Enzyme Mimetic Activity Research Articles

Emerging Roles of Nanozymes in Plant and Environmental Sectors.

The demand for food has increased dramatically as the global population increases, putting more strain on the sustainability of agriculture. To fulfill this requirement, it is imperative to develop brand-new technologies. The application potential of nanozymes in the plant and environmental sectors is progressively becoming apparent as a result of their effective enzymatic catalytic activity and the distinctive characteristics of nanomaterials, including size, specific surface area, optical properties, and thermal properties. Herein, we systematically analyze the catalytic mechanisms of nanozymes with different enzyme-mimetic activities and summarize their applications in improving crop yields by regulating ROS levels and enhancing stress resistance and detecting and removing hazardous pollutants. Finally, we thoroughly analyze the challenges faced by nanozymes regarding size, design, application, economy, and biosafety and look forward to their future development directions to better serve sustainable agriculture.

Nanoscale mineral as a novel class enzyme mimic (mineralzyme) with total antioxidant capacity detection: Colorimetric and smartphone-based approaches

Nanozymes, synthetic nano-scale materials with enzyme-like behavior, have shown remarkable advancements and widespread utilization across various applications. However, the majority of nanozymes require precursor of synthetic-chemicals, which are sometimes expensive and undergo complicated preparations and tedious purification procedures. Therefore, it is of utmost significance to find an enzyme mimic that is affordable, abundant, highly efficient, and sustainable for various applications in biomedicine, environmental sciences, and the food industry. We prove the efficient peroxidase-like activities of the earthly available mineral, barunite-II. The braunite-II mineral micro-nanoparticles (NB) were prepared via physical milling. The enzyme mimetic activity of mineral nanoparticles, referred to as “mineralzyme,” could oxidize the chromogenic blue color of TMB (3,3′,5,5′-tetramethylbenzidine) to oxTMB (3,3′,5,5′-tetramethylbenzidine oxide). Michaelis-Menten constant (Km) and maximum velocity (Vmax) were 135 mM and 62.73 mM min−1 for TMB as a substrate, 139.2 mM, and 2.69 mM min−1 for H2O2 as a substrate. The Km values are much lower than those for HRP. We accurately quantified the total antioxidant capacity in seminal fluid samples from infertile patients using the peroxidase activity of the mineral nanoparticles. This investigation will open new avenues to explore the realm of mineralzyme, revealing its significant potential for a wide range of applications involving diverse enzymatic behaviors.

Enzyme Mimetic Activity of Intercalated Pd–H Nanosheets for Bioanalysis

Enzyme Mimetic Activity of Intercalated Pd–H Nanosheets for Bioanalysis

Enhancing reliability for AFB1 analysis in food: Ratiometric fluorescence/colorimetric dual-modal analysis platform using multifunctional GO-Fe3O4

Adsorption of DNA fluorescent probes on GO-Fe3O4 is a promising strategy for establishing fluorescent bioassays, often using magnetic separation or fluorescence quenching to generate signals. However, there is a lack of systematic understanding of ssDNA-regulated changes in the enzyme-mimetic activity of GO-Fe3O4, and the accuracy of the results of single-mode fluorescence analysis is susceptible to environmental interference. These limit the rational design and scope of application of the methods. Herein, the force and the catalytic mechanism of ssDNA/GO-Fe3O4 interactions were explored in detail. On this basis, a ratiometric fluorescence/colorimetric dual-modal analysis platform was constructed based on the superparamagnetism and DNA controllable peroxidase-like activity of GO-Fe3O4. The ratiometric fluorescent signal was generated by combining 7-amino-4-methyl-3-coumarinylacetic acid (AMCA) labeled aptamer (AMCA-aptamer) with AT hairpin-synthesized copper nanoparticles, which has built-in correction and resistance to environmental interference. The aptamer-modulated peroxidase-like activity of GO-Fe3O4 generated the colorimetric signal. Two signals correct each other to further enhance the reliability of the results. The analytical platform performed satisfactorily for AFB1 detection in the range of 0.1–150 μg/L, and was successfully applied to real samples (peanut, milk powder, and wheat flour). With the support of ImageJ software, quantitative detection was achieved by RGB channel analysis for real-color images, which provides a potential pathway for the rapid detection of food safety.

Ultrasmall CuMn-His Nanozymes with Multienzyme Activity at Neutral pH: Construction of a Colorimetric Sensing Array for Biothiol Detection and Disease Identification.

