Single-atom Nanozyme Research Articles

Boosting Enzyme-like Activities via Atomization of Cerium for Tumor Microenvironment-Responsive Cascade Therapy.

Nanozyme catalytic therapeutic efficacy is limited by the finite enzyme activity and specificity. In this work, nitrogen-doped carbon loaded with a cerium single-atom nanozyme (Ce SAs@NC) is synthesized, exhibiting tumor specificity and excellent multiple enzyme-like activities. Compared with nitrogen-doped carbon loaded with CeO2 nanoparticles, Ce SAs@NC shows excellent peroxidase-like and catalase-like activity. Ce SAs@NC can convert intracellular hydrogen peroxide into cytotoxic hydroxyl radical and O2, which can be further transferred to superoxide radicals. Cascade enzyme reactions not only alleviate the hypoxic microenvironment of tumors but also induce lipid peroxidation and apoptosis or necrosis of tumor cells. The mild photothermal action will enhance the enzyme-like activities of Ce SAs@NC rather than induce the production of heat shock proteins to protect tumor cells. In addition, Ce SAs@NC can regulate the immune environment, stimulate M1 macrophages to trigger immune responses, and inhibit tumor proliferation. Thanks to the combination of the size effect of the single atoms, photothermal influence, multiple enzyme-like activities, and immunological effect, the Ce SAs@NC platform appears to have tumor specificity, less toxic side effects, and a high curative effect both in vitro and in vivo.

A dual-signal sensing platform based on single atomic iron-nitrogen-carbon nanozyme for the detection of neurodegenerative disease-related bioactive molecules acetylcholinesterase and dopamine

In this work, a novel iron-nitrogen-carbon single-atom nanozyme (Fe-N800 SAzyme) with peroxidase-like activity was synthesized by doping Fe in zeolitic imidazolate frameworks (ZiF-8) and utilizing graphitic C3N4 (g-C3N4) as a nitrogen source in the pyrolysis process. Acetylcholine (ACh) was continuously oxidized by acetylcholinesterase (AChE) and choline oxidase (CHO) to generate H2O2 through an enzymatic cascade reaction. The interaction between Fe-N800 SAzyme and H2O2 led to the generation of reactive oxygen species (ROS) that could then oxidize 3,3’,5,5’-tetramethylbenzidine (TMB) to oxidized TMB (oxTMB), which exhibited an absorption peak at 650 nm. Lysozyme (LZ)-functionalized 5-methyl-2-thiouracil (MTU) gold/silver nanoclusters (Au/Ag/LZ-MTU), which exhibited a fluorescence peak at 610 nm, could be quenched by oxTMB through the inner filter effect (IFE). Dopamine (DA) was able to consume ROS and hinder the oxidation of TMB. Therefore, by monitoring the change of fluorescence at 610 nm and absorption at 650 nm, a dual signal fluorometric and colorimetric sensing platform for AChE and DA was established. The AChE activity was monitored through the variation of fluorescence (I610) or colorimetric (A650) intensity, which had a low limit of detection (LOD) of 0.057 U/L and 0.11 U/L, respectively. The fluorescence and colorimetric methods were also able to sensitively monitor the DA activity with a LOD of 0.83 μM and 2.7 μM, respectively. Finally, the developed method could satisfactorily quantify AChE and DA in human serum, whole blood, and DA hydrochloride injection samples.

A Fe/Zn Dual Single-Atom Nanozyme with High Peroxidase Activities for Detection of Penicillin G.

Penicillin G (PG) is a common antibiotic, and its accumulation in the environment can pose a threat to the ecological system and ultimately impact human health. Nanozymes have emerged as highly stable enzyme mimics that can be utilized as sensors to achieve the sensitive detection of specific antibiotics. Herein, we report on a dual single-atom Fe/Zn nanozyme (DSAzyme) synthesized from Fe-imidazole as the guest and zeolite imidazole framework-8 as the host. The DSAzyme exhibits intriguing properties that mimic the activities of two natural enzymes: peroxidase and lactamase. Both activities are utilized for the design of a colorimetric sensor for the specific detection of PG: the peroxidase activity enables color generation from 3,3',5,5'-tetramethylbenzidine and H2O2, and the lactamase activity provides the recognition of PG. The nanozyme consists of many Fe-N4 and Zn-N4 site and mechanistic characterizations by experimental investigations and theoretical calculations identify Fe-N4 as the main active center for the peroxidase activity and Zn-N4 as the main binding site for PG. The sensor can achieve a limit of detection of 47 nM, is able to detect PG from real-life samples, remains fully functional after 8-month storage, and retain high activities after reuse for fives times. Taken together, our study provides a new approach to the detection of antibiotics in environmental samples.

