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Lithium iron phosphate materials as one of the most representative cathode materials for lithium-ion battery were found to possess intrinsic peroxidase-like activity. LiFePO4 particles were synthesized by a classic hydrothermal process. In this paper, colorless 3,3',5,5'-tetramethylbenzidine is oxidized to a blue product using hydrothermally synthesized LiFePO4 particles (LFP) as a catalyst with a significant absorption peak at 650 nm. Kinetic assay showed that the catalytic performance was consistent with typical Michaelis–Menten kinetics. As a novel peroxidase mimic material, LFP show the advantages of high catalytic efficiency and favorable stability in aqueous solution compared with natural enzymes, thus providing a simple, inexpensive and fast approach to colorimetric detection of H2O2 and glucose.

Enantioselective protonation is conceptually one of the most attractive methods to generate an α-chiral center. However, enantioselective protonation presents major challenges, especially in water as a solvent. Herein, we report an artificial enzyme catalyzed tandem Michael addition and enantioselective protonation reaction of α-substituted acroleins with 2-acyl imidazole derivatives in water. The artificial enzyme uses a synergistic combination of two abiological catalytic sites: a genetically encoded non-canonical p-aminophenylalanine residue and a Lewis acid Cu(II) complex. The exquisite stereochemical control achieved in the protonation of the transient enamine intermediate generated by conjugate addition of the Michael donor is illustrated by the >20:1 dr and up to >99% ee obtained for the products. These results illustrate the potential of exploiting synergistic catalysis in artificial enzymes for challenging reactions.

Based on a biomimetic point of view, a new artificial enzyme model was designed to imitate the biocatalytic behavior of native-chloroperoxidase hemoenzyme. Transition metals catalyzed C–H chlorination and represented the C–H activation process. Cysteine–metal complexes of Mn(II), Fe(III), Co(II), Ni(II), Cu(II) and Zn(II) were prepared and encapsulated in a sodium dodecyl sulfate (SDS)/dodecyl trimethyl ammonium bromide (DTAB) vesicular mixture to imitate a native-chloroperoxidase model at pH 3. Current biocatalysts also behaved as a catalase at pH 3 and as a peroxidase at pH 1. Surprisingly, “Cu(II)–cysteine/SDS/DTAB” nanozyme indicated the maximum chloroperoxidase, catalase and peroxidase activity or efficacy among other mentioned intermediate metal complexes including manganese (2+), cobalt (2+), zinc (2+), iron (3+) and nickel (2+). The performed structural analysis by transmission electron microscopy and dynamic light scattering illustrated different nanozyme shapes and surface charges at distinct pHs. Zeta potential of colloids indicated the presence of Cu(II)–cysteine active site inside the nanozyme. Differential scanning calorimetry shows different stabilities and fluidities of nanozyme at different pHs. FTIR spectroscopy was utilized to investigate the structural complexation of Cu(II)–cysteine active essence. The present study demonstrated that “the sulfur” moiety of l-cysteine is ligated to copper ion at pH 1 and 3 in order to form Cu(II)–cysteine center as a multi-functional active site. Accordingly, the current study presents the preparation of an efficient pseudo-vesicular Cu(II)–cysteine supramolecular system with pH-switchable multi-functional enzymatic ability.

The interplay between nucleic acids and lipids underpins several key processes in molecular biology, synthetic biotechnology, vaccine technology, and nanomedicine. These interactions are often electrostatic in nature, and much of their rich phenomenology remains unexplored in view of the chemical diversity of lipids, the heterogeneity of their phases, and the broad range of relevant solvent conditions. Here we unravel the electrostatic interactions between zwitterionic lipid membranes and DNA nanostructures in the presence of physiologically relevant cations, with the purpose of identifying new routes to program DNA-lipid complexation and membrane-active nanodevices. We demonstrate that this interplay is influenced by both the phase of the lipid membranes and the valency of the ions and observe divalent cation bridging between nucleic acids and gel-phase bilayers. Furthermore, even in the presence of hydrophobic modifications on the DNA, we find that cations are still required to enable DNA adhesion to liquid-phase membranes. We show that the latter mechanism can be exploited to control the degree of attachment of cholesterol-modified DNA nanostructures by modifying their overall hydrophobicity and charge. Besides their biological relevance, the interaction mechanisms we explored hold great practical potential in the design of biomimetic nanodevices, as we show by constructing an ion-regulated DNA-based synthetic enzyme.

Herein, a robust Pd(II)-based polyfunctional magnetic amphiphilic artificial metalloenzyme was prepared by anchoring a Pd(2,2′-dipyridylamine)Cl2 bearing hydrophilic monomethyl ether poly(ethylene glycol) (mPEG) chains on the surface of amino-functionalized silica-coated magnetic nanoparticles. The 2,2′-dipyridylamine (dpa) has shown excellent complexation properties for Pd(II) and it could be easily anchored onto functionalized magnetic support by the bridging nitrogen atom. Moreover, the bridging nitrogen atom at the proximity of Pd(II) catalytic center could play an important role in dynamic suppramolecular interactions with substrates. The leaching, air and moisture resistant [Pd(dpa)Cl2] complex endow the dynamic and robust structure to the designed artificial enzyme. Moreover, the water dispersibility of designed artificial metalloenzyme raised from mPEG chains and the magnetic nanoparticles core which could function as protein mimics endow it other necessary characters of artificial enzymes. The prepared artificial metalloenzyme displayed remarkable activity in Suzuki–Miyaura cross-coupling reaction employing low-palladium loading under mild conditions, with the exceptionally high turnover frequency, clean reaction profile, easy work-up procedure, good to excellent products yields and short reaction times. The designed air- and moisture-stable artificial metalloenzyme could recycle more than fifteen times with easy separation procedure in aqueous solution under aerobic conditions without any noticeable loss in activity.

Site-isolation of catalysts can enable incompatible catalysts such as acid and base to be used in one pot for enhanced efficiency and other benefits. Although many synthetic platforms have been reported for this purpose, they generally do not possess the exquisite selectivity of site-isolated enzymes in nature. Here we report water-soluble protein-sized nanoparticles with site-isolated acids in the core and amines on the surface. The catalysts were made through molecular imprinting of cross-linked micelles, followed by facile one-step photoaffinity labeling of the imprinted binding site. With a tunable, substrate-specific active site, the bifunctional artificial enzyme catalyzed highly selective tandem cross aldol reaction between acetone and mixtures of isomeric aryl acetals. It could also transform a less reactive substrate over a more reactive one.