Biothiol assays offer vital insights into health assessment and facilitate the early detection of potential health issues, thereby enabling timely and effective interventions. In this study, we developed ultrasmall CuMn-Histidine (His) nanozymes with multiple enzymatic activities. CuMn-His enhanced peroxidase (POD)-like activity at neutral pH was achieved through hydrogen bonding and electrostatic effects. In addition, CuMn-His possesses laccase (LAC)-like and superoxide dismutase (SOD)-like activities at neutral pH. Based on three different enzyme mimetic activities of CuMn-His at neutral pH, the colorimetric sensing array without changing the buffer solution was successfully constructed. The array was successfully used for the identification of three biothiols, glutathione (GSH), cysteine (Cys), and homocysteine (Hcy). Subsequently, excellent application results were shown in complex serum and cellular level analyses. This study provides an innovative strategy for the development of ultrasmall bimetallic nanozymes with multiple enzymatic activities and the construction of colorimetric sensing arrays.

Bimetallic Fe3O4@Co3O4/CN as a Nanozyme with Dual Enzyme-Mimic Activities for the Colorimetric Determination of Mercury(II)

Colorimetric biosensor-based nanozymes have received considerable attention in various fields thanks to the advantages of the simple preparation, good stability, and regulable catalytic activity of nanozymes. In this study, a bimetallic nanozyme Fe3O4@Co3O4/CN was prepared via the high-temperature calcination of Fe3O4-PVP@ZIF-67. The material retained its skeletal structure before calcination, which prevented the aggregation of nanoparticles and exposed more active sites of the nanozyme, substantially enhancing the intrinsic dual enzyme-mimetic activities, including peroxidase- and oxidase-like activities. In particular, Fe3O4@Co3O4/CN with oxidase-like activity catalyzed the colorless tetramethylbenzidine (TMB) to become blue oxTMB with oxygen. Reducing glutathione (GSH) could inhibit the above oxidation reaction. In contrast, with respect to the existence of mercury(II), GSH bound to mercury(II) due to the strong affinity between mercury(II) and -SH, thus eliminating the inhibition and restoring the oxTMB signal. A simple and effective colorimetric sensor was fabricated to detect mercury(II) based on the above principles. The proposed measurement had a linear range of 0.1–15 μM and a limit of detection (LOD) of 0.017 μM. It was shown that the established colorimetric sensing system could be successfully applied to detect mercury(II) in water samples, and the Fe3O4@Co3O4/CN nanozyme proved to be a promising candidate for biosensing application.

Reactive oxide species and ultrasound dual-responsive bilayer microneedle array for in-situ sequential therapy of acute myocardial infarction

Acute myocardial infarction (AMI) resulting from coronary artery occlusion stands as the predominant cause of cardiovascular disability and mortality worldwide. An all-encompassing treatment strategy targeting pathological processes of oxidative stress, inflammation, proliferation and fibrotic remodeling post-AMI is anticipated to enhance therapeutic outcomes. Herein, an up-down-structured bilayer microneedle (Ce-CLMs-BMN) with reactive oxygen species (ROS) and ultrasound (US) dual-responsiveness is proposed for AMI in-situ sequential therapy. The upper-layer microneedle is formulated by crosslinking ROS-sensitive linker with polyvinyl alcohol loaded with cerium dioxide nanoparticles (CeNPs) featuring versatile enzyme-mimetic activities. During AMI acute phase, prompted by ischemia-induced microenvironmental redox imbalance, this layer swiftly releases CeNPs, which aid in eliminating excessive ROS and catalyzing oxygen gas (O2) production through multiple enzymatic pathways, thereby alleviating oxidative stress-induced damage and modulating inflammation. In AMI chronic repair phase, micro-nano reactors (CLMs) situated in the lower-layer microneedle undergo cascade reactions with the assistance of US irradiation to generate nitric oxide (NO). As a bioactive molecule with pro-angiogenic and anti-fibrotic effects, NO expedites cardiac repair while attenuating adverse remodeling. Additionally, its antiplatelet-aggregating properties contribute to thromboprophylaxis. In-vitro and in-vivo results substantiate the efficacy of this integrated healing approach in AMI management, showcasing promising prospects for advancing infarcted heart repair.

