Mechanistic and kinetic insights into size-dependent activity in ultra-small Pt/CNTs nanozymes during antibacterial process

In enzyme-like catalytic reactions, the size effect of nanoparticles has been an essential yet unclear factor for the catalytic activity of nanozymes. Moreover, the synthesis of nanozymes with controllable size and electronic structures represents a grand challenge, which limits the systemic exploration the underlying nature of their structure–property relations and practical application. In this work, we proposed a novel strategy to regulate the size of Pt (0.55 ～ 2.81 nm) by atomic layer deposition for precisely tailoring Pt-based nanozymes. The size-dependent electronic and kinetic effects have been observed for the peroxidase-like reaction and antibacterial process, revealing a volcano-type dependence of intrinsic activity on Pt nanoparticle sizes, and the optimum Pt nanoparticle size was found to be ca. 1.69 nm. A combination of kinetic study and XPS analyses, as well as multiple nanozyme characterizations, demonstrates that Pt nanoparticles with an appropriate size contribute to proper affinity to the substrates, relating to a high ratio of Pt⁰/Pt²⁺ on the surface of Pt nanoparticles, which is beneficial to obtain the excellent catalytic performance and antibacterial activity. Our work provides insights for an in-depth understanding size-dependent catalytic mechanism of nanozymes during antibacterial processes.     

Unveiling the Intrinsic Catalytic Activities of Single‐Gold‐Nanoparticle‐Based Enzyme Mimetics

Gold nanoparticles (AuNPs) have been demonstrated to serve as effective nanomaterial‐based enzyme mimetics (nanozymes) for a number of enzymatic reactions under mild conditions. The intrinsic glucose oxidase and peroxidase activities of single AuNPs and Ag–Au nanohybrids, respectively, were investigated by single NP collision electrochemical measurements. A significantly high turnover number of nanozymes was obtained from individual catalytic events compared with the results from the classical, ensemble‐averaged measurements. The unusual enhancement of catalytic activity of single nanozymes is believed to originate from the high accessible surface area of monodispersed NPs and the high activities of carbon‐supported NPs during single‐particle collision at a carbon ultramicroelectrode. This work introduces a new method for the precise characterization of the intrinsic catalytic activities of nanozymes, giving further insights to the design of high‐efficiency nanomaterial catalysts.     

Molecularly Imprinted Nanozymes with Free Substrate Access for Catalyzing the Ligation of ssDNA Sequences

Nanozymes have attracted wide attention for the unique advantages of low cost, high stability and designability. Molecularly imprinted polymers (MIPs) have demonstrated great potential as a new type of nanozymes due to their excellent specificity and high affinity. However, effective approaches for creating molecularly imprinted nanozymes still remain limited. Herein, reverse microemulsion template docking surface imprinting (RMTD-SI) is reported as a new approach for the rational design and engineering of nanozymes with free substrate access for the ligation of ssDNA sequences. As a proof of the principle, octa-deoxyribonucleotide-imprinted nanoparticles were successfully prepared. Using tetradeoxyribonucleotides and octa-deoxyribonucleotide as substrates, the properties, catalytic activity and behavior of the imprinted nanoparticles were thoroughly investigated. The imprinted nanozyme exhibited an enhanced reaction speed (by up to 41-fold) and good sequence selectivity towards substrate tetra-deoxyribonucleotides. More interestingly, due to the open substrate access, the imprinted nanozyme also allowed the ligation of a ssDNA that fully matched with the imprinted cavity and other ssDNA substrates to form longer sequences, but at the price of substrate selectivity. Thus, this study provides not only a new avenue to the rational design and synthesis of molecularly imprinted nanozymes but also new insights of their catalytic behavior.     

Recent progress in carbon-dots-based nanozymes for chemosensing and biomedical applications

Nanozymes are nanomaterials with enzyme-like activities that efficiently overcome the drawbacks of natural enzymes in biosensing, detection, and biomedical fields, and they are the most widely used artificial enzymes. Owing to their excellent catalytic characteristics, biocompatibility, and environmental favorability, carbon-dots-based (CDs) nanozymes have inspired a research upsurge. However, no review focusing on CDs nanozymes has been published, even though substantial advances have been achieved. Herein, the advances, catalytic activities, and applications of CDs nanozymes are highlighted and summarized. In addition, the critical issues and challenges of researching nanozymes are discussed. We hope that this review will broaden the horizons of nanozymes and CDs nanozymes, as well as promote their development.     