Smartphone-based colorimetric paper chip sensor using single-atom nanozyme for the detection of carbofuran pesticide residues in vegetables

Carbofuran (CBF), which exhibit high toxicity, persistent residues, ease of accumulation, and resistance to degradation, pose serious threats to human health and harm the ecological environment. Therefore, there is an urgent need to develop a rapid and accurate method for detecting CBF. In this work, a low-cost, portable, and easy-to-use paper chip biosensor was developed, integrating smartphones for the detection of CBF pesticide residues. This biosensor facilitates rapid on-site testing, meeting the needs for immediate analysis. CBF has the ability to inhibit acetylcholinesterase (AChE) activity. In the presence of AChE, acetylthiocholine (ATCh) is hydrolyzed to produce thiocholine (TCh). TCh, in turn, can inhibit the catalytic activity of Ni-N-C single-atom nanozymes (SAzyme) synthesized using Ni(OH)2 nanochip as a metal precursor, which possess high peroxidase activity. Consequently, the concentration of CBF can be determined by observing the resultant color changes. The results showed that this sensor had a good linear response in the range of CBF concentration from 10 to 500 ng/mL, and the LOD was as low as 8.79 ng/mL. In testing three actual samples—Chinese cabbage, cabbage, and lettuce—the recoveries ranged from 81.09% to 125.27%. This demonstrated that the proposed smartphone-based colorimetric paper chip sensor, utilizing Ni-N-C SAzyme, offers an immediate, convenient, and rapid new strategy for detecting CBF.

Single-atom ruthenium nanozyme-induced signal amplification strategy in photoelectrochemical aptasensor for ultrasensitive detection of chloramphenicol

To develop ultrasensitive and rapid antibiotics residue detection method is crucial for ensuring food safety and protecting human health. Herein, a novel photoelectrochemical (PEC) aptasensor integrated with single-atom ruthenium (Ru) nanozyme-mediated catalytic precipitation as a valuable signal amplification strategy, have been established for ultrasensitive chloramphenicol (CAP) detection. Particularly, the exceptional peroxidase-mimicking activity of single-atom Ru nanozyme is responsible for accelerating the oxidation of 4-chloro-1-naphthol (4-CN) to produce insoluble precipitate on the electrode, which in turn causes a notable reduction in the photocurrent. Whereas, when CAP is present, the aptamer is liberated away the electrode because of its potent affinity with CAP, resulting in an elevation of the photocurrent signal, enhancing the detection sensitivity. Importantly, the signal amplification strategy combines the effective photoactive material of Au nanoparticles/CdS quantum dot/TiO2 composites, a PEC aptasensor for determination of CAP with an ultralow detection limit of 4.12 pM is achieved in a self-powered mode with great selectivity and accuracy. This work proposes a novel reasonable approach utilizing high-activity single-atom nanozyme to induce signal amplification strategy for the advancement of single-atom nanozyme in ultrasensitive PEC biosensor, and further creates new avenues for ultrasensitive detection beyond antibiotics residue.

Targeting senescence with radioactive 223Ra/Ba SAzymes enables senolytics-unlocked One‐Two punch strategy to boost anti-tumor immunotherapy

