Selective Organ-Targeting Hafnium Oxide Nanoparticles with Multienzyme-Mimetic Activities Attenuate Radiation-Induced Tissue Damage.

Radioprotective agents hold clinical promises to counteract off-target adverse effects of radiation and benefit radiotherapeutic outcomes, yet the inability to control drug transport in human organs poses a leading limitation. Based upon a validated rank-based multigene signature model, radiosensitivity indices are evaluated of diverse normal organs as a genomic predictor of radiation susceptibility. Selective ORgan-Targeting (SORT) hafnium oxide nanoparticles (HfO2 NPs) are rationally designed via modulated synthesis by α-lactalbumin, homing to top vulnerable organs. HfO2 NPs like Hensify are commonly radioenhancers, but SORT HfO2 NPs exhibit surprising radioprotective effects dictated by unfolded ligands and Hf(0)/Hf(IV) redox couples. Still, the X-ray attenuation patterns allow radiological confirmation in target organs by dual-beam spectral computed tomography. SORT HfO2 NPs present potent antioxidant activities, catalytically scavenge reactive oxygen species, and mimic multienzyme catalytic activities. Consequently, SORT NPs rescue radiation-induced DNA damage in mouse and rabbit models and provide survival benefits upon lethal exposures. In addition to inhibiting radiation-induced mitochondrial apoptosis, SORT NPs impede DNA damage and inflammation by attenuating activated FoxO, Hippo, TNF, and MAPK interactive cascades. A universal methodology is proposed to reverse radioenhancers into radioprotectors. SORT radioprotective agents with image guidance are envisioned as compelling in personalized shielding from radiation deposition.

Carbon-Based Enzyme Mimetics for Electrochemical Biosensing.

Natural enzymes are used as special reagents for the preparation of electrochemical (bio)sensors due to their ability to catalyze processes, improving the selectivity of detection. However, some drawbacks, such as denaturation in harsh experimental conditions and their rapid de- gradation, as well as the high cost and difficulties in recycling them, restrict their practical applications. Nowadays, the use of artificial enzymes, mostly based on nanomaterials, mimicking the functions of natural products, has been growing. These so-called nanozymes present several advantages over natural enzymes, such as enhanced stability, low cost, easy production, and rapid activity. These outstanding features are responsible for their widespread use in areas such as catalysis, energy, imaging, sensing, or biomedicine. These materials can be divided into two main groups: metal and carbon-based nanozymes. The latter provides additional advantages compared to metal nanozymes, i.e., stable and tuneable activity and good biocompatibility, mimicking enzyme activities such as those of peroxidase, catalase, oxidase, superoxide dismutase, nuclease, or phosphatase. In this review article, we have focused on the use of carbon-based nanozymes for the preparation of electrochemical (bio)sensors. The main features of the most recent applications have been revised and illustrated with examples selected from the literature over the last four years (since 2020).

Elaborating E/Z-Geometry of Alkenes via Cage-Confined Arylation Catalysis of Terminal Olefins.

A visible light photosensitizing metal-organic cage is applied as an artificial supramolecular reactor to control the reaction of aryl radicals with terminal olefins under green light/solvent conditions, which facilitates selective transformation in the confined enzyme-mimicking environment to give a series of geometrically defined E/Z-alkenes. The hydrophobic cage displays good host-guest inclusion with aromatic substrates, promoting Meerwein arylation and protecting E-isomeric products during reaction; while a small amount of benzonitrile can turn on efficient E→Z isomerization. Besides π-π stacking, the hydrogen bonding and halogen bonding interactions also act as control forces for the arylation of aliphatic terminal olefins known as poor acceptors in classic Meerwein arylation. The application of this switchable cage-confined arylation catalysis has been demonstrated by the syntheses of Tapinarof and a marine natural product from the same substrate via controllable E/Z selectivity.

Controlled Self-Assembly of the Catalytic Core of Hydrolases Using DNA Scaffolds.