Natural enzymes are efficient and versatile biocatalysts but suffer in their environmental tolerance and catalytic stability. As artificial enzymes, nanozymes can improve the catalytic stability, but it is still a challenge to achieve high catalytic activity. Here, we employed atomic engineering to build the artificial enzyme named Au24Ag1 clusterzyme that hosts an ultrahigh catalytic activity as well as strong physiological stability via atom manipulation. The designed Au24Ag1 clusterzyme activates the Ag-S active site via lattice expansion in the oligomer atom layer, showing an antioxidant property 72 times higher than that of natural antioxidant Trolox. Enzyme-mimicked studies find that Au24Ag1 clusterzyme exhibits high catalase-like (CAT-like) and glutathione peroxidase-like (GPx-like) activity with a maximum reaction rate of 68.9 and 17.8 μM/min, respectively. Meanwhile, the unique catalytic landscape exhibits distinctive reactions against inflammation by inhibiting the cytokines at an early stage in the brain. Atomic engineering of clusterzymes provides a powerful and attractive platform with satisfactory atomic dispersion for tailoring biocatalysts freely at the atomic level.

Synthetic dyes such as malachite green (MG) have a wide range of applications. Meanwhile, they bring great challenges for environmental security and cause potential damages to human health. Compared with traditional approaches, enzymatic catalysis is an emerging technique for wastewater treatment. As alternatives to natural enzymes, artificial enzymes have received much attention for potential applications. In previous studies, we have rationally designed artificial enzymes based on myoglobin (Mb), such as by introducing a distal histidine (F43H mutation) and creating a channel to the heme pocket (H64A mutation). We herein show that the artificial enzyme of F43H/H64A Mb can be successfully applied for efficient biodegradation of MG under weak acid conditions. The degradation efficiency is much higher than those of natural enzymes, such as dye-decolorizing peroxidase and laccase (13–18-fold). The interaction of MG and F43H/H64A Mb was investigated by using both experimental and molecular docking studies, and the biodegradation products of MG were also revealed by UPLC-ESI-MS analysis. Based on these results, we proposed a plausible biodegradation mechanism of MG. With the high-yield of overexpression in E. coli cells, this study suggests that the artificial enzyme has potential applications in the biodegradation of MG in fisheries and textile industries.

A Metal-Organic Framework (MOFs) with large surface area, exposed active site, excellent catalytic performance and high chemical stability has been used as an artificial enzyme and designed for nonenzymatic electrochemical sensors. Here, a strategy of using an enhanced electrochemical sensing platform for the detection of nitic oxide (NO) and hydrogen peroxide (H2O2) was designed via a nano-metalloporphyrinic metal-organic framework (NporMOF(Fe)) as an electrode material. By taking advantage of the small size, high surface area and exposed Fe active site, the obtained NporMOF(Fe) displays excellent electrocatalytic activity toward NO and H2O2. The NporMOF(Fe) modified electrode shows high sensing ability toward the in situ generated NO in NO2- containing phosphate buffer (PB) solution with a wide linear detection range of 5 μM to 200 μM and a very low detection limit of 1.3 μM. Moreover, NporMOF(Fe) exhibits high electrocatalytic activity toward the reduction of H2O2 and the practical detection of H2O2 released from HeLa cells. Furthermore, the NporMOF(Fe) modified electrode shows excellent selectivity toward the detection of NO and H2O2 in the presence of other physiologically important analytes. This method shows excellent biosensing performance, implying the universal applicability of MOFs-based artificial nanozymes for biosensors and the potential application for third generation biosensors.

Peroxidase widely exists in nature and can be applied for the diagnosis and detection of H2O2, glucose, ascorbic acid and other aspects. However, the natural peroxidase has low stability and its catalytic efficiency is easily affected by external conditions. In this work, a copper-based metal–organic framework (Cu-MOF) was prepared by hydrothermal method, and characterized by means of XRD, SEM, FT-IR and EDS. The synthesized Cu-MOF material showed high peroxidase-like activity and could be utilized to catalyze the oxidation of o-phenylenediamine (OPDA) and 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H2O2. The steady-state kinetics experiments of the oxidation of OPDA and TMB catalyzed by Cu-MOF were performed, and the kinetic parameters were obtained by linear least-squares fitting to Lineweaver–Burk plot. The results indicated that the affinity of Cu-MOF towards TMB and OPDA was close to that of the natural horseradish peroxidase (HRP). The as-prepared Cu-MOF can be applied for colorimetric detection of H2O2 and glucose with wide linear ranges of 5 to 300 μM and 50 to 500 μM for H2O2 and glucose, respectively. Furthermore, the specificity of detection of glucose was compared with other sugar species interference such as sucrose, lactose and maltose. In addition, the detection of ascorbic acid and sodium thiosulfate was also performed upon the inhibition of TMB oxidation. Based on the high catalytic activity, affinity and wide linear range, the as-prepared Cu-MOF may be used for artificial enzyme mimics in the fields of catalysis, biosensors, medicines and food industry.

Realizing a sustainable future requires intensifying the waste stream conversion, such as converting the greenhouse gas carbon dioxide into value-added products. In this paper, we focus on utilizing formaldehyde as a C1 carbon source for enzymatic C–C bond formation. Formaldehyde can be sustainably derived from other C1 feedstocks, and in this work, we designed a synthetic enzyme cascade for producing the functional C4 sugar erythrulose. This involved tailoring the enzyme formolase, which was optimized for fusing formaldehyde, from a three-carbon producer (dihydroxyacetone) to sets of variants with enhanced two-carbon (glycolaldehyde) or four-carbon (erythrulose) activity. To achieve this, a high-throughput combinatorial screening was developed, and every single variant was evaluated in terms of glycolaldehyde, dihydroxyacetone and erythrulose activity. By applying the two most promising variants in an enzyme cascade, we were able to show for the first time production of ERY starting from a C1 carbon source. In addition, we demonstrated that one of our tailored formolase variants was able to convert 25.0 g L−1 glycolaldehyde to 24.6 g L−1 erythrulose (98% theoretical yield) in a fully atom-economic biocatalytic process. This represents the highest achieved in vitro concentration of erythrulose to date.

Background Earlier studies have attempted to create electronic free insulin delivery systems using different glucose sensing mechanism, no successful clinical translation as hitherto been made. This study aimed to assess the faster responsiveness of the insulin release from this enzyme based nanoparticles which is a self-regulated insulin delivery system constructed by loading with insulin, enzyme glucose oxidase into hyaluronic acid and 2-nitroimidazole forming enzyme-based nanoparticles which works in accordance to the blood glucose level. Materials and method Enzyme-based nanoparticles were prepared by ionic gelation method. Insulin content in the nanoparticles kept for stability study was estimated by human insulin enzyme based immunosorbent assay. In in-vitro studies; different concentrations of glucose were taken and the release study of insulin was recorded. Results This enzyme-based nanoparticles were having average diameter of around 193 nm and stability studies showed that nanoparticles were stable upto 30 days at 4 °C. In-vitro studies showed the release of insulin from nanoparticle conjugates which was effectively correlated with the external glucose concentration created where different concentrations of glucose taken thus facilitating the stabilization of blood glucose levels in the hyperglycemia state which was achieved within 10 min. (400 mg/dL) wherein drug release rate remarkably increased in hyperglycemia state and no specific changes or small amount of release was observed in normoglycemia state (100 mg/dL). Conclusion Overall, this preliminary study of this enzyme-based nanoparticles formulation showed excellent rapid responsiveness towards hyperglycemia which might act as a potential biomimetic system in triggering the release of insulin in sustained manner.