Senescent cells are characterized by a persistent cessation of their cell cycle, rendering them valuable targets for anti-tumor strategies in cancer treatment. Numerous studies have explored induced senescence as a promising approach in tumor therapy. Nevertheless, these treatments often come with drawbacks, including adverse side effects and weaker senescence-inducing effects. To address these challenges, we synthesized 223Ra/Ba single-atom nanozyme (SAzyme), wherein Ba SAzyme acts concurrently as a carrier for 223RaCl2, facilitating targeted delivery and minimizing side effects. The 223Ra/Ba SAzyme complex enhances various enzyme-mimicking functions, including catalase (CAT) and peroxidase (POD) activities. Importantly, 223Ra/Ba SAzyme induces cellular senescence and boost anti-tumor immunity. The persistent presence of a senescence-associated secretory phenotype (SASP) in the tumor microenvironment presents risks of immune suppression and tumor recurrence, which can be effectively mitigated by senolytics. As a result, 223Ra/Ba SAzyme were combined with anti-PD-L1 checkpoint blockade to achieve a one-two punch therapy, wherein 223Ra/Ba SAzyme exploits senescence followed by anti-PD-L1 therapy to eradicate senescent cells. This one‐two punch strategy approach presents a straightforward and potent intervention for both primary tumors and distant tumor.

A single-atom nanozyme-enabled strategy for rapid, visual, and real-time detection of polystyrene nanoplastics in water

Polystyrene nanoplastics (PSNPs) in water environment have become a critical issue to ecosystems and human health due to their small sizes, easier diffusion and higher hazard. It is an urgent need to develop an efficient, rapid, and on-site detection strategy for PSNPs. A single-atom nanozyme of zeolitic imidazolate framework (ZIF-FeSAN) was prepared using hemoglobin as template and Fe-source. ZIF-FeSAN possessed porous structure, a high surface area (994.91 m2/g), and maximumly exposed Fe atoms, resulting in a high peroxidase (POD)-like activity. ZIF-FeSAN exhibited excellent adsorption capability (83.46 mg/g, 10 min) for PSNPs via the electrostatic and π-π interactions. After adsorption of PSNPs, the active centers of Fe atom were shielded with a great decreasing of POD-like activity. Given the ‘on-off’ effect of POD-like activity, a ZIF-FeSAN colorimetric sensor was developed for PSNPs (20, 30, 50, 100, and 150 nm) detection. The limit of detection (LOD) was 0.212, 4.544, 7.624, 16.955, and 24.171 mg/L for 20, 30, 50, 100, and 150 nm, respectively. Meanwhile, a smartphone-assisted ZIF-FeSAN platform was designed to rapid, on-site, and visual detection of PSNPs, and the corresponding LODs were 0.851, 17.564, 34.554, 67.254, and 76.124 mg/L, respectively. The ZIF-FeSAN-based strategies have practical application in water samples due to high satisfactory recoveries (91.47–108.12 %) and good selectivity. This proof-of-concept study paves the way toward the on-site, rapid, and visual quantitative detection of PSNPs in water environment with avoiding the use of large and expensive instruments.

Single-atom nanozyme liposome-integrated microneedles for in situ drug delivery and anti-inflammatory therapy in Parkinson’s disease

Treatment for Parkinson’s disease (PD) has been impeded by inefficient treatment results and multiple membrane barriers during drug delivery. This study reports the design, synthesis, and application of microneedles (MNs) loaded with mitochondrion-targeted liposome encapsulated iron (Fe)-isolated single-atom nanozymes (Mito@Fe-ISAzyme, MFeI), called MFeI MNs, for in situ drug delivery into the brain parenchyma and efficient enrichment of drugs in lesion sites. In in vitro experiments, MFeI can scavenge reactive oxygen species (ROS) and protect the neurons via mitochondrial targeting, guaranteeing the subsequent treatment of PD. Using PD mouse models, we compared the intravenous injection of MFeI with the brain in situ administration of MFeI MNs (in situ MFeI MNs). Results showed that in situ MFeI MNs significantly improved the deep penetration of the drug into brain parenchyma, especially in the vital pathological sites such as the substantia nigra pars compacta and striatum. Importantly, ROS elimination and neuroinflammatory remission in the lesion site were observed, thereby efficiently alleviating the behavioral disorders and pathological symptoms of PD mice. Therefore, the MNs system for in situ single-atom nanozyme liposome delivery exhibits great potential in PD treatment.Graphical abstract

Iron Single-Atom Nanozyme with Inflammation-Suppressing for Inhibiting Multidrug-Resistant Bacterial Infection and Facilitating Wound Healing.