Redox dual-responsive diaryl ditelluride-containing nanoparticles as peroxidase mimetics

Polymeric nanoparticles with peroxidase-like enzyme mimetic activity were developed by incorporating catalytically active tellurium (Te) functionalities into cross-linked block copolymers. Bis(2-aminophenyl) ditelluride was used as a cross-linker to react with a diblock copolymer bearing pendent activated ester groups to generate of core-cross-linked nanoparticles (DiTe-NP) containing diaryl ditelluride bonds. DiTe-NP nanoparticles exhibit dual redox-responsiveness inherited from the sensitivity of ditelluride to both oxidants and reductants. When DiTe-NP nanoparticles were treated with H2O2, the oxidation of the ditellurides triggered the cleavage of Te-Te bonds to form Te(IV)-associated species. The oxidation-responsiveness endows DiTe-NP nanoparticles with haloperoxidase (HPO)-mimic activity. With NaBr and hydrogen peroxide, they catalyzed the oxidative bromination of phenol red, bromolactonization of 4-pentenoic acid, and bromination of 1,3,5-trimethoxybenzene due to the in-situ generation of hypobromous acid (HOBr) for halogenations. These ditelluride-containing nanoparticles also exhibited glutathione peroxidase (GPx)-like activity and accelerated the reduction of hydroperoxide by a thiol substrate. The cleavage of ditelluride bonds by a thiol substrate leads to the formation of key intermediates in the catalytic cycle of DiTe-NP nanoparticles. DiTe-NP nanoparticles showed enhanced activity for the reduction of hydrophobic cumene peroxide by 4-nitrothiophenol. These dual redox-responsive ditelluride-containing nanoparticles are promising candidates as peroxidase mimetics to apply as catalysts for green halogenation reactions and decompositions of toxic peroxides.

A Macrophage Membrane-Coated Cu-WO3-x-Hydro820 Nanoreactor for Treatment and Photoacoustic/Fluorescence Dual-Mode Imaging of Inflamed Liver Tissue.

A disease-targeting nanoplatform that integrates imaging with therapeutic activity would facilitate early diagnosis, treatment, and therapeutic monitoring. To this end, a macrophage membrane-coated Cu-WO3-x-Hydro820 (CWHM) nanoreactor was prepared. This reactor was shown to target inflammatory tissues. The reactive oxygen species (ROS) such as H2O2 and ·OH in inflammatory tissues can react with Hydro820 in the reactor to form the NIR fluorophore IR820. This process allowed photoacoustic/fluorescence dual-mode imaging of H2O2 and ·OH, and it is expected to permit visual diagnosis of inflammatory diseases. The Cu-WO3-x nanoparticles within the nanoreactor shown catalase and superoxide enzyme mimetic activity, allowing the nanoreactor to catalyze the decomposition of H2O2 and ·O2- in inflammatory cells of hepatic tissues in a mouse model of liver injury, thus alleviating the oxidative stress of damaged liver tissue. This nanoreactor illustrates a new strategy for the diagnosis and treatment of hepatitis and inflammatory liver injury.

Tailoring the Enzyme-Mimetic Activity and Surface Nanostructure of Metal-Phenolic Platform for Colorimetric Detection of l-Cysteine

Tailoring the Enzyme-Mimetic Activity and Surface Nanostructure of Metal-Phenolic Platform for Colorimetric Detection of <scp>l</scp>-Cysteine

Recent Progress and Prospect of Metal–Organic Framework-Based Nanozymes in Biomedical Application

A nanozyme is a nanoscale material having enzyme-like properties. It exhibits several superior properties, including low preparation cost, robust catalytic activity, and long-term storage at ambient temperatures. Moreover, high stability enables repetitive use in multiple catalytic reactions. Hence, it is considered a potential replacement for natural enzymes. Enormous research interest in nanozymes in the past two decades has made it imperative to look for better enzyme-mimicking materials for biomedical applications. Given this, research on metal–organic frameworks (MOFs) as a potential nanozyme material has gained momentum. MOFs are advanced hybrid materials made of inorganic metal ions and organic ligands. Their distinct composition, adaptable pore size, structural diversity, and ease in the tunability of physicochemical properties enable MOFs to mimic enzyme-like activities and act as promising nanozyme candidates. This review aims to discuss recent advances in the development of MOF-based nanozymes (MOF-NZs) and highlight their applications in the field of biomedicine. Firstly, different enzyme-mimetic activities exhibited by MOFs are discussed, and insights are given into various strategies to achieve them. Modification and functionalization strategies are deliberated to obtain MOF-NZs with enhanced catalytic activity. Subsequently, applications of MOF-NZs in the biosensing and therapeutics domain are discussed. Finally, the review is concluded by giving insights into the challenges encountered with MOF-NZs and possible directions to overcome them in the future. With this review, we aim to encourage consolidated efforts across enzyme engineering, nanotechnology, materials science, and biomedicine disciplines to inspire exciting innovations in this emerging yet promising field.