Recent advances in the development of colorimetric analysis and testing based on aggregation-induced nanozymes

Colorimetric sensing strategies as a powerful point-of-care testing(POCT) tool have attracted significant interest in various chem/biosensing applications.Taking the excellent bare-eye-detectable signaling feature,nanozymes-based colorimetric sensors enable more potential applications and have been a new forefront in the colorimetric POCT analysis toward different target analytes.However,the low catalytic activity of nanozymes in most cases limits their practical application.Recent efforts demonstrate that the aggregation-induced nanozymes provide a general means to modulate nanozymes activity and enhance colorimetric sensing performances of some nanozymes-based colorimetric sensors.But there are few reports are explored to discuss and review such aggregation-induced nanozymes and their colorimetric sensing applications.To highlight the advances and progress in aggregation-induced nanozymes based colorimetric assays,we herein summary the fundamentals,classify and applications of this newlydeveloping field,focusing on the aggregation-induced activity enhancement of nanozymes(AIAEnanozymes) with a significant "signal-on" feature and aggregation-induced activity inhibition of nanozymes(AIAI-nanozymes) with a dramatical "signal-of" characteristics.Finally,we also propose the current challenges and the future prospects on both AIAE-nanozymes and AIAI-nanozymes.     

Defect-rich and ultrathin nitrogen-doped carbon nanosheets with enhanced peroxidase-like activity for the detection of urease activity and fluoride ion

Although carbon nanozymes have attracted great interest due to their good biocompatibility, low cost,and high stability, designing high-active carbon nanozymes still faces great challenges. Herein, ultrathin nitrogen-doped carbon nanosheets with rich defects(d-NC) were prepared through a high-temperature annealing process, using potassium chloride and ammonium chloride as templates. Owing to the large specific surface area, rich defects and the high exposure of active sites, the proposed d-NC na...     

Fabrication of a Ratiometric Fluorescence Sensor Based on Carbon Dots as Both Luminophores and Nanozymes for the Sensitive Detection of Hydrogen Peroxide

The construction of novel fluorescent nanozymes is highly desirable for providing new strategies for nanozyme-based sensing systems. Herein, a novel ratiometric fluorescence sensing platform was constructed based on carbon dots (CDs) as both luminophores and nanozymes, which could realize the sensitive detection of hydrogen peroxide (H₂O₂). CDs with peroxidase-mimicking activity were prepared with a one-step hydrothermal method using L-histidine as an inexpensive precursor. CDs had bright blue fluorescence. Due to the pseudo-peroxidase activity, CDs catalyzed the oxidation of o-phenylenediamine (OPD) with H₂O₂ to generate 2,3-diaminophenolazine (DAP). The fluorescence resonance energy transfer (FRET) between CDs and DAP resulted in a decrease in the fluorescence of CDs and an increase in the fluorescence of DAP, leading to a ratiometric fluorescence system. The free radical trapping experiment was used to investigate the reactive oxygen radicals (ROS) in the catalytic process of CD nanozymes. The enzymatic parameters of CD nanozymes, including the Michaelis constant (K_m) and the maximum initial reaction velocities (V_max), were investigated. A good affinity for both OPD and H₂O₂ substrates was proven. Based on the FRET between CDs and OPD, a ratiometric fluorescence analysis of H₂O₂ was achieved and results ranged from 1 to 20 μM and 20 to 200 μM with a low limit of detection (LOD, 0.42 μM). The detection of H₂O₂ in milk was also achieved.     

Advances in Single‐Atom Nanozymes Research†

Nanozymes with intrinsic enzyme‐like properties have attracted significant interest owing to their capability to address the limitations of traditional enzymes such as fragility, high cost and difficult mass production. However, the currently reported nanozymes are generally less active than natural enzymes. In recent years, with the rapid development of nanoscience and nanotechnology, single‐atom nanozymes (SAzymes) with well‐defined electronic and geometric structures have shown a promise to serve as direct surrogates of traditional enzymes by mimicking the highly evolved catalytic center of natural enzymes. In this review, we will introduce the enzymatic characteristics and recent advances of SAzymes, and summarize their significant applications from in vitro detection to in vivo monitoring and therapy.     