Precisely organizing functional molecules of the catalytic cores in natural enzymes to promote catalytic performance is a challenging goal in respect to artificial enzyme construction. In this work, we report a DNA-scaffolded mimicry of the catalytic cores of hydrolases, which showed a controllable and hierarchical acceleration of the hydrolysis of fluorescein diacetate (FDA). The results revealed that the efficiency of hydrolysis was greatly increased by the DNA-scaffold-induced proximity of catalytic amino acid residues (histidine and arginine) with up to 4-fold improvement relative to the free amino acids. In addition, DNA-scaffolded one-dimensional and two-dimensional assemblies of multiple catalytic cores could further accelerate the hydrolysis. This work demonstrated that the DNA-guided assembly could be used as a promising platform to build enzyme mimics in a programmable and hierarchical way.

Structural Analysis and Intrinsic Enzyme Mimicking Activities of Ligand-Free PtAg Nanoalloys.

Nanozymes are nanomaterials with biocatalytic properties under physiological conditions and are one class of artificial enzymes to overcome the high cost and low stability of natural enzymes. However, surface ligands on nanomaterials will decrease the catalytic activity of the nanozymes by blocking the active sites. To address this limitation, ligand-free PtAg nanoclusters (NCs) are synthesized and applied as nanozymes for various enzyme-mimicking reactions. By taking advantage of the mutual interaction of zeolitic imidazolate frameworks (ZIF-8) and Pt precursors, a good dispersion of PtAg bimetal NCs with a diameter of 1.78 ± 0.1 nm is achieved with ZIF-8 as a template. The incorporation of PtAgNCs in the voids of ZIF-8 is confirmed with structural analysis using the atomic pair-distribution function and powder X-ray diffraction. Importantly, the PtAgNCs present good catalytic activity for various enzyme-mimicking reactions, including peroxidase-/catalase- and oxidase-like reactions. Further, this work compares the catalytic activity between PtAg NCs and PtAg nanoparticles with different compositions and finds that these two nanozymes present a converse dependency of Ag-loading on their activity. This study contributes to the field of nanozymes and presents a potential option to prepare ligand-free bimetal biocatalysts with sizes in the nanocluster regime.

Nanomaterial Constructs for Catalytic Applications in Biomedicine: Nanobiocatalysts and Nanozymes.

Nanomaterials possess superior advantages due to their special geometries, higher surface area, and unique mechanical, optical, and physicochemical properties. Their characteristics make them great contributors to the development of many technological and industrial sectors. Therefore, novel nanomaterials have an increasing interest in many research areas including biomedicine such as chronic inflammations, disease detection, drug delivery, and infections treatment. Their relevant role is, in many cases, associated with an effective catalytic application, either as a pure catalyst (acting as a nanozyme) or as a support for catalytically active materials (forming nanobiocatalysts). In this review, we analyze the construction of nanozymes and nanobiocatalyst by different existing forms of nanomaterials including carbon-based nanomaterials, metal-based nanomaterials, and polymer-based nanocomposites. Then, we examine successful examples of such nanomaterials employed in biomedical research. The role played by nanomaterials in catalytic applications is analyzed to identify possible research directions toward the development of the field and the achievement of real practicability.

Engineering Antioxidative Cascade Metal-Phenolic Nanozymes for Alleviating Oxidative Stress during Extracorporeal Blood Purification.

Oxidative stress is a compelling risk factor in chronic kidney diseases and is further aggravated for individuals during extracorporeal blood purification, ultimately leading to multiple complications. Herein, antioxidative cascade metal-phenolic nanozymes (metal-tannic acid nanozymes, M-TA NMs) are synthesized via metal ions-mediated oxidative coupling of polyphenols; then M-TA NMs engineered hemoperfusion microspheres (Cu-TAn@PMS) are constructed for alleviating oxidative stress. M-TA NMs show adjustable broad-spectrum antioxidative activities toward multiple reactive nitrogen and oxygen species (RNOS) due to the adjustable catalytic active centers. Importantly, M-TA NMs could mimic the cascade processes of superoxide dismutase and catalase to maintain intracellular redox balance. Detailed structural and spectral analyses reveal that the existence of a transition metal could decrease the electronic energy band gaps of M-TA NMs to offer better electron transfers for RNOS scavenging. Notably, dynamic blood experiments demonstrate that Cu-TAn@PMS could serve as an antioxidant defense system for blood in hemoperfusion to scavenge intracellular reactive oxygen species (ROS) effectively even in the complex blood environment and further protect endogenous antioxidative enzymes and molecules. In general, this work developed antioxidative cascade nanozymes engineered microspheres with excellent therapeutic efficacy for the treatment of oxidative stress-related diseases, which exhibited potential for clinical blood purification and extended the biomedical applications of nanozymes.