Advances in the Application of Transition-Metal Composite Nanozymes in the Field of Biomedicine.

Due to the limitation that natural peroxidase enzymes can only function in relatively mild environments, nanozymes have expanded the application of enzymology in the biological field by dint of their ability to maintain catalytic oxidative activity in relatively harsh environments. At the same time, the development of new and highly efficient composite nanozymes has been a challenge due to the limitations of monometallic particles in applications and the inherently poor enzyme-mimetic activity of composite nanozymes. The inherent enzyme-mimicking activity is due to Au, Ag, and Pt, along with other transition metals. Moreover, the nanomaterials exhibit excellent enzyme-mimicking activity when composited with other materials. Therefore, this paper focuses on composite nanozymes with simulated peroxidase activity that have been prepared using noble metals such as Au, Ag, and Pt and other transition metal nanoparticles in recent years. Their simulated enzymatic activity is utilized for biomedical applications such as glucose detection, cancer cell detection and tumor treatment, and antibacterial applications.

CuCo2S4 nanoparticles synthesized via a thermal decomposition approach: evaluation of their potential as peroxidase mimics.

The current study demonstrates the synthesis of CuCo2S4 nanoparticles using a novel thermal decomposition approach. The CuCo2S4 nanoparticles were synthesized under various conditions by changing the source of sulfur and the solvent. The CuCo2S4 nanoparticles were characterized using an array of analytical techniques. Powder XRD results indicate the successful formation of CuCo2S4 nanoparticles. TEM results show agglomerated nanoparticles with close to spherical morphology and XPS measurements indicate the presence of Cu2+, Cu+, Co3+, Co2+, and S2- in the samples. The CuCo2S4 nanoparticles exhibit weak ferromagnetic and paramagnetic behaviour at 5 K and 300 K, respectively. The CuCo2S4 nanoparticles were explored for their enzyme mimetic activity using 3,3',5,5' tetramethylbenzidine (TMB) as a substrate. They exhibit better catalytic activity compared to that of a natural enzyme (horseradish peroxidase) and other metal sulfide nanoparticles reported in the literature.

A Dual-Antioxidative Coating on Transmucosal Component of Implant to Repair Connective Tissue Barrier for Treatment of Peri-Implantitis.

Since the microgap between implant and surrounding connective tissue creates the pass for pathogen invasion, sustained pathological stimuli can accelerate macrophage-mediated inflammation, therefore affecting peri-implant tissue regeneration and aggravate peri-implantitis. As the transmucosal component of implant, the abutment therefore needs to be biofunctionalized to repair the gingival barrier. Here, a mussel-bioinspired implant abutment coating containing tannic acid (TA), cerium and minocycline (TA-Ce-Mino) is reported. TA provides pyrogallol and catechol groups to promote cell adherence. Besides, Ce3+ /Ce4+ conversion exhibits enzyme-mimetic activity to remove reactive oxygen species while generating O2 , therefore promoting anti-inflammatory M2 macrophage polarization to help create a regenerative environment. Minocycline isinvolved on the TA surface to create local drug storage for responsiveantibiosis. Moreover, the underlying therapeutic mechanism is revealed whereby the coating exhibits exogenous antioxidation from the inherent properties of Ce and TA and endogenous antioxidation through mitochondrial homeostasis maintenance and antioxidases promotion. In addition, it stimulates integrin to activate PI3K/Akt and RhoA/ROCK pathways to enhance VEGF-mediated angiogenesis and tissue regeneration. Combining the antibiosis and multidimensional orchestration, TA-Ce-Mino repairs soft tissue barriers and effector cell differentiation, thereby isolating the immune microenvironment from pathogen invasion. Consequently, this study provides critical insight into the design and biological mechanism of abutment surface modification to prevent peri-implantitis.

Bioactivity of selenium-containing pyridinium salts: Prospecting future pharmaceutical constituents to treat liver diseases involving oxidative stress.