Self-Indicative Gold Nanozyme for H2O2 and Glucose Sensing

In addition to superior enzyme-mimicking abilities, nanozymes also have intrinsic physicochemical properties. Integrating the enzyme-like activities and tunable physicochemical properties into a single nanoparticle is a promising strategy for versatile nanozyme design and application. Herein, a composite nanozyme in which Au nanoparticles are encapsulated by Au nanoclusters (AuNP@AuNCs) is presented. By integrating the peroxidase-mimicking ability of fluorescent Au NCs with the glucose oxidase-like activity of Au NPs, the composite nanozyme realized cascade assay of glucose without the aid of external indicators. Compared to traditional multistep colorimetric methods, the analytical process was highly simplified by using the self-responsive nanozyme. This synthetic strategy provided valuable insights into exploring talented nanozymes for sensing diverse targets.     

Liposome-Boosted Peroxidase-Mimicking Nanozymes Breaking the pH Limit

Peroxidase-mimicking nanozymes such as Fe₃O₄ nanoparticles are promising substitutes for natural enzymes like horseradish peroxidase. However, most such nanozymes work efficiently only in acidic conditions. In this work, the influence of various liposomes on nanozyme activity was studied. By introducing negatively charged liposomes, peroxidase-mimicking nanozymes achieved oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in neutral and even alkaline conditions, although the activity towards anionic 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was inhibited. The Fe₃O₄ nanoparticles adsorbed on the liposomes without disrupting membrane integrity as confirmed by fluorescence quenching, dye leakage assays, and cryo-electron microscopy. Stabilization of the blue-colored oxidized products of TMB by electrostatic interactions was believed to be the reason for the enhanced activity. This work has introduced lipids to nanozyme research, and it also has practically important applications for using nanozymes at neutral pH, such as the detection of hydrogen peroxide and glucose.     

Defect‐Rich Adhesive Nanozymes as Efficient Antibiotics for Enhanced Bacterial Inhibition

Nanozymes have emerged as a new generation of antibiotics with exciting broad‐spectrum antimicrobial properties and negligible biotoxicities. However, their antibacterial efficacies are unsatisfactory due to their inability to trap bacteria and their low catalytic activity. Herein, we report nanozymes with rough surfaces and defect‐rich active edges. The rough surface increases bacterial adhesion and the defect‐rich edges exhibit higher intrinsic peroxidase‐like activity compared to pristine nanozymes due to their lower adsorption energies of H₂O₂ and desorption energy of OH*, as well as the larger exothermic process for the whole reaction. This was demonstrated using drug‐resistant Gram‐negative Escherichia coli and Gram‐positive Staphylococcus aureus in vitro and in vivo. This strategy can be used to engineer nanozymes with enhanced antibacterial function and will pave a new way for the development of alternative antibiotics.     

Tuning iron spin states in single-atom nanozymes enables efficient peroxidase mimicking

The large-scale application of nanozymes remains a significant challenge owing to their unsatisfactory catalytic performances. Featuring a unique electronic structure and coordination environment, single-atom nanozymes provide great opportunities to vividly mimic the specific metal catalytic center of natural enzymes and achieve superior enzyme-like activity. In this study, the spin state engineering of Fe single-atom nanozymes (FeNC) is employed to enhance their peroxidase-like activity. Pd nanoclusters (Pd_NC) are introduced into FeNC, whose electron-withdrawing properties rearrange the spin electron occupation in Fe(ii) of FeNC–Pd_NC from low spin to medium spin, facilitating the heterolysis of H₂O₂ and timely desorption of H₂O. The spin-rearranged FeNC–Pd_NC exhibits greater H₂O₂ activation activity and rapid reaction kinetics compared to those of FeNC. As a proof of concept, FeNC–Pd_NC is used in the immunosorbent assay for the colorimetric detection of prostate-specific antigen and achieves an ultralow detection limit of 0.38 pg mL⁻¹. Our spin-state engineering strategy provides a fundamental understanding of the catalytic mechanism of nanozymes and facilitates the design of advanced enzyme mimics.