Plasmonic Nanozymes: Leveraging Localized Surface Plasmon Resonance to Boost the Enzyme-Mimicking Activity of Nanomaterials.

Nanozymes, a type of nanomaterials that function similarly to natural enzymes, receive extensive attention in biomedical fields. However, the widespread applications of nanozymes are greatly plagued by their unsatisfactory enzyme-mimicking activity. Localized surface plasmon resonance (LSPR), a nanoscale physical phenomenon described as the collective oscillation of surface free electrons in plasmonic nanoparticles under light irradiation, offers a robust universal paradigm to boost the catalytic performance of nanozymes. Plasmonic nanozymes (PNzymes) with elevated enzyme-mimicking activity by leveraging LSPR, emerge and provide unprecedented opportunities for biocatalysis. In this review, the physical mechanisms behind PNzymes are thoroughly revealed including near-field enhancement, hot carriers, and the photothermal effect. The rational design and applications of PNzymes in biosensing, cancer therapy, and bacterial infections elimination are systematically introduced. Current challenges and further perspectives of PNzymes are also summarized and discussed to stimulate their clinical translation. It is hoped that this review can attract more researchers to further advance the promising field of PNzymes and open up a new avenue for optimizing the enzyme-mimicking activity of nanozymes to create superior nanocatalysts for biomedical applications.

Effective fabrication of cellulose nanofibrils supported Pd nanoparticles as a novel nanozyme with peroxidase and oxidase-like activities for efficient dye degradation.

Nanozyme-based dye degradation methods are promising for the remediation of water pollution. Though Pd nanoparticles (PdNPs) are known to act as nanozymes, their dye degradation capability has not been investigated. Low nanozyme activities, easy aggregation, difficulties in recovery and reuse are the major challenges in achieving this. For the first time, cellulose nanofibrils-supported PdNPs (PdNPs/PCNF) as a novel nanozyme with good peroxidase and oxidase-mimicking activities and easy recyclability is explored for dye degradation. An efficient and rapid method of PdNPs/PCNF preparation was demonstrated by adjusting the pH and microwave irradiation. Enzyme kinetic studies revealed good kinetic parameters and specific activities of 415 and 277 U/g for peroxidase and oxidase, respectively. PdNPs/PCNF offered 99.64% degradation of methylene blue within 12 min (0.468 min-1) with 0.4 M H2O2 at pH 5.0. Mechanistic studies revealed the involvement of hydroxyl and superoxide radicals. Owing to the network-like structure of PCNF, films and foams were prepared, their dye degradation potentials were compared, and recyclability was tested. Successful degradation of mixed dye solutions and spiked real water samples was achieved and a continuous flow method was demonstrated using a foam-packed column.

The steadfast Au@Pt soldier: Peroxide-tolerant nanozyme for signal enhancement in lateral flow immunoassay of peroxidase-containing samples.

