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Abstract Nanomaterials excel in mimicking the structure and function of natural enzymes while being far more interesting in terms of structural stability, functional versatility, recyclability, and large‐scale preparation. Herein, the story assembles hemin, histidine analogs, and G‐quadruplex DNA in a catalytically competent supramolecular assembly referred to as assembly‐activated hemin enzyme (AA‐heminzyme). The catalytic properties of AA‐heminzyme are investigated both in silico (by molecular docking and quantum chemical calculations) and in vitro (notably through a systematic comparison with its natural counterpart horseradish peroxidase, HRP). It is found that this artificial system is not only as efficient as HRP to oxidize various substrates (with a turnover number kcat of 115 s−1) but also more practically convenient (displaying better thermal stability, recoverability, and editability) and more economically viable, with a catalytic cost amounting to

Abstract In enzymatic catalysis, the redox site and Lewis acid are the two main roles played by metal to assist amino acids. However, the reported enzyme mimics only focus on the redox-active metal as redox site, while the redox-inert metal as Lewis acid has, to the best of our knowledge, not been studied, presenting a bottleneck of enzyme mimics construction. Based on this, a series of highly efficient MxV2O5·nH2O peroxidase mimics with vanadium as redox site and alkaline-earth metal ion (M2+) as Lewis acid are reported. Experimental results and theoretical calculations indicate the peroxidase-mimicking activity of MxV2O5·nH2O show a periodic change with the Lewis acidity (ion potential) of M2+, revealing the mechanism of redox-inert M2+ regulating electron transfer of V-O through non-covalent polarization and thus promoting H2O2 adsorbate dissociation. The biomimetic synergetic effect of redox site and Lewis acid is expected to provide an inspiration for design of enzyme mimics.

Abstract DNase‐catalyzed hydrolysis of extracellular DNA (eDNA) has been widely employed to eradicate intractable biofilms. Although aggregation‐induced emission (AIE) has become the ideal tool for killing planktonic bacteria, AIE luminogens (AIEgens) often lack DNase‐mimetic activity, so as to suffer from poor anti‐biofilm capacity. Here, an “AIEzyme”, a kind of AIE nanomaterial with enzyme‐like activity, is designed and synthesized, where the AIEgens are used as the ligands of Zr‐based coordination polymer nanoparticles. Not only does AIEzyme have enduring DNase‐mimetic activity with low activation energy, but also structural rigidity‐stabilized fluorescence. Due to the long‐acting hydrolysis for eDNA in biofilm, AIEzyme can efficiently disorganize the established biofilms with good penetrability and realize the healing of superbug‐infected wounds under only one dose of AIEzyme. This work provides a strategy to endow ordinary AIEgens with DNase‐like and anti‐biofilm activities. Moreover, AIEzymes can be observed by virtue of their own AIE character, facilitating the study on self‐positioning and residual amount of AIEzymes in wounds. The concept “AIEzyme” would hopefully stimulate the tremendous expansion for the tool kits and the application of AIEgens and artificial enzymes.

Current phototherapeutic approaches for Alzheimer's disease (AD) exhibit restricted clinical outcomes due to the limited physical penetration and comprised brain microenvironment of noninvasive nanomedicine. Herein, a hydrogen-bonded organic framework (HOF) based sonosensitizer is designed and synthesized. Mn-TCPP, a planar molecule where Mn 2+ ion is chelated in the core with a large p-conjugated system and 4 carboxylate acid groups, has been successfully used as building blocks to construct an ultrasound-sensitive HOF (USI-MHOF), which can go deep in the brain of AD animal models. The both in vitro and in vivo studies indicate that USI-MHOF can generate singlet oxygen ( 1 O 2 ) and oxidize β-amyloid (Aβ) to inhibit aggregation, consequently attenuating Aβ neurotoxicity. More intriguingly, USI-MHOF exhibits catalase (CAT)- and superoxide dismutase (SOD)-like activities, mitigating neuron oxidative stress and reprograming the brain microenvironment. For better crossing the blood-brain barrier (BBB), the peptide KLVFFAED (KD8) has been covalently grafted to USI-MHOF for improving BBB permeability and Aβ selectivity. Further, in vivo experiments demonstrate a significant reduction of the craniocerebral Aβ plaques and improvement of the cognition deficits in triple-transgenic AD (3×Tg-AD) mice models following deep-penetration ultrasound treatment. The work provides the first example of an ultrasound-responsive biocompatible HOF as non-invasive nanomedicine for in-depth treatment of AD., (© 2024 Wiley‐VCH GmbH.)