International audience; Performing a heterogeneous catalysis with proteins is still a challenge. Herein, we demonstrate the importance of cross‐linked crystals for sulfoxide oxidation by an artificial enzyme. The biohybrid consists of the insertion of an iron complex into a NikA protein crystal. The heterogeneous catalysts displays a better efficiency‐with higher reaction kinetics, a better stability and expand the substrate scope compared to its solution counterpart. Designing crystalline artificial enzymes represents a good alternative to soluble or supported enzymes for the future of synthetic biology.

Nanozymes as one of artificial enzymes show many advantages than natural enzymes. The high Michaelis-Menten constant (Km) to H2O2 is the drawback for nanozymes, which means a high H2O2 concentration to oxidize 3,3',5,5'-tetramethylbenzidine (TMB). For this problem, FeS2/SiO2 double mesoporous hollow spheres (DMHSs) were first synthesized as an artificial peroxidase through a solid reaction. The experimental results demonstrate that Fe3O4 vulcanization and DMHS formation were effective strategies to enhance affinity to H2O2 for the nanozyme. The Km of FeS2/SiO2 DMHSs (H2O2 as the substrate) is 18-fold smaller than that of FeS2 nanoparticles (NPs). The catalytic efficiency (Kcat/Km) of FeS2/SiO2 DMHSs is about 16 times higher than that of FeS2 NPs. FeS2/SiO2 DMHSs can be used as a nanozyme to sensitively and rapidly detect H2O2 and glutathione within 1 min at room temperature.

Developing artificial enzyme mimetics for the detection of small biomolecules are a current research interest because natural enzymes bear some serious disadvantages, such as their catalytic activity can be easily inhibited and they can be digested by proteases. Herein, a heteropoly acids (HPA) encapsulating metal–organic framework (MOF) with metal-carbene structure, [Cu10(H3trz)4(Htrz)4][PMo12VO41] (PMA-MOF) as bifunctional enzyme-mimetic catalyst for colorimetric detection of hydrogen peroxide (H2O2) and ascorbic acid (AA) was designed and synthesized. Thanks to the good stability and the synergistic effect of PMA and MOF, PMA-MOF exhibits the lower limit of detection (0.222 μM towards H2O2 and 0.0046 μM to AA), and the smaller Km value (0.0138 mM for H2O2 and 0.136 mM for o-phenylenediamine) compared to most reported MOF- and HPA-based enzyme-mimetic catalyst, to the best our knowledge.

Artificial enzyme mimics are more stable and cost-effective than bioenzymes, but they usually lack substrate recognition function. Here we report that uranyl (UO22+) features efficient artificial p...

Polymeric carbon nitride (CN) is one of the most promising metal-free photocatalysts to alleviate the energy crisis and environmental pollution. Loading cocatalysts is regarded as an effective way to improve the photocatalytic efficiency of CNs. However, commonly used noble metal cocatalysts limit their applications due to their rarity and high cost. Herein, we present the effective synthesis of single-atom copper-modified CN via supramolecular preorganization with subsequent condensation, which provides effective charge transfer pathways by an "infused" delocalized state with variable-valence catalysis at the same time. The C-Cu-N2 single-atom catalytic site can activate CO2 molecules and reduces the energy barrier toward photocatalytic CO2 reduction. Excellent performance for photocatalytic CO2 reduction was found. This work thereby provides a general protocol of designing a noble-metal-free photocatalyst with infused metal centers toward a wide range of applications.

Acetylcholinesterase inactivating compounds, such as organophosphate (OP) and carbamate (CM) pesticides, are widely used in agriculture to ensure sustainable production of food and feed. As a consequence of their applications, they would result in neurotoxicity, even death. In this essence, the development of enzyme inhibition methods still shows great significance as rapid detection techniques for on-site large-scale screening of OPs and CMs. Initially, mechanisms and applications of various enzyme-inhibition-based methods and devices, including optical colorimetric assay, fluorometric assays, electrochemical biosensors, rapid test card, and microfluidic device, are highlighted in the present overview. Further, to enhance the enzyme sensitivity for detection; alternative enzyme sources or high yield enrichment methods (such as abzyme, artificial enzyme, and recombinant enzyme), as well as enzyme reactivation and identification, are also addressed in this comprehensive overview.

Dinuclear Cu-2(II,II)-cores stabilized by the N3O donorset of HL1 = (2-{[[di(2-pyridyl)methyl](methyl)amino]methyl}phenol), HL2 = 2-({[di(2-pyridyl)methyl] amino}methyl)phenol), and HL3 = 2-({[di(2-pyridyl)methyl]amino}methyl)-4-nitrophenol display a unique superoxide dismutase (SOD) combined with catalase (CAT)-like activity in water, at neutral pH. The Cu2L21 < Cu2L22 < Cu2L23 structure-reactivity trend puts a spotlight on the electron-deficient core of Cu2L23 that exhibits the highest SOD (log k(cat) (O-2(center dot-)) = 7.55) and CAT-like (k(H2O2) = 0.66 M(-1)s(-1)) performance. Time-lapse ESI-MS and EPR experiments indicate that a dimeric core is essential for oxygenic turnover upon H2O2 decomposition.

A high degree of dependence on fossil fuels is one of the major problems faced by modern societies. d-Glucose and glycerol have emerged in recent years as prospective replacements for fossil fuels ...

Staphylococcus aureus (S. aureus) is an important foodborne pathogen that causes food poisoning and severe infections. Herein, we design a novel and sensitive colorimetric immunoassay for the detection of S. aureus based on the formation of a magnetic bead-based sandwich complex and signal amplification via an enzyme mimic. Magnetic nanoparticles decorated with carbon dots offer rapid enrichment and separation of target bacteria in complex matrices. Sliver nanoclusters are used as an artificial enzyme that can oxidize o-phenylenediamine to form a yellow product with a maximum absorption peak at 421 nm. This method can visually detect S. aureus ranging from 10 to 106 cfu mL−1. Its limit of detection reaches down to 4.9 cfu mL−1. By analyzing food samples, the recovery is from 85.6 to 103.7%.