We report the approach for the detection of Au@Pt core@shell nanoparticles (nanozymes) with peroxidase-mimicking activity (PMA) in samples with high endogenous peroxidase activity (EPA). Unlike the endogenous peroxidases in plant extracts that are inhibited by elevated H2O2 (>20 mM), the PMA of nanozymes was stable in concentrated H2O2 (up to 4 M). Such a different stability of enzymes and Au@Pt to the substrate allowed for eliminating EPA and detecting only nanozymes. The developed approach was used for reaching a lower limit of detection (LOD) and eliminating the background for the lateral flow immunoassay (LFIA) of the important plant pathogen potato virus X (PVX) in leaf and tuber extracts. Using the PMA of Au@Pt, the LOD was reduced to 4 and 8 pg/mL in tuber and leaf extracts, respectively. The LOD values are 250- and 500-times lower in comparison with LFIA with conventional gold nanoparticles. The developed approach of peroxidases inhibition is universal for bioanalytical methods, and its applicability was confirmed by the elimination of EPA in three matrixes (serum, potato leaf and tuber extracts).

High Carbonization Temperature to Trigger Enzyme Mimicking Activities of Silk-Derived Nanosheets.

Herein, it is demonstrated that N-rich carbonized silk fibroin materials (CSFs) can serve as efficient peroxidase, and oxidase mimics. Their enzyme-like activities are highly dependent on carbonization conditions. CSFs obtained at low temperatures do not exhibit significant catalytic reactivity, while their enzyme-like catalysis performance is greatly activated after high-temperature treatment. Such a phenomenon is mainly ascribed to the increase of graphitization degree and graphitic nitrogen and the emergence of disordered graphitic structures during the formation of turbostratic carbon. In addition, inspired by the excellent photothermal conversion efficiency, and temperature-dependent catalytic behavior of CSFs, near-infrared light can be used to remotely control their enzyme-like activities. More importantly, as-prepared robust silk-derived nanosheets can be applied to photothermal-catalytic cancer therapy and sensing. It is believed that such a smart artificial enzyme system will throw up exciting new opportunities for the chemical industry and biotechnology.

Sesquiterpene Cyclizations inside the Hexameric Resorcinarene Capsule: Total Synthesis of δ-Selinene and Mechanistic Studies.

The synthesis of terpene natural products remains a challenging task due to the enormous structural diversity in this class of compounds. Synthetic catalysts are unable to reproduce the tail-to-head terpene cyclization of cyclase enzymes, which create this diversity from just a few simple linear terpene substrates. Recently, supramolecular structures have emerged as promising enzyme mimetics. In the present study, the hexameric resorcinarene capsule was utilized as an artificial cyclase to catalyze the cyclization of sesquiterpenes. With the cyclization reaction as the key step, the first total synthesis of the sesquiterpene natural product δ-selinene was achieved. This represents the first total synthesis of a sesquiterpene natural product that is based on the cyclization of a linear terpene precursor inside a supramolecular catalyst. To elucidate the reaction mechanism, detailed kinetic studies and kinetic isotope measurements were performed. Surprisingly, the obtained kinetic data indicated that a rate-limiting encapsulation step is operational in the cyclization of sesquiterpenes.

Colorimetric enzymatic determination of glucose based on etching of gold nanorods by iodine and using carbon quantum dots as peroxidase mimics.

Carbon quantum dots (CQDs) with peroxidase-mimicking activity were successfully prepared from litchi rind. A colorimetric method for glucose determination was developed based on etching of gold nanorods (GNRs) using CQDs as peroxidase mimetic. The glucose oxidase-catalyzed oxidation of glucose leads to the generation of H2O2 which oxidizes added iodide under formation of elemental iodine under the catalytic action of CQDs. Iodine then etches the GNRs along the longitudinal direction due to the higher reaction activities at the tips of GNRs. This results in a stepwise decrease in the maximum absorption wavelength of the GNRs, from initially 953 nm to finally 645 nm. Under the optimized conditions, the shift in the maximum absorption wavelength decreases linearly in the 0.01-2.0 mM glucose concentration range, and the detection limit is 3.0 µM. Importantly, this method was applied to the determination of glucose in human serum. It is perceived that the CQDs are valuable peroxidase mimics due to their ease of preparation, low costs and stable catalytic activity. Graphical abstract Carbon quantum dots were prepared from litchi rind. They can induce the oxidation of gold nanorods in the presence of I- ions and H2O2. This finding was applied to design a colorimetric assay for glucose.