Infection with drug-resistant bacteria and the formation of biofilms are the main factors contributing to wound healing insufficiency. Antibacterial agents with enzyme-like properties have exhibited considerable potential for efficient eradication of drug-resistant microorganisms due to their superior sensitivities and minimal side effects. In this work, we prepared a kind of Fe-centered single-atom nanozyme (Fe-SAzyme) with high biocompatibility and stability via a facile one-pot hydrothermal method, which was suitable for the treatment of wounds infected with drug-resistant bacteria. The Fe-SAzyme exhibited remarkable peroxidase-like catalytic activities, catalyzing the conversion of hydrogen peroxide (H2O2) to highly toxic hydroxyl radicals (•OH), which could not only damage bacterial cells but also inhibit, disrupt, and eradicate the formation of bacterial biofilms. Thus, Fe-SAzyme demonstrated a broad-spectrum antibacterial performance capable of effectively eliminating multidrug-resistant bacteria. The coexistence of ferrous (Fe2+) and ferric (Fe3+) ions in Fe-SAzyme conferred the nanozyme with anti-inflammatory activity, effectively suppressing excessive inflammation. Meanwhile, Fe-SAzyme could significantly downregulate inflammatory cytokines tumor necrosis factor-α and interleukin-1β and upregulate growth factors VEGF and epidermal growth factor, which can prevent bacterial infection, mitigate inflammation, promote fibroblast proliferation, and improve wound closure. Thus, Fe-SAzyme had shown favorable therapeutic efficiency in promoting bacteria-infected wound healing. This study provides Fe-SAzyme as a promising candidate for the development of new strategies to treat multidrug-resistant bacterial infections.

Study on the Electrochemiluminescence Emission Mechanism of HOF-14 and Its Multimode Sensing and Imaging Application.

A novel hydrogen-bonded organic framework (HOF-14) has attracted much attention due to its excellent biocompatibility and low toxicity, but its research in the electrochemiluminescence (ECL) field has not been reported. In this work, the annihilation-type and coreactant-type ECL emission mechanisms of HOF-14 were studied systematically for the first time. It was found that the ECL quantum efficiency of HOF-14/TEA coreactant system was the highest, which was 1.82 times that of Ru(bpy)32+/TEA. Further, the ECL emission intensity of HOF-14/TEA system could achieve colorimetric (CL) imaging of mobile phone. We also discovered that HOF-14 had superior photoelectrochemical (PEC) performance. Based on the above research results, a unique HOF-14-based multimode sensing and imaging platform was constructed. The antibiotic Enrofloxacin (ENR) was selected as the detection target, and the Cu-Zn bimetallic single-atom nanozyme (Cu-Zn/SAzyme) with excellent peroxidase (POD)-like activity was used to prepare quenching probes. When the target ENR was present, Cu-Zn/SAzyme quenching probes were introduced to the surface of HOF-14 by the dual-aptamer sandwich method. Cu-Zn/SAzyme could catalyze diaminobenzidine (DAB) to produce brown precipitations to quench the ECL, PEC, and CL signals of HOF-14, realizing multimode detection of ENR. This work not only discovered excellent ECL and PEC property of new HOF-14 material but also systematically studied the ECL emission mechanism of HOF-14, and proposed a novel multimode sensing and imaging platform, which greatly improved the detection accuracy of target and showed great contributions to the field of ECL analysis.

Spatiotemporally Guided Single-Atom Bionanozyme for Targeted Antibiofilm Treatment.

The heterogeneous and dynamic microenvironment of biofilms complicates bacterial infection treatment. Nanozyme catalytic therapy has recently been promising in treating biofilm infections. However, active nanozymes designed with the required precision targeting the biofilm microenvironment are lacking. This work proposes a spatiotemporally guided single-atom bionanozyme (BioSAzyme) for targeted antibiofilm therapy based on protein engineering of copper single-atom nanozyme (Cu SAzyme). The Cu SAzyme, synthesized via a novel mechanochemistry-assisted method, features highly accessible Cu-N4 active sites exposed on 2D N-doped carbon, exhibiting excellent triple enzyme-like activities according to experimental results and density functional theory calculations. Inheriting biofunctionality from both glucose oxidase and concanavalin A, BioSAzyme can localize the biofilm glycocalyx and catalyze endogenous glucose into H₂O₂ and gluconic acid, thus triggering multiplex cascade reactions with pH self-adaption to consume glucose and glutathione and generate •OH radicals. This spatiotemporally guided bionanocatalytic agent effectively inhibits E. coli O157: H7 and methicillin-resistant S. aureus biofilms in vitro and in vivo. Taking together, this work opens up new avenues for the rational design of single-atom nanozymes for precise antibiofilm therapy.