Redox imbalance leads to oxidative stress that causes irreversible cellular damage. The incorporation of the antioxidant element selenium (Se) in the structure of pyridinium salts has been used as a strategy in chemical synthesis and can be useful in drug development. We investigated the antioxidant activity of Se-containing pyridinium salts (named Compounds 3A, 3B, and 3C) through in vitro tests. We focused our study on liver protein carbonylation, liver lipoperoxidation, free radical scavenging activity (1,1-diphenyl-2-picryl-hydrazil [DPPH]; 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid [ABTS]), and enzyme-mimetic activity assays (glutathione S-transferase [GST]-like; superoxide dismutase [SOD]-like). In addition, 2-(4-chlorophenyl)-2-oxoethyl)-2-((phenylselanyl)methyl)pyridin-1-ium bromide (3C) was selected to evaluate the acute oral toxicity in mice due to the best antioxidant profile. The three compounds were effective in reducing the levels of protein carbonylation and lipoperoxidation in the liver in a µM concentration range. All compounds demonstrated scavenger activity of DPPH and ABTS radicals, and GST-like action. No significant effects were detected in the SOD-like assay. Experimental data also showed that the acute oral treatment of mice with Compound 3C (50 and 300 mg/kg) did not cause mortality or change markers of liver and kidney functions. In summary, our findings reveal the antioxidant potential of Se-containing pyridinium salts in liver tissue, which could be related to their radical scavenging ability and mimetic action on the GST enzyme. They also demonstrate a low toxicity potential for Compound 3C. Together, the promising results open space for future studies on the therapeutic application of these molecules.

Achieving smartphone-based colorimetric assay for Hg2+ with a bimetallic site strategy based on Hg2+-triggered oxidase-like catalytic activity of NSC/Co6Ni3S8 nanocomposite

Modulation of the nanozyme's catalytic activity is crucial for its real applications in detecting target analytes. Herein, we fabricated the nanocomposite (NSC/Co6Ni3S8) of N, S co-doped carbon and Co6Ni3S8 by a facile sol-gel approach. Compared to NSC/Ni9S8, NSC/Co6Ni3S8 with bimetallic active sites displayed better enzyme-mimetic activity. This nanocomposite could catalyze O2 to form ·O2− and oxidize colorless 3, 3′, 5, 5′-tetramethylbenzidine (TMB) into blue oxTMB. The other two free radicals (h+ and ·OH) played minor roles during the catalytic reaction. Hg2+ could integrate with S2− to form HgS and the surface charges of O2 were transferred to Hg2+ to promote O2 adsorption. DFT theoretical calculations highlight that the main reasons for the enhancing effect of Hg2+ on color development results from electron transfer and increased adsorption energy of O2 molecules onto the surface of NSC/Co6Ni3S8. By employing the oxidase-like activity of NSC/Co6Ni3S8 and Hg2+-triggered promoting effect, a colorimetric sensing platform was established for Hg2+ assay with a linear range of 10–200 μg/L and detection limit of 3 μg/L. Through integration with a smartphone-based APP “Thing Identify” software, a visual colorimetric assay for Hg2+ was constructed with a detection limit of 5 μg/L. Compared to the data detected by the mercury vapor meter, the relative recoveries of 92.4–108.1% evidenced the higher accuracy of this smartphone-based visual detection. Overall, the NSC/Co6Ni3S8-based colorimetric assay is convenient, rapid, and visual, and can be applied for routine monitoring of Hg2+ in real-world waters under outdoor conditions.

Bioactive NIR-II gold clusters for three-dimensional imaging and acute inflammation inhibition.

Strong fluorescence and high catalytic activities cannot be achieved simultaneously due to conflicts in free electron utilization, resulting in a lack of bioactivity of most near-infrared-II (NIR-II) fluorophores. To circumvent this challenge, we developed atomically precise Au22 clusters with strong NIR-II fluorescence ranging from 950 to 1300 nm exhibiting potent enzyme-mimetic activities through atomic engineering to create active Cu single-atom sites. The developed Au21Cu1 clusters show 18-fold higher antioxidant, 90-fold higher catalase-like, and 3-fold higher superoxide dismutase-like activities than Au22 clusters, with negligible fluorescence loss. Doping with single Cu atoms decreases the bandgap from 1.33 to 1.28 eV by predominant contributions from Cu d states, and Cu with lost electron states effectuates high catalytic activities. The renal clearable clusters can monitor cisplatin-induced renal injury in the 20- to 120-minute window and visualize it in three dimensions using NIR-II light-sheet microscopy. Furthermore, the clusters inhibit oxidative stress and inflammation in the cisplatin-treated mouse model, particularly in the kidneys and brain.

A Nitro Functionalized MOF with Multi-Enzyme Mimetic Activities for the Colorimetric Sensing of Glucose at Neutral pH.

