitate microenvironmental oxidation by catalyzing endogenous H 2 O 2 to form O 2 , thereby accelerating intracellular glucose catabolism and enhancing cytotoxic singlet oxygen ( 1 O 2 ) production with infrared irradiation. The GOx/CAT-NCs have demonstrated synergistic advantages in long-term starvation therapy and powerful photodynamic therapy (PDT) in cancer treatment, which inhibits tumor cells at more than twice the rate of starvation therapy alone. The biomimetic polymer nanoreactor will further contribute to the advancement of complementary modes of spatiotemporal control of cancer therapy.

Multimodal nanotherapeutic cancer treatments are widely studied but are often limited by their costly and complex syntheses that are not easily scaled up. Herein, a simple formulation of glucose-oxidase-coated CuS nanoparticles was demonstrated to be highly effective for melanoma treatment, acting through a synergistic combination of glucose starvation, photothermal therapy, and synergistic advanced chemodynamic therapy enabled by near-infrared irradiation coupled with Fenton-like reactions that were enhanced by endogenous chloride.

A rapid method for colorimetric monitoring of bacterial viability is described. The colorimetric method was carried out based on glucose oxidase-encapsulated Zn/Co-infinite coordination polymer (Zn/Co-ICP@GOx), which was prepared in aqueous solution free of toxic organic solvents at room temperature. The Zn/Co-ICP@GOx was confirmed to be a robust sphere structure with an average diameter of 147.53 ± 20.40 nm. It integrated the catalytic activity of natural enzyme (GOx) and mimetic peroxidase (Co (П)) all in one, efficiently acting as a biocatalytic cascade platform for glucose catalytic reaction. Exhibiting good multi-enzyme catalytic activity, stability, and selectivity, Zn/Co-ICP@GOx can be used for colorimetric glucose detection. The linear range was 0.01-1.0 mmol/L, and the limit of detection (LOD) was 0.005 mmol/L. As the glucose metabolism is a common expression of bacteria, the remaining glucose can indirectly represent the bacterial viability. Hence, a Zn/Co-ICP@GOx-based colorimetric method was developed for monitoring of bacterial viability. The color was intuitively observed with the naked eye, and the bacterial viability was accurately quantified by measurement of the absorbance at 510 nm. The method was applied to determination of bacterial viability in water and milk samples with recoveries of 99.0-103% and RSD of 0.43-7.5%. The method was rapid (less than 40 min) and applicable to different bacterial species irrespective of Gram-positive and Gram-negative bacteria, providing a universal and promising strategy for real-time monitoring of bacterial viability., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Artificial cell-like communities participate in diverse modes of chemical interaction but exhibit minimal interfacing with their local environment. Here we develop an interactive microsystem based on the immobilization of a population of enzyme-active semipermeable proteinosomes within a helical hydrogel filament to implement signal-induced movement. We attach large single-polynucleotide/peptide microcapsules at one or both ends of the helical protocell filament to produce free-standing soft microactuators that sense and process chemical signals to perform mechanical work. Different modes of translocation are achieved by synergistic or antagonistic enzyme reactions located within the helical connector or inside the attached microcapsule loads. Mounting the microactuators on a ratchet-like surface produces a directional push-pull movement. Our methodology opens up a route to protocell-based chemical systems capable of utilizing mechanical work and provides a step towards the engineering of soft microscale objects with increased levels of operational autonomy., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

The in situ gelation of injectable precursors is desirable in the field of tissue regeneration, especially in the context of irregular defect filling. The current driving forces for fast gelation include the phase-transition of thermally sensitive copolymers, click chemical reactions with tissue components, and metal coordination effect. However, the rapid formation of tough hydrogels remains a challenge. Inspired by aerobic metabolism, we herein propose a tissue-fluid-triggered cascade enzymatic polymerization process catalyzed by glucose oxidase and ferrous glycinate for the ultrafast gelation of acryloylated chondroitin sulfates and acrylamides. The highly efficient production of carbon radicals and macromolecules contribute to rapid polymerization for soft tissue augmentation in bone defects. The copolymer hydrogel demonstrated the regeneration-promoting capacity of cartilage. As the first example of using artificial enzyme complexes for in situ polymerization, this work offers a biomimetic approach to the design of strength-adjustable hydrogels for bio-implanting and bio-printing applications., (© 2021 Wiley-VCH GmbH.)