Biomimetic Single-Atom Nanozyme for Dual Starvation-Enhanced Breast Cancer Immunotherapy.

Cold exposure (CE) therapy is an innovative and cost-efficient cancer treatment that activates brown adipose tissue to compete for glucose uptake, leading to metabolic starvation in tumors. Exploring the combined antitumor mechanisms of CE and traditional therapies (such as nanocatalysis) is exciting and promising. In this study, a platelet membrane biomimetic single-atom nanozyme (SAEs) nanodrug (PFB) carrying bis-2-(5-phenylacetamido-1, 2, 4-thiadiazol-2-yl) ethyl sulfide (BPTES) is developed for use in cancer CE therapy. Owing to the platelet membrane modification, PFB can effectively target tumors. Upon entering cancer cells, the dual starvation effect induced by CE treatment and BPTES can significantly diminish intracellular glucose and ATP levels, resulting in a substantial reduction in cellular (glutathione) GSH, which can enhance the cytotoxic efficacy of reactive oxygen species generated by SAEs. This strategy not only boosts ROS production in tumors, but also strengthens immune responses, particularly by increasing memory T-cell abundance and suppressing distant tumor growth and tumor metastasis. Compared with SAEs therapy alone, this combined approach offers superior benefits for tumor immunotherapy. This study achieves a combination of CE and nanomedicines for the first time, providing new ideas for future nanomedicine combination therapy modalities.

Single-Atom Iridium Nanozyme-Based Persistent Luminescence Nanoparticles for Multimodal Imaging-Guided Combination Tumor Therapy.

Persistent luminescence nanoparticles (PLNPs) can achieve autofluorescence-free afterglow imaging, while near-infrared (NIR) emission realizes deep tissue imaging. Nanozymes integrate the merits of nanomaterials and enzyme-mimicking activities with simple preparation. Here PLNPs are prepared of Zn1.2Ga1.6Ge0.2O4:Cr0.0075 with NIR emission at 700 nm. The PLNPs are then incubated with IrCl3 solution, and the nanoparticles are collected and annealed at 750 °C to obtain iridium@PLNPs. Iridium is observed on the PLNPs at the atomic level as a single-atom nanozyme with peroxidase-like catalytic activity, photothermal conversion, and computed tomography (CT) contrast capability. After coating with exosome membrane (EM), the Ir@PLNPs@EM composite exhibits long-lasting NIR luminescence, peroxidase-like catalytic activity, photothermal conversion, and CT contrast capability, with the targeting capability and biocompatibility from EM. Thus, NIR afterglow/photothermal/CT trimodal imaging-guided photothermal-chemodynamic combination therapy is realized as validated with the in vitro and in vivo inhibition of tumor growth, while toxicity and side effects are avoided as drug-free treatment. This work offers a promising avenue for advanced single-atom nanozyme@PLNPs to promote the development of nanozymes and PLNPs for clinical applications.

Cu–N–C single-atom nanozyme as an ultrasensitive sensing platform for α-glucosidase detection

Alpha-glucosidase (α-Glu) is a hydrolase that promotes the rise of glucose in the blood during the digestion process, and it plays a crucial role in regulating glucose levels within a normal range. Herein, we developed an ultrasensitive colorimetric platform by preparing a Cu–N–C single-atom nanozyme (Cu–N–C SAzyme) with a potent oxidase-like effect on accurately detection of α-Glu and efficient screening of its potential inhibitors. This artificial nanoenzyme was able to catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxide (oxTMB) without the assistance of H2O2. In addition, with the participation of α-Glu, l-ascorbic acid-2-O-α-d-glucopyranose (AAG) can be catalyzed to produce ascorbic acid (AA), which significantly reduces the oxidation, resulting in a noticeable fading of the blue color. Therefore, we designed a straightforward colorimetric test for α-Glu activity, achieving detection limits as low as 0.09 U/L and a linear response from 0.25 to 8 U/L. Most importantly, this platform has been effectively applied to measure α-Glu in real samples.