A systematic structure–activity relationship study was performed to identify the factors that are important to enhancing the transfer hydrogenation efficiency of half-sandwich metal complexes, which is an important class of compounds recently shown to be useful as artificial enzyme cofactors, catalytic drugs, and intracellular catalysts. In general, electron-rich ligands provided catalysts with greater activity and chemical stability than electron-poor ligands under physiologically relevant aqueous conditions. Complexes containing Ir were better catalysts than those containing Ru, Rh, or Os. Kinetic studies by UV-vis absorption spectroscopy revealed that ligand substituent effects could dramatically accelerate the rates of both hydride formation and hydride transfer processes, the latter by up to a remarkable 28-fold. The thermodynamic hydricities of several iridium–hydride species were determined using a hydride exchange method, which showed a strong correlation between the hydride donor strengths of the iridium–hydride species and the transfer hydrogenation yields of their parent complexes. Specifically, this work provides a practical guide for further elaboration of the pentamethylcyclopentadienyl iridium pyridinecarboxamidate catalyst platform. We found that electron-donating groups on the pyridyl ring are conducive to achieving high catalytic rates whereas a wide variety of alkyl and aryl groups on the N-amide moiety are well-tolerated, suggesting that this position might be the best site for conjugation to other functional entities for biological applications.

The color generating from the biochemical reaction between 3,3′,5,5′-tetramethylbenzidine and Lysine@WS2 QDs was used a signal for the detection of hydrogen peroxide. The QDs were prepared using a combination of techniques, that is, probe sonication and hydrothermal treatment. Analysis via UV–vis spectroscopy, Fourier transform infrared and Raman spectroscopy, X-ray diffraction, energy-dispersive spectroscopy, and transmission electron microscopy yielded detailed information on the nature and characteristics of these quantum dots. Furthermore, as-synthesized quantum dots were studied for their capability to mimic peroxidase enzyme using 3,3′,5,5′-tetramethylbenzidine as a substrate. Consequently, a colorimetric sensor utilizing Lysine@WS2 QDs could detect hydrogen peroxide in a range of 0.1–60 μM with a response time of 5 min. The same material was used for H2O2 detection using impedance spectroscopy, which yielded a dynamic range of 0.1–350 μM with a response time of 30–40 s.

A new hybrid nanoparticle (NP; fluorenylmethoxycarbonyl-arginine-glycine-aspartate and hemin, Fmoc-RGD/hemin NP) was developed for the simultaneous detection and inhibition of breast cancer cells. Hemin can regulate the reactive oxygen species (ROS), while Fmoc-RGD acts as a scaffold for hemin nanocrystallization. Fmoc groups interact with the porphyrin groups of hemin through hydrophobic and π-π interactions to form a hydrophobic core of the NPs. The hydrophilic RGD chains surround the core to maintain the stability of the nanoparticles in an aqueous medium. The RGD groups of Fmoc-RGD are also selective for tumor cells. This interaction can be exploited to enhance the selectivity of tumor detection. Based on enhanced peroxidase activity, Fmoc-RGD/hemin NPs were developed as signal transducers in a facile and fast point-of-care cancer diagnosis platform. This platform is sensitive to breast cancer cells and hydrogen peroxide (H

L-Histidine (L-His) is an essential amino acid, which is used to synthesize proteins and enzymes. The concentration of L-His in the body is controlled to regulate tissue growth and repair of tissues. In this study, a rapid and sensitive method was developed for colorimetric L-his detection using Cu2+ ions to inhibit the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB)–H2O2 system. H2O2 can oxidize TMB to oxTMB in the presence of copper, and the change in color from colorless (TMB) to blue (oxTMB) is similar to that observed in the presence of peroxidase. However, because the imidazole ring and carboxyl group of L-His can coordinate with Cu2+ ions to form stable L-His–Cu2+ complexes, the color of the TMB–H2O2 solution remains unchanged after the addition of L-His. Therefore, because L-His effectively hinders the colorimetric reaction of TMB with H2O2, this assay can be used to quantitatively determine the concentration of L-His in samples. Under optimized conditions, our colorimetric sensor exhibited two linear ranges of 60 nM to 1 μM and 1 μM to 1 mM for L-His detection and a detection limit of 50 nM (S/N = 3); furthermore, the assay can be performed within 20 min. Moreover, the proposed assay was used to determine the concentration of L-His in urine samples, suggesting that this convenient and label-free colorimetric method presents promising applications in bioanalytical chemistry and clinical diagnosis.

Tumor-related exosomes, which are heterogeneous membrane-enclosed nanovesicles shed from cancer cells, have been widely recognized as potential noninvasive biomarkers for early cancer diagnosis. Herein, an artificial enzyme cascade amplification strategy based on a switchable DNA tetrahedral (SDT) scaffold was proposed for quantification of breast cancer-derived exosomes. The SDT scaffold is composed of G-quadruplex mimicking DNAzyme sequences on its two single-stranded edges and glucose oxidase (GOx) on the four termini of the complementary strands. In the initial state, the SDT scaffold is blocked by the switch strand which consists of partial complementary domains with the DNA tetrahedron and a MUC1 aptamer. MCF-7 exosomes could release the quadruplex-forming sequences through the recognition of the MUC1 aptamer. The newly formed DNAzyme brings GOx into spatial proximity and induces high-efficiency enzyme cascade catalytic reactions on the SDT. Consequently, high sensitivity toward MCF-7 exosome analysis was obtained with a wide linear range of 3.8 × 106 to 1.2 × 108 particles per mL and a limit of detection of 1.51 × 105 particles per mL. In addition, such a DNAzyme reconfiguration strategy was able to distinguish MCF-7 exosomes from other breast cancer cell derived exosomes, indicating its excellent method specificity. The proposed enzyme cascade strategy not only provides a novel signal transformation and amplification nanoplatform for quantifying the specific populations of exosomes, but also can be further expanded to the analysis of multiple cancer biomarkers.

Aspartic proteases use a pair of carboxylic acids to activate water molecules for nucleophilic attack. Here we report a nanoparticle catalyst with a similar catalytic motif capable of generating a hydroxide ion in its active site even under acidic reaction conditions. The synthetic enzyme accelerated the hydrolysis of para-nitrophenyl acetate (PNPA) by 91,000 times and could also hydrolyze nonactivated aryl esters at pH 7. The distance between the two acids and, in particular, the flexibility of the catalytic groups in the active site controlled the catalytic efficiency. The synthetic enzyme readily detected the addition of a single methyl on the acyl group of the substrate, as well as the substitution pattern on the phenyl ring.