Nanomaterials excel in mimicking the structure and function of natural enzymes while being far more interesting in terms of structural stability, functional versatility, recyclability, and large‐scale preparation. Herein, the story assembles hemin, histidine analogs, and G‐quadruplex DNA in a catalytically competent supramolecular assembly referred to as assembly‐activated hemin enzyme (AA‐heminzyme). The catalytic properties of AA‐heminzyme are investigated both in silico (by molecular docking and quantum chemical calculations) and in vitro (notably through a systematic comparison with its natural counterpart horseradish peroxidase, HRP). It is found that this artificial system is not only as efficient as HRP to oxidize various substrates (with a turnover number kcat of 115 s−1) but also more practically convenient (displaying better thermal stability, recoverability, and editability) and more economically viable, with a catalytic cost amounting to <10% of that of HRP. The strategic interest of AA‐heminzyme is further demonstrated for both industrial wastewater remediation and biomarker detection (notably glutathione, for which the cost is decreased by 98% as compared to commercial kits). [ABSTRACT FROM AUTHOR]

The introduction of an abiological catalytic group into the binding pocket of a protein host allows for the expansion of enzyme chemistries. Here, we report the generation of an artificial enzyme by genetic encoding of a non‐canonical amino acid that contains a secondary amine side chain. The non‐canonical amino acid and the binding pocket function synergistically to catalyze the asymmetric nitrocyclopropanation of α,β‐unsaturated aldehydes by the iminium activation mechanism. The designer enzyme was evolved to an optimal variant that catalyzes the reaction at high conversions with high diastereo‐ and enantioselectivity. This work demonstrates the application of genetic code expansion in enzyme design and expands the scope of enzyme‐catalyzed abiological reactions. [ABSTRACT FROM AUTHOR]

Abstract Oral dihydroxyphenylalanine (Dopa) administration to replenish neuronal dopamine remains the most effective treatment for Parkinson’s disease (PD). However, unlike the continuous and steady dopamine signaling in normal neurons, oral Dopa induces dramatic fluctuations in plasma Dopa levels, leading to Dopa-induced dyskinesia. Herein, we report a functional nucleic acid-based responsive artificial enzyme (FNA-Fe3O4) for in situ continuous Dopa production. FNA-Fe3O4 can cross the blood-brain barrier and target diseased neurons relying on transferrin receptor aptamer. Then, FNA-Fe3O4 responds to overexpressed α-synuclein mRNA in diseased neurons for antisense oligonucleotide treatment and fluorescence imaging, while converting to tyrosine aptamer-based artificial enzyme (Apt-Fe3O4) that mimics tyrosine hydroxylase for in situ continuous Dopa production. In vivo FNA-Fe3O4 treatment results in recovery of Dopa and dopamine levels and decrease of pathological overexpressed α-synuclein in PD mice model, thus ameliorating motor symptoms and memory deficits. The presented functional nucleic acid-based responsive artificial enzyme strategy provides a more neuron friendly approach for the diagnosis and treatment of PD.

Artificial enzymes are valuable biocatalysts able to perform new-to-nature transformations with the precision and (enantio-)selectivity of natural enzymes. Although they are highly engineered biocatalysts, they often cannot reach catalytic rates akin those of their natural counterparts, slowing down their application in real-world industrial processes. Typically, their designs only optimise the chemistry inside the active site, while overlooking the role of protein dynamics on catalysis. In this work, we show how the catalytic performance of an already engineered artificial enzyme can be further improved by distal mutations that affect the conformational equilibrium of the protein. To this end, we subjected a specialised artificial enzyme based on the lactococcal multidrug resistance regulator (LmrR) to an innovative algorithm that quickly inspects the whole protein sequence space for hotpots which affect the protein dynamics. From an initial predicted selection of 73 variants, two variants with mutations distant by more than 11 Å from the catalytic pAF residue showed increased catalytic activity towards the new-to-nature hydrazone formation reaction. Their recombination displayed a 66% higher turnover number and 14 °C higher thermostability. Microsecond time scale molecular dynamics simulations evidenced a shift in the distribution of productive enzyme conformations, which are the result of a cascade of interactions initiated by the introduced mutations.