Inflammation-free electrochemical in vivo sensing of dopamine with atomic-level engineered antioxidative single-atom catalyst

Electrochemical methods with tissue-implantable microelectrodes provide an excellent platform for real-time monitoring the neurochemical dynamics in vivo due to their superior spatiotemporal resolution and high selectivity and sensitivity. Nevertheless, electrode implantation inevitably damages the brain tissue, upregulates reactive oxygen species level, and triggers neuroinflammatory response, resulting in unreliable quantification of neurochemical events. Herein, we report a multifunctional sensing platform for inflammation-free in vivo analysis with atomic-level engineered Fe single-atom catalyst that functions as both single-atom nanozyme with antioxidative activity and electrode material for dopamine oxidation. Through high-temperature pyrolysis and catalytic performance screening, we fabricate a series of Fe single-atom nanozymes with different coordination configurations and find that the Fe single-atom nanozyme with FeN4 exhibits the highest activity toward mimicking catalase and superoxide dismutase as well as eliminating hydroxyl radical, while also featuring high electrode reactivity toward dopamine oxidation. These dual functions endow the single-atom nanozyme-based sensor with anti-inflammatory capabilities, enabling accurate dopamine sensing in living male rat brain. This study provides an avenue for designing inflammation-free electrochemical sensing platforms with atomic-precision engineered single-atom catalysts.

Copper Single-Atom Nanozyme Mimicking Galactose Oxidase with Superior Catalytic Activity and Selectivity.

Due to the low stability and high cost of some natural enzymes, nanozymes have been developed as enzyme-imitating nanomaterials. Single-atom nanozymes are a class of nanozymes with metal centers that mimic the structure of metal-based natural enzymes. Herein, Cu-N-C single-atom nanozyme (SAN) is synthesized with excellent peroxidase- and enhanced oxidase-like activities to mimic the action of natural galactose oxidase. Cu-SAN demonstrates stereospecific activity akin to that of natural galactose oxidase by oxidizing D-galactose and primary alcohol but not L-Galactose or other carbohydrates. The SAN can catalyze the oxidation of galactose in the presence of oxygen, producing hydrogen peroxide as a sub-product. The produced hydrogen peroxide then oxidizes 3,3',5,5'-tetramethylbenzidine catalyzed by the SAN, yielding the typical blue product. The relationship between absorbance and galactose concentration is linear in the 1-60µm range with a detection limit as low as 0.23µm. This strategy can be utilized in the diagnosis of galactosemia disorder and detection of galactose in some dairy and other commercial products. DFT calculations clarify the high activity of the Cu sites in the POD-like reaction and explain the selectivity of the Cu-SAN oxidase-like reaction toward D-galactose.

Atomically Engineered Chlorine Coordination of Iron in Active Centers for Selectively Catalytic H2O2 Decomposition Toward Efficient Antitumor-Specific Therapy.

The intervention of endogenous H2O2 via nanozymes provides a potential antitumor-specific therapy; however, the role of the nanozyme structure in relation to the selective decomposition of H2O2 to hydroxyl radicals (•OH) is yet to be fully understood, which limits the development of this therapeutic approaches. Herein, an iron single-atom nanozyme (Fe─N2Cl2─C SAzyme) is reported, which is prepared through precise Fe─Cl coordination based on the construction of a characteristic Fe-containing molecule. Fe─N2Cl2─C exhibits efficient catalytic H2O2 decomposition (2.19 × 106 mm-1 s-1), which is the highest among reported SAzymes. More importantly, it is found that H2O2 selectively decomposed into •OH on the Fe─N2Cl2─C surface, which is attributable to the d orbitals of the Fe active center matching the O-2p electrons of the adsorbed hydroxide (*OH) intermediate. Fe─N2Cl2─C is strongly cytotoxic toward a variety of cancer-cell lines in vitro but not to normal cells. Furthermore, Fe─N2Cl2─C shows an outstanding specific therapeutic effect in vivo; it efficiently destroys solid malignant tumors without injuring normal tissue. Altogether, these findings highlight the selective catalytic decomposition of H2O2 to •OH, which is achieved by engineering the active center on the atomic level, thereby providing an avenue for the development of specific nanomedicines with efficient antitumor activities.