Cell-free extract and purified enzyme-based systems provide an attractive solution to study biosynthetic strategies towards a range of chemicals. 4-(4-hydroxyphenyl)-butan-2-one, also known as raspberry ketone, is the major fragrance component of raspberry fruit and is used as a natural additive in the food and sports industry. Current industrial processing of the natural form of raspberry ketone involves chemical extraction from a yield of ∼1–4 mg kg−1 of fruit. Due to toxicity, microbial production provides only low yields of up to 5–100 mg L−1. Herein, we report an efficient cell-free strategy to probe into a synthetic enzyme pathway that converts either L-tyrosine or the precursor, 4-(4-hydroxyphenyl)-buten-2-one, into raspberry ketone at up to 100% conversion. As part of this strategy, it is essential to recycle inexpensive cofactors. Specifically, the final enzyme step in the pathway is catalyzed by raspberry ketone/zingerone synthase (RZS1), an NADPH-dependent double bond reductase. To relax cofactor specificity towards NADH, the preferred cofactor for cell-free biosynthesis, we identify a variant (G191D) with strong activity with NADH. We implement the RZS1 G191D variant within a ‘one-pot’ cell-free reaction to produce raspberry ketone at high-yield (61 mg L−1), which provides an alternative route to traditional microbial production. In conclusion, our cell-free strategy complements the growing interest in engineering synthetic enzyme cascades towards industrially relevant value-added chemicals.

In this work, a simple one-step thermal oxidation process was established to achieve a significant surface increase in {110} and {111} nanofacets on well-defined, pure and Pr-doped, ceria nanocubes. More importantly, without changing most of the bulk properties, this treatment leads to a remarkable boost of their enzymatic activities: from the oxidant (oxidase-like) to antioxidant (hydroxyl radical scavenging) as well as the paraoxon degradation (phosphatase-like) activities. Such performance improvement might be due to the thermally generated sawtoothlike {111} nanofacets and defects, which facilitate the oxygen mobility and the formation of oxygen vacancies on the surface. Finally, possible mechanisms of nanoceria as artificial enzymes have been proposed in this manuscript. Considering the potential application of ceria as artificial enzymes, this thermal treatment may enable the future design of highly efficient nanozymes without changing the bulk composition., This work has been supported by the Ministry of Science, Innovation and Universities of Spain with Reference Numbers of ENE2017-82451-C3-2-R, MAT2016-81118-P and MAT2017-87579-R. The research projects funded by the Natural Science Foundation of Shandong Province (Grant ZR2017LB028), Key R&D Program of Shandong Province (Grant 2018GSF118032), and Fundamental Research Funds for the Central Universities (Grant 18CX02125A) in China are also acknowledged. TEM/STEM data were obtained at DMEUCA node of the Spanish Unique Scientific and Technological Infrastructure (ICTS) of Electron Microscopy of Materials ELECMIM. M. Tinoco thanks the FPU Scholarship Program (Grant AP2010-3737) from Ministry of Education of Spain. H. Pan is grateful for financial support (Grant 201406140130) from the Chinese Scholarship Council to accomplish her Ph.D. study at the University of Cadiz (Spain). J. M. Gonzalez, G. Blanco, and X. Chen are also grateful for the financial support from the joint project (Proyectos Integradores, Grant PI20201) in IMEYMAT of the University of Cadiz.

Highly-efficient and eco-friendly materials and technologies are urgently needed to meet the requirements of nowadays green development. Photocatalysis with using solar energy and enzymatic catalysis with eco-friendly nature are effective alternatives to address the problem. Notably, beneficial use of the synergistic effect of artificial enzyme and advanced photocatalyst has attracted wide attention. This work presents a biomimetic photocatalytic material, two-dimensional (2D) biomimetic hemin-bismuth tungstate (HBWO). Stable HBWO composites formed by immobilization of monomeric hemin on 2D bismuth tungstate layer, exhibit high photocatalytic performance, better than that of pure 2D bismuth tungstate and unsupported hemin. HBWO shows layered structure with the interlayer spacing at ˜0.35 nm. In the photocatalytic process, hemin can not only act as an electron shuttle, also play an important role in oxygen transfer. Additionally, the synthesized HBWO composites exhibit nice binding affinities and high photocatalytic activity in tetracycline degradation. It is anticipated that beneficial use of synergistic effect of artificial enzyme and photocatalyst via HBWO composites can be a promising eco-friendly and efficient solution for addressing the environmental crisis.

Streptavidin (SA), and other related proteins, has been isolated from a wide range of organisms, including bacteria, fungi, frogs, fish, and birds. Although their original function is not well understood, they have found an important place in biotechnology based on their unique ability to bind biotin molecules with high affinity and specificity. The SA-biotin interaction is robust and easy to incorporate into different designs, and as such, it is used when reliable molecule interaction is needed under poorly controlled experimental conditions. There are continued efforts to engineer these proteins to modulate their size, valency, and affinity, since the optimum molecular properties vary depending on individual applications. This review will describe recent developments in streptavidin engineering to meet these requirements, including those that form novel oligomeric states, e.g., a monomer, have fewer functional biotin-binding sites, or bind biotin with reduced affinity. We also examine various reported applications of both natural or engineered SA constructs in cell biology, biochemistry, genetics, synthetic chemistry, cancer therapy, drug delivery, and nanotechnology to illustrate the breadth of modern science that is advanced by the endogenous and engineered SA-biotin interactions.

Glucaric acid (GlucA) has been identified as one of the top 10 potential bio-based chemicals for replacement of oil-based chemicals. Several synthetic enzyme pathways have been engineered in bacteria and yeast to produce GlucA from glucose and myo-inositol. However, the yields and titres achieved with these systems remain too low for the requirements of a bio-based GlucA industry. A major limitation for the optimisation of GlucA production via synthetic enzymatic pathways are the laborious analytical procedures required to detect the final product (GlucA) and pathway intermediates. We have developed a novel method for the simple and simultaneous analysis of GlucA and pathway intermediates to address this limitation using mixed mode (MM) HILIC and weak anion exchange chromatography (WAX), referred to as MM HILIC/WAX, coupled with RID. Isocratic mobile phase conditions and the sample solvent were optimised for the separation of GlucA, glucose-1-phosphate (G1P), glucose-6-phosphate (G6P), inositol-1-phosphate (I1P), myo-inositol and glucuronic acid (GA). The method showed good repeatability, precision and excellent accuracy with detection and quantitation limits (LOD and LOQ) of 1.5–2 and 577 mM, respectively. The method developed was used for monitoring the enzymatic synthesis of the final step in the GlucA pathway, and showed that GlucA was produced from GA with near 100% conversion and a titre of 9.2 g L−1.

Due to oil depletion and global climate change, sustainable manufacturing of fine chemicals from renewable feedstocks has gained increasing attention in the scientific community. In the present stu...