Nanozyme has been regarded as one of the antibacterial agents to kill bacteria via a Fenton-like reaction in the presence of H 2 O 2 . However, it still suffers drawbacks such as insufficient catalytic activity in near-neutral conditions and the requirement of high H 2 O 2 levels, which would minimize the side effects to healthy tissues. Herein, a mesoporous ceria hollow sphere/enzyme nanoreactor is constructed by loading glucose oxidase in the mesoporous ceria hollow sphere nanozyme. Due to the mesoporous framework, large internal voids, and high specific surface area, the obtained nanoreactor can effectively convert the nontoxic glucose into highly toxic hydroxyl radicals via a cascade catalytic reaction. Moreover, the generated glucose acid can decrease the localized pH value, further boosting the peroxidase-like catalytic performance of mesoporous ceria. The generated hydroxyl radicals could damage severely the cell structure of the bacteria and prevent biofilm formation. Moreover, the in vivo experiments demonstrate that the nanoreactor can efficiently eliminate 99.9% of bacteria in the wound tissues and prevent persistent inflammation without damage to normal tissues in mice. This work provides a rational design of a nanoreactor with enhanced catalytic activity, which can covert glucose to hydroxyl radicals and exhibits potential applications in antibacterial therapy.

Enzymes have demonstrated great potential in the development of advanced electroanalysis devices due to their unique recognition and catalytic properties. However, unsatisfactory stability and limited electron communication of traditional enzyme sensors seriously hinder their large-scale application. In this work, a simple and effective method is proposed to improve the stability of enzyme sensors by using sodium hyaluronate (SH) as a protective film, MXene-Ti 3 C 2 /Glucose oxidase (GOD) as the reaction layer, and chitosan (CS) /reduced graphene oxide (rGO) as the adhesion layer. Results demonstrate that the repeatability of the designed sensor increased by 73.3% after improving the adhesion between the reaction layer and the current collector and that its response ability was greatly enhanced. Moreover, the long-term stability of the electrode surface with SH protective film proved to be superior than that without protective film, which suggests that this design can effectively improve the overall performance of the enzyme biosensor. This work proposed a multi-tier synergistic approach for improving the reliability of enzyme sensors. Graphical abstract Our proposed protective and adhesion layer can greatly improve the stability of enzyme sensor and realize the rapid detection of glucose in serum sample., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

H 2 S and H 2 O 2 are two redox regulating molecules that play important roles in many physiological and pathological processes. While each of them has distinct biosynthetic pathways and signaling mechanisms, the crosstalk between these two species is also known to cause critical biological responses such as protein S-persulfidation. So far, many chemical tools for the studies of H 2 S and H 2 O 2 have been developed, such as the donors and sensors for H 2 S and H 2 O 2 . However, these tools are normally targeting single species (e.g., only H 2 S or only H 2 O 2 ). As such, the crosstalk and synergetic effects between H 2 S and H 2 O 2 have hardly been studied with those tools. In this work, we report a unique H 2 S/H 2 O 2 dual donor system by employing 1-thio-β-d-glucose and glucose oxidase (GOx) as the substrates. This enzymatic system can simultaneously produce H 2 S and H 2 O 2 in a slow and controllable fashion, without generating any bio-unfriendly byproducts. This system was demonstrated to cause efficient S-persulfidation on proteins. In addition, we expanded the system to thiolactose and thioglucose-disulfide; therefore, additional factors (β-galactosidase and cellular reductants) could be introduced to further control the release of H 2 S/H 2 O 2 . This dual release system should be useful for future research on H 2 S and H 2 O 2 .

In this work, cucurbiturils (CBs), a class of macrocyclic supramolecules, were observed to have an interesting peroxidase-like activity, which is metal-free, substrate-specific, thermophilic, acidophilic, and insensitive to ionic strength. By coating CBs on enzyme-encapsulated zeolitic imidazolate framework-8 (ZIF-8), a composite nanozyme was constructed, which retains the catalytic ability of CBs and enzymes and makes them cascade. On addition of the substrate, i.e., the detection target, a highly efficient cascade catalysis can be launched in all the spatial directions to generate sensitive and visible signals. Convenient detection of glucose and cholesterol as models is thereby achieved. More importantly, we have also successfully constructed a composite nanozyme-based sensor array (6 × 8 wells) and thereby achieved simultaneous colorimetric analysis of multiple samples. The concept and successful practice of the construction of the unique core-shell supramolecule/biomolecule@nanomaterial architecture provide the possibility to fabricate next-generation multifunctional materials and create new applications by integrating their unique functions.