Vinyl Triflimides-A Case of Assisted Vinyl Cation Formation.

A new concept for selectivity control in carbocation-driven reactions has been identified which allows for the chemo-, regio-, and stereoselective addition of nucleophiles to alkynes-assisted vinyl cation formation-enabled by a Li+ -based supramolecular framework. Mechanistic analysis of a model complex (Li2 NTf2 + ⋅3 H2 O) confirms that solely the formation of a complex between the incoming nucleophile and the transition state of the alkyne protonation is responsible for the resulting selective N addition to the vinyl cation. Into the bargain, a general, operationally simple synthetic procedure to previously inaccessible vinyl triflimides is provided.

Colorimetric zearalenone assay based on the use of an aptamer and of gold nanoparticles with peroxidase-like activity.

An aptamer based colorimetric assay is described for the determination of zearalenone (ZEN). It is based on the inhibition of the peroxidase-mimicking activity of gold nanoparticles (AuNPs) by the ZEN aptamer. However, in the presence of ZEN, the aptamer is bound by ZEN and can no longer inhibit the peroxidase-mimicking activity of AuNPs. The color change of solution is related to ZEN concentration and observed with bare eyes. Under optimal conditions, the absorbance (at 630 nm) increases linearly in the ZEN concentration range of 10-250 ng·mL-1, and the limit of detection is 10 ng·mL-1. The specificity of the assay was verified by studying the effect of potential interferents. The recoveries from ZEN spiked corn and corn oil range from 92 to 110%, and the relative standard deviations are between 2.4 and 6.4%. The results are in good agreement with those obtained by an ELISA. Graphical abstract Schematic presentation of colorimetric assay for rapid and sensitive determination of zearalenone (ZEN) based on the inhibition of ZEN aptamer on the the peroxidase-like activity of gold nanoparticle (AuNPs).

A highly selective colorimetric sulfide assay based on the inhibition of the peroxidase-like activity of copper nanoclusters.

The authors report that sulfide ions are capable of inhibiting the peroxidase-like activity of copper nanoclusters (CuNCs). The catalytic activity of CuNCs toward the oxidation of the chromogenic substrate 3,3',5,5'-tetramethylbenzidine by H2O2 is remarkably decreased in the presence of sulfide. Based on this finding, a colorimetric assay was developed for the rapid determination of sulfide. Best operated at a wavelength of 652 nm, it has a 0.5 µM detection limit. The method is highly selective and has been successfully applied to the quantification of sulfide in environmental water samples. Graphical abstract The catalytic activity of CuNCs toward the oxidation of 3,3',5,5'-tetramethylbenzidine by H2O2 is remarkably decreased in the presence of sulfide ions. This finding has been applied to design a method for colorimetric quantification of sulfide ions in environmental samples.

Self-Assembly of Enzyme-Like Nanofibrous G-Molecular Hydrogel for Printed Flexible Electrochemical Sensors.

Conducting hydrogels provide great potential for creating designer shape-morphing architectures for biomedical applications owing to their unique solid-liquid interface and ease of processability. Here, a novel nanofibrous hydrogel with significant enzyme-like activity that can be used as "ink" to print flexible electrochemical devices is developed. The nanofibrous hydrogel is self-assembled from guanosine (G) and KB(OH)4 with simultaneous incorporation of hemin into the G-quartet scaffold, giving rise to significant enzyme-like activity. The rapid switching between the sol and gel states responsive to shear stress enables free-form fabrication of different patterns. Furthermore, the replication of the G-quartet wires into a conductive matrix by in situ catalytic deposition of polyaniline on nanofibers is demonstrated, which can be directly printed into a flexible electrochemical electrode. By loading glucose oxidase into this novel hydrogel, a flexible glucose biosensor is developed. This study sheds new light on developing artificial enzymes with new functionalities and on fabrication of flexible bioelectronics.

Biomimetic nitrogen doped titania nanoparticles as a colorimetric platform for hydrogen peroxide detection.