Spin-state engineering was proposed to enhance the peroxidase-like activity of single-atom nanozymes through the electron-withdrawing properties of Pd nanoclusters, which facilitates the heterolysis process of H₂O₂ and the desorption of H₂O.     

煤基碳点的制备及其在检测痕量Cu2+中的应用

以新疆五彩湾煤炭为原料,通过混酸(H2 SO4+HNO3)/超声结合法制得煤基碳点(C-dots),对其结构进行了表征,考察了其作为荧光探针检测水体中金属离子的性能.结果表明,此煤基C-dots是一类具有sp2碳构筑的多层石墨烯碎片结构环壁,且链接了硝基等含氧基团的稠环芳烃结构粒子,粒径为(8±4)nm,具有良好的吸收和荧光发射能力及水分散性;在痕量Cu2+存在下,此C-dots能够选择性地发生荧光猝灭,对Cu2+的检出限低至9.6 nmol/L,是一种极为有效的Cu2+荧光检测剂.     

Rhodium-Based Nanozymes: Recent Advances and Challenges

Rhodium (Rh) is a non-toxic transition metal used as various nanomaterials with unique structures and properties. Rh-based nanozymes can mimic the activities of natural enzymes, overcome the limitation of the application scope of natural enzymes, and interact with various biological microenvironments to play a variety of functions. Rh-based nanozymes can be synthesized in various ways, and different modification and regulation methods can also enable users to control catalytic performance by adjusting enzyme active sites. The construction of Rh-based nanozymes has attracted great interest in the biomedical field and impacted the industry and other areas. This paper reviews the typical synthesis and modification strategies, unique properties, applications, challenges, and prospects of Rh-based nanozymes. Next, the unique features of Rh-based nanozymes are emphasized, including adjustable enzyme-like activity, stability, and biocompatibility. In addition, we discuss Rh-based nanozymes biosensors and detection, biomedical therapy, and industrial and other applications. Finally, the future challenges and prospects of Rh-based nanozymes are proposed.     

Spectroscopic and Isothermal Titration Calorimetry Studies of Binding Interactions Between Carbon Nanodots and Serum Albumins

Carbon nanodots (C-dots) have attracted great attention as a new class of luminescent nanomaterials. In order to better understand the basic behavior of C-dots in biological systems, the binding characteristics of C-dots with bovine serum albumin (BSA) and human serum albumin (HSA) were investigated using spectroscopic approaches and isothermal titration calorimetry at pH 7.4. We found that the intrinsic fluorescence of BSA and HSA was quenched by the C-dots with a dynamic quenching mode. It was proved that the C-dots had little influence on the conformation of BSA and HSA by their UV–vis and circular dichroism spectra. Some important thermodynamic parameters were calculated, and the positive values of ΔH° and ΔS° indicate that the binding process was endothermic, and that the interaction was driven by favorable entropy and unfavorable enthalpy. It also showed that the hydrophobic force played a major role in the binding process.     

Nitrogen and boron co-doped graphene nanoribbons as peroxidase-mimicking nanozymes for enhanced biosensing

The rational design of nanozymes with superior activities is essential for improving bioassay performances. Herein, nitrogen and boron co-doped graphene nanoribbons(NB-GNRs) are prepared by a hydrothermal method using urea as the nitrogen source and boric acid as the boron source, respectively.The introduction of co-doped and edge structures provides high defects and active sites. The resultant NB-GNRs nanozymes show superior peroxidase-like activities to nitrogen-doped and boron-doped counterpa...     

Single-Atom Nanozymes: Fabrication,Characterization, Surface Modification and Applications of ROS Scavenging and Antibacterial

Nanozymes are nanomaterials with intrinsic natural enzyme-like catalytic properties. They have received extensive attention and have the potential to be an alternative to natural enzymes. Increasing the atom utilization rate of active centers in nanozymes has gradually become a concern of scientists. As the limit of designing nanozymes at the atomic level, single-atom nanozymes (SAzymes) have become the research frontier of the biomedical field recently because of their high atom utilization, well-defined active centers, and good natural enzyme mimicry. In this review, we first introduce the preparation of SAzymes through pyrolysis and defect engineering with regulated activity, then the characterization and surface modification methods of SAzymes are introduced. The possible influences of surface modification on the activity of SAzymes are discussed. Furthermore, we summarize the applications of SAzymes in the biomedical fields, especially in those of reactive oxygen species (ROS) scavenging and antibacterial. Finally, the challenges and opportunities of SAzymes are summarized and prospected.     