Pneumonia involves complex immunological and pathological processes leading to pulmonary dysfunction, which can be life-threatening yet lacks effective specialized medications. Natural enzymes can be used as biological agents for the treatment of oxidative stress-related diseases, but limiting to catalytic and environmental stability as well as high cost. Herein, an artificial enzyme, gold nanoclusters (Au NCs) with excellent stability, bioactivity, and renal clearance can be used as the next-generation biological agents for acute lung injury (ALI) and allergic lung disease (ALD). The Au 25 clusters can mimic catalase (CAT) and glutathione peroxidase (GPx), and the K m of Au 24 Er 1 with H 2 O 2 reaches 1.28 mM, about 22 times higher than natural CAT (≈28.8 mM). The clusters inhibit the oxidative stress in the mitochondria and promote the synthesis of adenosine triphosphate (ATP). The molecular mechanism shows that the TLR4/MyD88/NF-κB pathway and M1 macrophage-mediated inflammatory response are suppressed in ALI and the Th1/Th2 imbalance in ovalbumin (OVA)-induced ALD is rescued. Further, the clusters can notably improve lung function in both ALI and ALD models which paves the way for immunomodulation and intervention for lung injury and can be used as a substitute for natural enzymes and potential biopharmaceuticals in the treatment of various types of pneumonia., (© 2024 Wiley‐VCH GmbH.)

We have approached the construction of an artificial enzyme by employing a robust protein scaffold, lactococcal multidrug resistance regulator, LmrR, providing a structured secondary and outer coordination spheres around a molecular rhodium complex, [RhI(PEt2NglyPEt2)2]−. Previously, we demonstrated a 2–3 fold increase in activity for one Rh-LmrR construct by introducing positive charge in the secondary coordination sphere. In this study, a series of variants was made through site-directed mutagenesis where the negative charge is located in the secondary sphere or outer coordination sphere, with additional variants made with increasingly negative charge in the outer coordination sphere while keeping a positive charge in the secondary sphere. Placing a negative charge in the secondary or outer coordination sphere demonstrates decreased activity by a factor of two compared to the wild-type Rh-LmrR. Interestingly, addition of positive charge in the secondary sphere, with the negatively charged outer coordination sphere restores activity. Vibrational and NMR spectroscopy suggest minimal changes to the electronic density at the rhodium center, regardless of inclusion of a negative or positive charge in the secondary sphere, suggesting another mechanism is impacting catalytic activity, explored in the discussion. [ABSTRACT FROM AUTHOR]

Nanoparticles that catalyze biochemically relevant reactions are promoted as alternative enzymes. The application of such artificial enzymes is severely restricted by poor selectivity in biological fluids; mainly because the reactions occur at active sites on the exterior surface of the nanoparticle. Enzymes in contrast typically have their active sites down a nanoconfined substrate channel where the reaction occurs in different solution conditions to bulk solution which aids in achieving selectivity for the substrate. Herein the same 3D structure of enzymes is mimicked in nanoparticles to allow selective reactions in biological fluids. This is achieved using a gold nanoparticle coated in a conducting mesoporous carbon shell where isolated nanochannels lead to the gold surface. It can detect glucose in whole blood with no interference from other species. This is achieved by electrochemically pulsing the artificial enzymes to generate the locally required alkalinity for an effective electrocatalytic reaction in the nanochannels, as well as expelling fouling agents that will otherwise passivate the electrocatalytic reaction. The artificial enzymes are shown to be capable of detecting glucose in biological fluids, without loss of signal, for several months. This study shows how nanoconfinement in nanoparticles can be exploited to potentially allow a broad range of species to be selectively detected in biological fluids with stability that can exceed that of enzymes. [ABSTRACT FROM AUTHOR]

Nitrogenase catalyzes substrate reduction at its cofactor center ([(Cit)MoFe7S9C](n-); designated M-cluster). Here, we report the formation of an artificial, nitrogenase-mimicking enzyme upon insertion of a synthetic model complex ([Fe6S9(SEt)2](4-); designated Fe6(RHH)) into the catalytic component of nitrogenase (designated NifDK(apo)). Two Fe6(RHH) clusters were inserted into NifDK(apo), rendering the conformation of the resultant protein (designated NifDK(Fe)) similar to the one upon insertion of native M-clusters. NifDK(Fe) can work together with the reductase component of nitrogenase to reduce C2H2 in an ATP-dependent reaction. It can also act as an enzyme on its own in the presence of Eu(II)DTPA, displaying a strong activity in C2H2 reduction while demonstrating an ability to reduce CN(-) to C1-C3 hydrocarbons in an ATP-independent manner. The successful outcome of this work provides the proof of concept and underlying principles for continued search of novel enzymatic activities based on this approach.