Outcomes of chemical reactions are generally dominated by the intrinsic reactivities of reaction partners, but enzymes frequently override such constraints to transform less reactive molecules in the presence of more reactive ones. Despite the attractiveness of such catalysis, it is difficult to build synthetic catalysts with these features. Micellar imprinting is a powerful method to create template-complementary binding sites inside protein-sized water-soluble nanoparticles. When a photocleavable functional monomer was used to bind two phosphonate/phosphate templates as transition-state analogues, active sites with predetermined size and shape were formed inside doubly cross-linked micelles through molecular imprinting. Postmodification replaced the binding group with a catalytic pyridyl group, forming highly selective artificial esterases. The catalysts displayed enzyme-like kinetics and turnover numbers that were in the hundreds. The selectivity of the catalysts, derived from the substrate-complementary imprinted active sites, enabled transformation of less reactive esters in the presence of more reactive ones.

To develop a highly sensitive colorimetric detection method for the diabetes early diagnosis by determining the glucose in urine instead of blood can avoid the painful blood collection and risks of cross infection. In this paper, novel artificial enzyme mimics were formed by loading bimetallic nanoparticle Fe Pd on the surface of reduced graphene (rGO). Also, we proposed a Fe Pd/rGO coupled with portable paper sensor for the detection of glucose in urine. The results indicated that urine glucose can be sensitively detected in a wide range of 1–200 μM with a limit of detection (LOD) of 1.76 μM. Most important, the color difference along with the increase of urine glucose concentration can be easily distinguished by naked eyes, which is important to its practical usage in screening and diagnosis of diabetes. Furthermore, the reason of the high sensitivity of Fe Pd/rGO towards glucose was investigated in detail via various probing experiments. Fe Pd/rGO has an artificial enzyme mimic activity, which can catalyze O2 and generate H2O2 from glucose oxidation. The electron transfer between Fe Pd and H2O2 will produce •OH that can oxidize terephthalic acid (TMB) by emitting a blue color. The findings in this study provide a promising alternative for the early screening and diagnosis of diabetes.

The construction of a novel bioanode based on l-proline oxidation using a cascade reaction pathway comprised of thermostable dehydrogenases. A novel multi-enzymatic cascade pathway, containing four kinds of dehydrogenases from thermophiles (dye-linked l-proline dehydrogenase, nicotinamide adenine dinucleotide (NAD)-dependent Δ1-pyrroline-5-carboxylate dehydrogenase, NAD-dependent l-glutamate dehydrogenase and dye-linked NADH dehydrogenase), was designed for the generation of six-electrons from one molecule of l-proline. The current density of the four-dehydrogenase-immobilized electrode, with a voltage of + 450 mV (relative to that of Ag/AgCl), was 226.8 μA/cm2 in the presence of 10 mM l-proline and 0.5 mM ferrocene carboxylate at 50 °C. This value was 4.2-fold higher than that of a similar electrode containing a single dehydrogenase. In addition, about 54% of the initial current in the multi-enzyme cascade bioanode was maintained even after 15 days. Efficient deep oxidation of l-proline by multiple-enzyme cascade reactions was achieved in our designed electrode. The multi-enzyme cascade bioanode, which was built using thermophilic dehydrogenases, showed high durability at room temperature. The long-term stability of the bioanode indicates that it shows great potential for applications as a long-lived enzymatic fuel cell.

We report herein, a novel two dimensional (2D) nanocomposite sensor towards detection of glucose via a simple colorimetric detection technique. The graphitic carbon nitride (g-C3N4) sheets are decorated with Au-Ni bimetallic nanoparticles with size ˜11 nm by a simple chemical reduction technique. The Au-Ni/g-C3N4 nanocomposites is found to possess intrinsic peroxidase like activity capable of catalysing the oxidation of 3,3ʹ,5,5ʹ-tetramethylbenzidine (TMB) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) in presence of H2O2 and exhibits substrate affinity much higher than biological horseradish peroxidase as well as other reported nanozymes. Also, there is a scanty of literature reports on bimetallic g-C3N4 based nanozymes towards detection of any biomolecules related to human life. The peroxidase like activity combined with glucose oxidase (GOx) helped in developing a simple, sensitive and a specific colorimetric method for glucose detection with a detection limit of 1.7 μM and a linear detection range between 0.5‒30 μM. The colorimetric method for detection of glucose was further successfully extended towards detection of glucose in real serum samples with comparable results to standard hospital method. Additionally, the nanocomposites are non-cytotoxic towards human umbilical vein endothelial cell line (HUVEC) which demonstrates its high biocompatibility and suitability for biosensor applications. Our results thus demonstrate that our method based on Au-Ni/g-C3N4 is reliable, economical, feasible and highly efficient.

Metal–organic frameworks with diverse structures and unique properties have demonstrated that can be an ideal substitute for natural enzymes in colorimetric sensing platform for analyte detection in various fields such as environmental chemistry, biotechnology and clinical diagnostics, which have attracted the scientist’s attention, recently. In this study, a porous coordination network (denoted as PCN-222) was synthesized as a new biomimetic material from an iron linked tetrakis (4-carboxyphenyl) porphyrin (named as Fe-TCPP) as a heme-like ligand and Zr6 linker as a node. This catalyst shows the peroxidase and catalase activities clearly. The mechanism of peroxidase activity for PCN-222 was investigated using the spectrophotometric methods and its activity was compared with the other nanoparticles which, the results showed a higher activity than the other catalysts. Also, the hydrogen peroxide was detected by PCN-222(Fe) based on the peroxidase-like activity. For detection of hydrogen peroxide a linear range of 3–200 μM and detection of limit (LOD) 1 μM (3σ/slope), under optimal conditions were obtained. Moreover, based on the high tendency of PCN-222(Fe) to combine with the TMB as a chromogenic substrate in the peroxidase-like activity, we developed the sensitive and selective colorimetric assay for glucose detection that was found a detection limit (LOD) of 2.2 μM in the linear range from 12 to 75 μM. Finally due to the good catalytic activity of PCN-222(Fe), it was used to detection of glucose and hydrogen peroxide in real samples.

Nanostructured artificial enzyme mimics (nanozymes) are promising frontiers and have received considerable attention in chemical research. However, the practical applications of nanozymes were severely hampered by their limited catalytic activity and overall catalytic efficiency, and the current reports indicated that quite rare vanadium oxides have oxidase activity. Herein, we demonstrated that fluoride capped V6O13–rGO nanocomposites exhibited high oxidase mimetic activity for the first time. The well fluoride capped V6O13–rGO nanocomposites obviously improved the catalytic activity and kinetic performance compared with V6O13 and V6O13–rGO. The Vmax of V6O13–rGO in the presence of fluoride for TMB oxidation was 10−8 M s−1, which was 2.8-fold higher than that of V6O13–rGO without fluoride, illustrating a better efficiency substrate uptake. The mechanism is attributed to the specific fluoride capping, which could change the surface charge of V6O13–rGO for improving the substrate adsorption affinity and enhancing the electron transfer rate. The prepared nanozyme exhibited excellent catalytic performance for the establishment of fluoride colorimetric sensors and detection of fluoride in tap water. The outcome indicated that the developed nanozyme could be extended to a broad range of practical applications in biotechnology, biocatalysis, and biomedical engineering.

