Nanozymes (NZs) are nanostructured artificial enzymes that mimic catalytic properties of natural enzymes. The NZs have essential advantages over natural enzymes, namely low preparation costs, stability, high surface area, self-assembling capability, size and composition-dependent activities, broad possibility for modification, and biocompatibility. NZs have wide potential practical applications as catalysts in biosensorics, fuel-cell technology, environmental biotechnology, and medicine. Most known NZs are mimetics of oxidoreductases or hydrolases. The present work aimed to obtain effective artificial peroxidase (PO)-like NZs (nanoPOs), to characterize them, and to estimate the prospects of their analytical application. NanoPOs were synthesized using a number of nanoparticles (NPs) of transition and noble metals and were screened for their catalytic activity in solution and on electrodes. The most effective nanoPOs were chosen as NZs and characterized by their catalytic activity. Kinetic parameters, size, and structure of the best nanoPOs (Cu/CeS) were determined. Cu/CeS-based sensor for H2O2 determination showed high sensitivity (1890 A·M−1·m−2) and broad linear range (1.5–20,000 µM). The possibility to apply Cu/CeS-NZ as a selective layer in an amperometric sensor for hydrogen-peroxide analysis of commercial disinfectant samples was demonstrated.
Metal nanoparticles (NPs), such as gold (Au) and silver (Ag), are important for chemistry, physics, and biology due to their unique optical, electrical, and photothermal properties. Such NPs are widely used for immobilization of various bioactive substances, including peptides, enzymes, antibodies and DNA. The synthesis of silver and gold nanoparticles was carried out by reduction of silver nitrate by glucose and reduction of tetrachloroauric acid by sodium citrate, respectively. The size and structure of the AgNPs and AuNPs were characterized using TEM, AFM and XRD methods. The average size of the AgNPs and AuNPs was between 8 and 15 nm. Recombinant arginase I was immobilized using the carbodiimidepentafluorophenol method on the surface of NPs functionalized with ω-mercaptohexadecanoic acid. It was shown that recombinant human liver arginase I isolated from the yeast Hansenula polymorpha maintains satisfactory stability after immobilization on both NPs. The immobilized arginase retained 40% of its activity on the surface of AuNPs and 25% on AgNPs compared to the free arginase after storage at +4 ºC during 25 days. The immobilized enzyme can be used for assay of arginine in pharmaceuticals, in food products and in blood
Novel nanomaterials, including metallic nanoparticles obtained via green synthesis (gNPs), have a great potential for application in biotechnology, industry and medicine. The special role of gNPs is related to antibacterial agents, fluorescent markers and carriers for drug delivery. However, application of gNPs for construction of amperometric biosensors (ABSs) is not well documented. The aim of the current research was to study potential advantages of using gNPs in biosensorics. The extracellular metabolites of the yeast Ogataea polymorpha were used as reducing agents for obtaining gNPs from the corresponding inorganic ions. Several gNPs were synthesized, characterized and tested as enzyme carriers on the surface of graphite electrodes (GEs). The most effective were Pd-based gNPs (gPdNPs), and these were studied further and applied for construction of laccase- and alcohol oxidase (AO)-based ABSs. AO/GE, AO-gPdNPs/GE, laccase/GE and laccase-gPdNPs/GE were obtained, and their analytical characteristics were studied. Both gPdNPs-modified ABSs were found to have broader linear ranges and higher storage stabilities than control electrodes, although they are less sensitive toward corresponding substrates. We thus conclude that gPdNPs may be promising for construction of ABSs for enzymes with very high affinities to their substrates.
Prussian blue analogs (PBAs) are well-known artificial enzymes with peroxidase (PO)-like activity. PBAs have a high potential for applications in scientific investigations, industry, ecology and medicine. Being stable and both catalytically and electrochemically active, PBAs are promising in the construction of biosensors and biofuel cells. The “green” synthesis of PO-like PBAs using oxido-reductase flavocytochrome b2 is described in this study. When immobilized on graphite electrodes (GEs), the obtained green-synthesized PBAs or hexacyanoferrates (gHCFs) of transition and noble metals produced amperometric signals in response to H2O2. HCFs of copper, iron, palladium and other metals were synthesized and characterized by structure, size, catalytic properties and electro-mediator activities. The gCuHCF, as the most effective PO mimetic with a flower-like micro/nano superstructure, was used as an H2O2-sensitive platform for the development of a glucose oxidase (GO)-based biosensor. The GO/gCuHCF/GE biosensor exhibited high sensitivity (710 A M−1m−2), a broad linear range and good selectivity when tested on real samples of fruit juices. We propose that the gCuHCF and other gHCFs synthesized via enzymes may be used as artificial POs in amperometric oxidase-based (bio)sensors.
The binary phases Ti 5 M 3 , Ti 3 M and Zr 3 M (M = Sn, Sb) were studied for electrochemical lithiation, using powder X-ray diffraction, scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDX). The investigation showed that the morphology of the cathode and the anode surfaces undergo changes, and the grain size of the materials decreases. The phase analysis of the anode materials revealed that the Ti 5 Sn 3 (structure type Mn 5 Si 3 ) and Ti 3 Sn (structure type Mg 3 Cd) phases form solid solutions by insertion of Li atoms into the initial structure. The insertion is reversible. The phases Ti 5 Sb 3 (structure type Y 5 Bi 3 ), Ti 3 Sb, Zr 3 Sn (structure type Cr 3 Si), and Zr 3 Sb (structure type Ni 3 P) form solid solutions by substitution of Li for Sn or Sb atoms. Only the Zr 3 Sb phase showed weakly reversible substitution. Among the investigated compounds, the most suitable structure types for intercalation of lithium appeared to be the Mn 5 Si 3 -and Mg 3 Cd-types, where the Li atoms occupy octahedral voids. The intermetallic compounds containing tin showed better ability for electrochemical lithiation than the compounds containing antimony. This can be explained by the easier interaction of antimony and lithium with the formation of binary compounds.
Intermetallic compound / Electrochemical lithiation / Li-ion battery
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