Nanoparticles proved a viable alternative to the already used sensing and diagnostics methods due to their low cost, good stability, easy availability and easy synthesis. In the present approach, nitrogen doped titania nanoparticles are prepared through freeze drying method, and subsequently stabilized through ionic liquid. These nanoparticles were characterized through various techniques such as X-ray diffraction (XRD), Fourier transformation infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), BET pore size and surface area analyzer, X-ray photoelectron spectroscopy (XPS) and UV-Visible diffuse reflectance spectroscopy (UV-Vis. DRS). The synthesized nitrogen doped titania nanoparticles were proved to be a novel peroxidase mimetic with great potential to catalyze oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide (H2O2) to form a blue color product. As a proof of concept, this new enzyme mimic was used as a robust nanoprobe for the detection of hydrogen peroxide with improved analytical characteristics. A linear response for hydrogen peroxide detection was obtained in the range of 10-300µmol/L, with a detection limit of 2.5µmol/L. Taking into account the valuable intrinsic peroxidase activity, the present work may find widespread applications in the field of sensors and biosensors for diverse applications.

Nanocomposite incorporating V2O5 nanowires and gold nanoparticles for mimicking an enzyme cascade reaction and its application in the detection of biomolecules.

Artificial enzyme mimics are a current research interest, and many nanomaterials have been found to display enzyme-mimicking activity. However, to the best of our knowledge, there have not hitherto been any reports on the use of pure nanomaterials to construct a system capable of mimicking an enzyme cascade reaction. Herein, we describe the construction of a novel nanocomposite consisting of V2O5 nanowires and gold nanoparticles (AuNPs) through a simple and facile chemical method, in which V2O5 and AuNPs possess intrinsic peroxidase and glucose oxidase (GOx)-like activity, respectively. Results suggest that this material can mimic the enzyme cascade reaction of horseradish peroxidase (HRP) and GOx. Based on this mechanism, a direct and selective colorimetric method for the detection of glucose has been successfully designed. Because single-strand and double-strand DNA (ssDNA and dsDNA) have different deactivating effects on the GOx-like activity of AuNPs, the sensing of target complementary DNA can also be realized and disease-associated single-nucleotide polymorphism of DNA can be easily distinguished. Our study opens a new avenue for the use of nanomaterials in enzyme mimetics, and holds promise for the further exploration of nanomaterials in creating alternative catalytic systems to natural enzymes.

A minimalist chemical model of matrix metalloproteinases--can small peptides mimic the more rigid metal binding sites of proteins?

In order to mimic the active center of matrix metalloproteinases (MMPs), we synthesized a pentadecapeptide (Ac-KAHEFGHSLGLDHSK-NH2) corresponding to the catalytic zinc(II) binding site of human MMP-13. The multi-domain structural organization of MMPs fundamentally determines their metal binding affinity, catalytic activity and selectivity. Our potentiometric, UV-visible, CD, EPR, NMR, mass spectrometric and kinetic studies are aimed to explore the usefulness of such flexible peptides to mimic the more rigid metal binding sites of proteins, to examine the intrinsic metal binding properties of this naked sequence, as well as to contribute to the development of a minimalist, peptide-based chemical model of MMPs, including the catalytic properties. Since the multiimidazole environment is also characteristic for copper(II), and recently copper(II) containing variants of MMPs have been identified, we also studied the copper(II) complexes of the above peptide. Around pH 6-7 the peptide, similarly to MMPs, offers a {3Nim} coordination binding site for both zinc(II) and copper(II). In the case of copper(II), the formation of amide coordinated species at higher pH abolished the analogy with the copper(II) containing MMP variant. On the other hand, the zinc(II)-peptide system mimics some basic features of the MMP active sites: the main species around pH7 (ZnH2L) possesses a {3Nim,H2O} coordination environment, the deprotonation of the zinc-bound water takes place near the physiological pH, it forms relatively stable ternary complexes with hydroxamic acids, and the species ZnH2L(OH) and ZnH2L(OH)2 have notable hydrolytic activity between pH7 and 9.