Benefiting from the advantages like large surface area, flexible constitution, and diverse structure, metal-organic frameworks (MOFs) have been one of the most ideal candidates for nanozymes. In this study, a nitro-functionalized MOF, namely NO2-MIL-53(Cu), was synthesized. Multi-enzyme mimetic activities were discovered on this MOF, including peroxidase-like, oxidase-like, and laccase-like activity. Compared to the non-functional counterpart (MIL-53(Cu)), NO2-MIL-53(Cu) displayed superior enzyme mimetic activities, indicating a positive role of the nitro group in the MOF. Subsequently, the effects of reaction conditions on enzyme mimetic activities were investigated. Remarkably, NO2-MIL-53(Cu) exhibited excellent peroxidase-like activity even at neutral pH. Based on this finding, a simple colorimetric sensing platform was developed for the detection of H2O2 and glucose, respectively. The detection liner range for H2O2 is 1-800 μM with a detection limit of 0.69 μM. The detection liner range for glucose is linear range 0.5-300 μM with a detection limit of 2.6 μM. Therefore, this work not only provides an applicable colorimetric platform for glucose detection in a physiological environment, but also offers guidance for the rational design of efficient nanozymes with multi-enzyme mimetic activities.

Multi-enzyme mimetic iridium nanozymes-based thrombus microenvironment-modulated nanoplatform for enhanced thrombolytic therapy

Intravenous injection of thrombolytic drugs is the most feasible strategy for the clinical treatment of thrombotic diseases, however, its applications are yet limited by narrow therapeutic index, short half-life, and hemorrhagic risk. Inspired by the excessive ROS and inflammatory factors at the thrombus site, we manufactured a thrombus microenvironment-modulated nanosystem, which achieved the targeted delivery of thrombolytic drugs and ROS scavenging agents. Ir monomers (Ir-PVP) with enzyme-mimetic activity were prepared and then wrapped with lumbrokinase (LK) by HA materials of thrombo-inflammatory affinity, forming multifunctional thrombolytic agents Ir-LK@HA. Ir-LK@HA could prolong the biological half-life of LK and target the thrombus site to release LK with reduced bleeding risk. Moreover, Ir-NPs, as both CT imaging contrast agent and ROS scavenger, not only allowed us to observe the progression of thrombolytic treatment through CT imaging in real time, but also played an important role in elimination of excessive ROS at thrombus site. Taken together, the in vitro and in vivo experiments illustrated that Ir-LK@HA with CT function displayed an excellent efficiency in thrombolysis, anti-inflammation and ROS scavenging. Our work may encourage further exploration of thrombus microenvironment-modulated theranostic platforms.

Au/CeO2 NR Restricted Inside Cu-MOFs: A Three-in-One Artificial Enzyme with Synergistically Enhanced Peroxidase-Like Activity for Dual-Mode Sensing of Multiple Biomarkers

Nanomaterials with inherent enzyme-mimetic properties have been extensively studied in biosensing field, but it is still challenging to overcome their natively weak enzyme-mimetic activity and further promote their stabilities. In this work, a three-in-one artificial enzyme with synergistically enhanced peroxidase-like activity and outstanding stability is constructed by restricting Au-loaded CeO2 nanorods (NR) (1% Au:CeO2 mass ratio) inside Cu-based metal–organic frameworks (Cu(PABA)), termed Au1/CeO2 NR@Cu(PABA). It has been demonstrated that the artificial enzyme possesses significantly improved performance in catalytic decomposition of H2O2, with ∼2.41 and ∼1.49 times higher activity than its individual constituents (Au1/CeO2 NR and Cu(PABA)), benefiting from the synergistic effect and the unique restricted structure; owns strong affinities to substrates, with separately ∼8.21- and ∼3.13-fold lower Michaelis constants toward H2O2 and o-phenylenediamine, respectively, than horseradish peroxidase; and exhibits desirable pH (4–12), thermal (30–80 °C), organic solvent (N,N-dimethylformamide, acetone, methanol, etc.), and long-term storage (30 days) stabilities, advantaging practical applications. Taking aforementioned superior properties of Au1/CeO2 NR@Cu(PABA), a universal colorimetric/fluorometric dual-mode sensing platform is built and utilized for detection of biomarkers (e.g., glucose, galactose, and cholesterol) in biological fluids with satisfactory recoveries (90.2–108%). This work offers new horizons in designing high-efficiency, stable, and credible biomimetic catalysts to accelerate future advanced engineering of nanozymes.