Atom-pair engineering of single-atom nanozyme for boosting peroxidase-like activity

Constructing atom-pair engineering and improving the activity of metal single-atom nanozyme (SAzyme) is significant but challenging. Herein, we design the atom-pair engineering of Zn-SA/CNCl SAzyme by simultaneously constructing Zn-N4 sites as catalytic sites and Zn-N4Cl1 sites as catalytic regulator. The Zn-N4Cl1 catalytic regulators effectively boost the peroxidase-like activities of Zn-N4 catalytic sites, resulting in a 346-fold, 1496-fold, and 133-fold increase in the maximal reaction velocity, the catalytic constant and the catalytic efficiency, compared to Zn-SA/CN SAzyme without the Zn-N4Cl1 catalytic regulator. The Zn-SA/CNCl SAzyme with excellent peroxidase-like activity effectively inhibits tumor cell growth in vitro and in vivo. The density functional theory (DFT) calculations reveal that the Zn-N4Cl1 catalytic regulators facilitate the adsorption of *H2O2 and re-exposure of Zn-N4 catalytic sites, and thus improve the reaction rate. This work provides a rational and effective strategy for improving the peroxidase-like activity of metal SAzyme by atom-pair engineering.

Carbon dots-supported Zn single atom nanozymes for the catalytic therapy of diabetic wounds

Diabetic wound treatment continues to be a significant clinical issue due to higher levels of oxidative stress, susceptibility to bacterial infections, and chronic inflammatory responses during healing. We rationally developed and synthesized an ultra-small carbon dots (C-dots) loaded with zinc single-atom nanozyme (Zn/C-dots) with the aim of promoting wounds healing by nanocatalytic treatment, especially targeting its complex pathological microenvironment. Zinc single atoms and C-dots form a dual catalytic system with higher enzymatic activity. Furthermore, the Zn/C-dots nanozyme effectively enters cells, accumulates at mitochondria, and removes excess ROS, protecting cells from oxidative stress damage and limiting the release of pro-inflammatory cytokines, hence reducing inflammation. Zinc can synergistically increase the antibacterial action of C-dots (the effective antibacterial rate of 100 µg/mL Zn/C-dots was above 90 %). Unlike traditional C-dots, Zn/C-dots can cause endothelial cell migration and the formation of new blood vessels. In vitro cytotoxicity, blood compatibility, and in vivo toxicity studies of Zn/C-dots show that they are biocompatible. We subsequently utilized the Zn/C-dots nanozymes to treat diabetic rats' chronic wounds for external use, combining them with ROS-responsive hydrogels to create an antioxidative system (H-Zn/C-dots). The hydrogels anchored the Zn/C-dots nanozymes to the wound, allowing for long-term treatment. The results revealed that H-Zn/C-dots can considerably reduce inflammation, accelerate angiogenesis, collagen deposition, and promote tissue remodeling at the diabetic wound site. After 14 days, the wound area had decreased to approximately 9.19 %, making it a potential treatment. Statement of significanceAn ultra-small carbon dot with a zinc single-atom nanozyme was designed and manufactured. Zn/C-dots possess antibacterial, ROS-scavenging, and angiogenesis activities. In vivo, the multifunctional ROS-responsive hydrogel incorporating Zn/C-dots could speed up diabetic wound healing.

Development of a Zn-Based Single-Atom Nanozyme for Efficient Hydrolysis of Glycosidic Bonds.

Hydrolytic enzymes are essential components in second-generation biofuel technology and food fermentation processes. Nanozymes show promise for large-scale industrial applications as replacements for natural enzymes due to their distinct advantages. However, there remains a research gap concerning glycosidase nanozymes. In this study, a Zn-based single-atom nanozyme (ZnN4-900) is developed for efficient glycosidic bond hydrolysis in an aqueous solution. The planar structure of the class-porphyrin N4 material approximatively mimicked the catalytic centers of natural enzymes, facilitating oxidase-like (OXD-like)activity and promoting glycosidic bond cleavage. Theoretical calculations show that the Zn site can act as Lewis acids, attacking the C─O bond in glycosidic bonds. Additionally, ZnN4-900 has the ability to degrade starch and produce reducing sugars that increased yeast cell biomass by 32.86% and ethanol production by 14.56%. This catalyst held promising potential for enhancing processes in ethanol brewing and starch degradation industries.