Herein, a rigid 3D DNA nanopillar was used to investigate the influence of spatial organization on the cascade activity in multienzyme systems, realizing controllable regulation of the mimic enzyme ratio and spacing for acquiring a high-efficiency enzyme cascade catalytic platform. Initially, the ratio of mimic enzyme AuNPs (glucose oxidase-like activity) and hemin/G-quadruplex DNAzyme (peroxidase-like activity) fixed at the designed position was adjusted by changing the number of edges in a DNA polyhedron, resulting in an optimal mimic enzyme ratio of 1 : 4 with a quadrangular prism as the scaffold. Notably, the DNA nanopillar formed by quadrangular prism layer-by-layer assembly acted as a track for directional and controllable movement of a bipedal DNA walker based on the toehold mediated strand displacement reaction (TSDR), which endowed the assay system with continuous enzyme spacing regulation compared with previous enzyme cascade systems that induced inflexible operation. Furthermore, enzyme mimetics in this work circumvented the drawbacks of natural enzymes, such as time-consuming purification processes and poor thermal stability. As a proof of concept, the proposed dual regulation strategy of cascade enzymes was applied in the ultrasensitive electrochemical detection of Pb2+, which provided a new route to fabrication of high-performance artificial enzyme cascade platforms for ultimate application in bioanalysis and biodiagnostics., A rigid 3D DNA nanopillar was used to investigate the influence of spatial organization on the cascade activity in multienzyme systems, realizing controllable regulation of the mimic enzyme ratio and spacing for efficient cascade catalytic platform.

Heme is both an essential cofactor and an abundant source of nutritional iron for the human pathogen Mycobacterium tuberculosis (Mtb). While heme is required for Mtb survival and virulence, it is also potentially cytotoxic. Since Mtb has the ability to both make and uptake heme, the de novo synthesis of heme and its acquisition from the host must be balanced in order to mitigate heme toxicity. However, the mechanisms employed by Mtb to regulate heme uptake, synthesis, and bioavailability are poorly understood. By integrating ratiometric heme sensors with mycobacterial genetics, cell biology, and biochemistry, we determined that the terminal heme biosynthetic enzyme, coproheme decarboxylase (ChdC), plays a role in regulating both heme bioavailability and uptake in Mtb. Moreover, we found that Mtb has a preference for scavenging reduced ferrous heme and exhibits a cell surface heme reductase activity that is regulated by ChdC. In Mtb, ChdC expression is down-regulated when iron is limiting, which in-turn increases both heme import and bioavailability. Such a mechanism may serve to protect cells from heme toxicity while trying to meet the nutritional demand for iron. Our results demonstrate that heme synthesis and uptake are tightly integrated in mycobacteria and represent the first example of a heme synthetic enzyme playing a role in controlling heme uptake. Significance Statement Heme is an essential but potentially cytotoxic cofactor and iron source for the pathogen, Mycobacterium tuberculosis (Mtb). To understand how Mtb coordinates heme uptake and synthesis to mitigate heme toxicity, we integrated heme sensors with mycobacterial genetics and biochemical approaches to probe the interplay between heme synthesis and scavenging. We discovered that the terminal heme synthetic enzyme, coproheme decarboxylase (ChdC), negatively regulates heme uptake and utilization in response to iron availability through a mechanism involving control of a ferric heme reductase. During iron limitation, ChdC is downregulated, thereby enhancing exogenous heme reduction, uptake and utilization while simultaneously suppressing heme synthesis, which allows Mtb to avoid heme toxicity. Our results highlight the close coordination between heme synthesis and uptake in mycobacteria. Classification Biological sciences : Biochemistry

The conversion of Kraft lignin in plant biomass into renewable chemicals, aiming at harvesting aromatic compounds, is a challenge process in biorefinery. Comparing to the traditional chemical methods, enzymatic catalysis provides a gentle way for the degradation of lignin. Alternative to natural enzymes, artificial enzymes have been received much attention for potential applications. We herein achieved the biodegradation of Kraft lignin using an artificial peroxidase rationally designed in myoglobin (Mb), F43Y/T67R Mb, with a covalently linked heme cofactor. The artificial enzyme of F43Y/T67R Mb has improved catalytic efficiencies at mild acidic pH for phenolic and aromatic amine substrates, including Kraft lignin and the model lignin dimer guaiacylglycerol-β-guaiacyl ether (GGE). We proposed a possible catalytic mechanism for the biotransformation of lignin catalyzed by the enzyme, based on the results of kinetic UV-Vis studies and UPLC-ESI-MS analysis, as well as molecular modeling studies. With the advantages of F43Y/T67R Mb, such as the high-yield by overexpression in E. coli cells and the enhanced protein stability, this study suggests that the artificial enzyme has potential applications in the biodegradation of lignin to provide sustainable bioresource.

Catalytic cascade transformations, which occur in spatially constrained tumor environment to generate therapeutic moieties from prodrugs or intrinsic species, are highly desirable for precise cancer therapy. Nevertheless, it is high challenging to engineer a cascade nanoreactor with tumor microenvironment (TME)-responsive capability for synergistic tumor therapy. Inspired by the biocatalytic cascades in biological processes, here, a tumor-specific nanoreactor was established to activate cascade reactions for oxidative stress-augmented chemotherapy by the integration of an artificial enzyme, Pt(IV)-based prodrug (Pt(IV)), with Cu(II)-based metal-organic frameworks (CuMOF). Upon internalization of CuMOF@Pt(IV) by tumor cells, in addition to chemotherapeutic effect, the activated cisplatin by glutathione (GSH) reduction is capable of acting as an artificial enzyme to elevate the hydrogen peroxide (H2O2) level through cascade reactions for augmenting the therapeutic efficacy of Cu+-mediated chemodynamic therapy (CDT). Meanwhile, CuMOF@Pt(IV) specifically deplete overexpressed GSH at tumor sites, thus amplifying tumor oxidative stress, and finally leading to augmented antitumor efficacy. The orchestrated cooperative effect of chemotherapy and oxidative stress presents splendid therapeutic efficacy on tumor-bearing mice with negligible adverse effects. Therefore, this cascade nanoreactor provides exciting opportunities to develop complementary therapeutic modalities for precise cancer treatment.