The active site accessibility and high loading of gold nanoparticles (AuNPs) are key factors affecting the catalytic activity of supported AuNP-based catalysts. However, the preparation of supported AuNP-based catalysts with highly accessible active sites still remains a challenge. Herein, sphere-on-sphere (SoS) silica microspheres with a hierarchical structure, good dispersion and high surface density of thiol groups (10 SH nm -2 ) are prepared and used as a platform for the growth of high-density AuNPs. The obtained hierarchical Si@Au micro-/nanostructure consisting of 0.55 μm SoS silica microspheres and 7.3 nm AuNPs (SoS-0.55@Au-7.3) is found to show excellent peroxidase-mimicking activity ( K m = 0.033 mM and V max = 34.6 × 10 -8 M s -1 ) with merits of high stability and good reusability. Furthermore, the as-obtained SoS-0.55@Au-7.3-based system can sensitively detect hydrogen peroxide (H 2 O 2 ) with a low detection limit of 1.6 μM and a wide linear range from 2.5 μM to 1.0 mM. The high catalytic activity, excellent stability and good reusability of SoS-0.55@Au-7.3 imply its great prospects in biosensing and biomedical analysis., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

The rational design of catalysts with enzyme-like properties is an elusive goal of chemists despite tremendous interest. Molecular imprinting inside surfactant micelles, followed by postmodification, creates a tailored active site in a water-soluble polymeric "artificial enzyme" for the benzylation of 4-nitrophenol. The reaction happens under neutral conditions with excellent substrate selectivity. Similar to many enzymes, electrostatics play vital roles in catalysis and can be tuned through different bases introduced into the active site.

With the rapid evolution of sensing technologies, the integration of nanoscale catalysts, particularly those mimicking enzymatic functions, into electrochemical devices has surfaced as a pivotal advancement. These catalysts, dubbed artificial enzymes, embody a blend of heightened sensitivity, selectivity, and durability, laying the groundwork for innovative applications in real-time health monitoring and environmental detection. This minireview penetrates into the fundamental principles of electrochemical sensing, elucidating the unique attributes that establish artificial enzymes as foundational elements in this field. We spotlight a range of innovations where these catalysts have been proficiently incorporated into wearable and portable platforms. Navigating the pathway of amalgamating these nanoscale wonders into consumer-appealing devices presents a multitude of challenges; nevertheless, the progress made thus far signals a promising trajectory. As the intersection of materials science, biochemistry, and electronics progressively intensifies, a flourishing future seems imminent for artificial enzyme-infused electrochemical devices, with the potential to redefine the landscapes of wearable health diagnostics and portable sensing solutions.

Bacteria therapy, using large sizes bacteria equipped with flagella, could easily penetrate the tumor tissue, exerting amazing effects. Inspired by this, carbon nanotubes‐assisted strategy, using CNTs to simulate the flagella of bacteria, is developed to improve the ability of vector to enter the cell. Via bionics, tumor microenvironment (TME) specific‐activated composite photothermal/Fenton enzyme (PZCo‐800) reacted selectively on the tumor sites was developed via direct pyrolysis of a Co‐Porphyrin‐based conjugated porous polymer (CMP) coated zeolitic‐imidazolate‐framework (ZIF‐8) hybrid. Carbonization caused the formation of flagella‐like CNTs, improving the ability of particles entering the cancer cells. PZCo‐800 utilized the acidic and hydrogen peroxide (H2O2) to initiate the Fenton‐like reaction (CDT), producing reactive oxygen species (ROS) to kill tumor cells. Additionally, under laser irradiation (638 nm), the heat generated by PZCo‐800 offered a prominent photothermal therapy (PTT) boosting the Fenton‐like therapeutic effect. Both in vitro and in vivo assays verified the PTT and CDT worked collaboratively with a remarkable inhabitation of 81.17 % on tumor growth, far outperforming the monomodal CDT and PTT. This microenvironment activated artificial enzyme enabled safe and reliable cancer combination therapy. [ABSTRACT FROM AUTHOR]