超分子手性聚苯胺-金复合纳米酶的构建及其对映选择性催化

对映选择性催化是生物体内普遍存在的反应,与生命的产生代谢有着非常紧密的联系。设计研发具有高对映选择性催化效果的纳米酶在各类生物医药相关领域都至关重要。目前,关于纳米酶的研究大都集中在提高其催化活性,而涉及纳米酶对映选择性的研究相对较少。已有对映选择性酶催化报道表明,手性纳米酶主要通过手性分子修饰纳米颗粒来构建。考虑到天然酶的选择性不仅仅取决于氨基酸等手性分子的手性,而且与蛋白质空间排列和折叠所产生的超分子手性微环境密切相关,因此构建具有超分子手性微环境的纳米酶也成为设计具有优异对映选择性纳米酶的有效途径。此外,为了进一步提高手性纳米酶的对映选择性,深入理解手性纳米酶选择性因子的影响因素也成为一个重要研究方向。基于此,本文构建了一种由不含任何手性分子的M-聚苯胺(M-PANI)扭曲纳米带和三种不同尺寸(3、10和16 nm)的金纳米颗粒(AuNPs)组成的超分子纳米复合材料。扫描电子显微镜、透射电子显微镜、紫外-可见吸收光谱和X射线光电子能谱结果证实了M-PANI-Au超分子纳米复合材料的成功制备。同时,圆二色光谱显示M-PANI-Au超分子复合材料具有清晰的手性信号,表明它们在手性纳米催化方面具有潜在应用。以手性R-/S-3, 4-二羟基苯丙氨酸(R-/S-DOPA)对映体的催化氧化为模型反应,该类纳米酶对R-DOPA的催化选择性均高于对S-DOPA。进一步研究表明,得益于超分子手性聚苯胺载体和3 nm AuNPs之间的强手性传递作用,3 nm Au NPs(2.59)负载的M-PANI比10 nm Au NPs(1.46)和16 nm NPs(1.58)负载的M-PANI具有更高的选择因子。这一发现阐明了手性转移是调控对映选择性催化的关键影响因素,为负载型超分子手性纳米酶的构建和设计提供了方向和指导。     

Spectrophotometric investigation of the effect of beta-cyclodextrin on the intramolecular cyclization reaction of catecholamines using rank annihilation factor analysis

The catalytic effect of β-cyclodextrin (β-CD) on the intramolecular cyclization reaction of catecholamines dopamine, methyldopa and levodopa after their oxidation with periodate was investigated spectrophotometrically using rank annihilation factor analysis (RAFA). The method was based on the effect of β-CD concentration on the intramolecular cyclization reaction of the investigated catecholamines after their oxidation with potassium periodate. In order to perform RAFA, concentration profiles were calculated by optimizing the value of formation constant. The rank of original data matrix can be reduced by one by annihilating the information of the cyclization reaction in the absence of β-CD. The rate constant for the reactions in the presence and absence of β-CD was also determined. The best estimation of rate constant in the presence of β-CD reduces the rank of the system to zero (noise level). The method was evaluated using synthetic data as well as experimental data and good results were obtained.     

A Nanozyme with Photo‐Enhanced Dual Enzyme‐Like Activities for Deep Pancreatic Cancer Therapy

Nanozymes have attracted extensive interest owing to their high stability, low cost and easy preparation, especially in the field of cancer therapy. However, the relatively low catalytic activity of nanozymes in the tumor microenvironment (TME) has limited their applications. Herein, we report a novel nanozyme (PtFe@Fe₃O₄) with dual enzyme‐like activities for highly efficient tumor catalytic therapy. PtFe@Fe₃O₄ shows the intrinsic photothermal effect as well as photo‐enhanced peroxidase‐like and catalase‐like activities in the acidic TME, thereby effectively killing tumor cells and overcoming the tumor hypoxia. Importantly, a possible photo‐enhanced synergistic catalytic mechanism of PtFe@Fe₃O₄ was first disclosed. We believe that this work will advance the development of nanozymes in tumor catalytic therapy.