To evaluate the antioxidant activity of potential synthetic enzyme mimetics, we prepared new five copper(II) complexes via a self-assembly method and named them [Cu(2-(HOCH2)py)3](ClO4)2 (1), [Cu(2-(HOCH2)py)2(H2O)2]SiF6 (2), [Cu2(2-(HOCH2CH2)py)2(2-(OCH2CH2)py)2](ClO4)2 (3), [Cu(pyBIm)3](BF4)2·1.5H2O (4) and [Cu(py2C(OH)2)2](ClO4)2 (5). The synthetic protocol involved N,O- or N,N-donors: 2-(hydroxymethyl)pyridine (2-(HOCH2)py), 2-(hydroxyethyl)pyridine (2-(HOCH2CH2)py), 2-(2-pyridyl)benzimidazole (pyBIm), di(2-pyridyl)ketone (py2CO). The obtained Cu(II) complexes were fully characterised by elemental analysis, FTIR, EPR, UV-Vis, single-crystal X-ray diffraction and Hirshfeld surface analysis. Crystallographic and spectroscopic analyses confirmed chromophores of both monomeric ({CuN3O3} (1), {CuN2O4} (2), {CuN6} (4), {CuN4O2} (5)) and dimeric complex ({CuN2O3} (3)). Most of the obtained species possessed a distorted octahedral environment, except dimer 3, which consisted of two copper centres with square pyramidal geometries. The water-soluble compounds (1, 3 and 5) were selected for biological testing. The results of the study revealed that complex 1 in solutions displayed better radical scavenging activity than complexes 3, 5 and free ligands. Therefore, complex 1 has been selected for further studies to test its activity as an enzyme mimetic. The chosen compound was tested on the erythrocyte lysate of two groups of patients after undergoing chemotherapy and chemoradiotherapy. The effect of the tested compound (1) on enzyme activity levels (TAS, SOD and CAT) suggests that the selected complex can be treated as a functional mimetic of the enzymes.

Engineering of a promiscuous lactonase via semi-rational evolution gave a 1007-fold improvement in its catalytic activity in the degradation of triphenyl phosphate (TPHP). TPHP is a typical bulky organophosphate flame retardant (OPFR) and is widely used in industry. To the best of our knowledge, this is the first artificial enzyme capable of degrading OPFRs.

A novel ferrocene-linked metal-covalent organic polymer (MCOP-NFC) was synthesized through the Claisen–Schmidt condensation reaction of 1,1′-diacetyl ferrocene and tris(4-formylphenyl)amine. MCOP-NFC acts as a highly efficient artificial enzyme for mimicking peroxidase, and shows good stability in harsh chemical environments including strong bases and acids, and boiling water. Based on the peroxidase-like activity of MCOP-NFC, a highly sensitive dual channel detection method for hydrogen peroxide was developed. For the colorimetric detection strategy, the limit of detection (LOD) reached 2.1 μM, while the limit of detection was found to be as low as 0.08 μM based on the electrochemical detection channel. This study offers a new strategy for the development of an enzyme mimetic on the basis of the covalent assembly of nanostructures, and the proposed electrochemical–colorimetric sensor for H2O2 detection has great potential for applications in biology and biomedicine.

We report the facile and economic preparation of two-dimensional (2D) and 0D MoSe2 nanostructures based on systematic and non-toxic top-down strategies. We demonstrate the intrinsic peroxidase-like activity of these MoSe2 nanostructures. The catalytic processes begin with facilitated decomposition of H2O2 by using MoSe2 nanostructures as peroxidase mimetics. In turn, a large amount of generated radicals oxidizes 3,3,5,5-tetramethylbenzidine (TMB) to produce a visible color reaction. The enzymatic kinetics of our MoSe2 nanostructures complies with typical Michaelis&ndash, Menten theory. Catalytic kinetics study reveals a ping&ndash, pong mechanism. Moreover, the primary radical responsible for the oxidation of TMB was identified to be Ȯ2&minus, by active species-trapping experiments. Based on the peroxidase mimicking property, we developed a new colorimetric method for H2O2 detection by using 2D and 0D MoSe2 nanostructures. It is shown that the colorimetric sensing capability of our MoSe2 catalysts is comparable to other 2D materials-based colorimetric platforms. For instance, the linear range of H2O2 detection is between 10 and 250 &mu, M by using 2D functionalized MoSe2 nanosheets as an artificial enzyme. Our work develops a systematic approach to use 2D materials to construct novel enzyme-free mimetic for a visual assay of H2O2, which has promising prospects in medical diagnosis and food security monitoring.

Metalloenzymes such as the carbonic anhydrases (CAs, EC 4.2.1.1) possess highly specialized active sites that promote fast reaction rates and high substrate selectivity for the physiologic reaction that they catalyze, hydration of CO2 to bicarbonate and a proton. Among the eight genetic CA macrofamilies, α-CAs possess rather spacious active sites and show catalytic promiscuity, being esterases with many types of esters, but also acting on diverse small molecules such as cyanamide, carbonyl sulfide (COS), CS2, etc. Although artificial CAs have been developed with the intent to efficiently catalyse non-biologically related chemical transformations with high control of stereoselectivity, the activities of these enzymes were much lower when compared to natural CAs. Here, we report an overview on the catalytic activities of α-CAs as well as of enzymes which were mutated or artificially designed by incorporation of transition metal ions. In particular, the distinct catalytic mechanisms of the reductase, oxidase and metatheses-ase such as de novo designed CAs are discussed.

In this report, the streptavidin-biotin technology was applied to enable organocatalytic aldol addition. By attaching pyrrolidine to the valeric motif of biotin and introducing it to streptavidin (Sav), a protein-based organocatalytic system was created, and the aldol addition of acetone with p-nitrobenzaldehyde was tested. The conversion of substrate to product can be as high as 93%. Although the observed enantioselectivity was only moderate (33:67 er), further protein engineering efforts can be included to improve the selectivity. These results have proven the concept that Sav can be used to host stereoselective aldol addition.

Artificial enzyme mimics are of current research interest owing to their remarkable advantages over natural enzymes. Herein, as a novel peroxidase mimic material, MIL-101(Cr)@PB was fabricated by encapsulating Prussian blue (PB) nanoparticles into the host matrix of MIL-101(Cr) via a facile and mild in situ growth synthetic strategy. The crystallographic characteristics, morphology, and porosity of the as-synthesized MIL-101(Cr)@PB composites were carefully studied using XRD, SEM, TEM, TGA, and BET. The results show that the synthesized MIL-101(Cr)@PB possesses a reproducible and impressive intrinsic peroxidase-like activity even under extreme conditions. Exploiting this, a colorimetric platform for screening xanthine oxidase inhibitors was constructed. We hope that this work will elucidate the applications of metal–organic frameworks as carriers for enzyme mimics and enable a wider application in drug screening.