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Physicochemical and enzymatic methods of quantitative L-arginine estimation are described. A variety of detection procedures for L-arginine analysis have been developed. The majority of the frequently used approaches are marked by poor precision, low sensitivity and selectivity. These methods are time-consuming, expensive and require skilful labor techniques, so it needs to develop novel highly selective and sensitive ones for improvement of L-arginine monitoring in clinical diagnostics and food industry. Experimental data concerning development and testing of effective enzymatic methods of L-arginine determination, including biosensors, are presented. All proposed methods are based on human liver arginase I isolated from the recombinant yeast strain Hansenula polymorpha NCYC-495 pGAP1-HsARG1 (leu2car1 Sc:LEU2). Arginase I (EC 3.5.3.1; L-arginine amidinohydrolase), a key enzyme of the urea cycle, catalyses the conversion of L-arginine to ornithine and urea. An effectiveness of the proposed enzymatic methods for L-arginine assay, as amperometric and potentiometric biosensors so as enzymatic methods with spectrophotometric and fluorometric detection of the product, was demonstrated on the samples of commercial pharmaceuticals. These methods seem to be prospective for L-arginine analyses in food industry (juices and wines) and in medicine, for diagnostics and drug control in blood serum under treatment of cancer malignances. Hence, the proposed L-Arg selective and simple methods would be convenient and useful in the future for routine clinical analysis and food quality control.

This work aims to develop new H2O2-sensing elements and nanozyme-oxidase-based sensors for the detection of hydrogen peroxide, glucose, and galactose. Methods. Chemical synthesis. Assay of enzymatic activity. Scanning electron microscope. Results. Hexacyanoferrates of several transient metals were synthesized and tested for their nanozymes (HCF NZs) (certificial peroxidase) activity in solution. The best representative nanozymes (NZs) were used for the construction of H2O2-sensitive sensors and nanozyme-oxidase-based biosensors. The NZs-based sensors: nAuHCF/GE and nPtHCF/GE demonstrated higher sensitivity (7.5 and 9.4-fold) than the biosensor with natural peroxidase. The developed biosensors GOX/nPtHCF/GE and GaOx/nPtHCF/GE for the detection of glucose and galactose showed enhanced sensitivity: 900 and 540 A·M-1·m-2, respectively, broad linear range (0.02-0.2 mM). Along with a broad linear range of detection these biosensors possess low limits of detection -- 4.0 μM for glucose and 6.0 μM for galactose. Additionally, these biosensors exhibited improved stability when compared to the control. Conclusions. The novelty of the presented work is related to the synthesis of new peroxidase-like NZs and the evaluation of their functionality as the chemosensors on H2O2 and as the sensing components in the oxidase-based-biosensors for assay of glucose and galactose. The nPtHCF and nAuHCF NZs exhibit high sensitivity for the target analytes, a broad linear range, and satisfactory storage stability. The main advantages of the proposed nanozyme-oxidase-based biosensors are the simple architecture of the sensing layer and the ability to operate at low working electrode potentials. [ABSTRACT FROM AUTHOR]

The predicted dynamics of soil cover degradation are verified by a matrix model, system processing of multitemporal large scaled soil maps, and satellite images. The validity of the criterion for the dynamics of typical solonchak areas to assess the soil degradation intensity is shown. [ABSTRACT FROM AUTHOR]

Enzymatic-spectrophotometric method for L-arginine assay based on human arginase I isolated from the recombinant yeast strain Hansenula polymorpha was developed. The new method is selective and rather simple. Linear detection range was estimated to be from 0.007 to 0.1 m? in reaction mixture, and detection limit -- 0.005 mM L-arginine. The method was tested on the real samples of L-arginine-containing commercial pharmaceuticals products. It was shown, that the estimated L-arginine contents were in a good correlation with that declared by producers, with results of referent chemical (R=0.998) and some other methods. [ABSTRACT FROM AUTHOR]

At the current stage of development of the branch of motor transport in Ukraine, satisfactory equipment of enterprises with the fixed assets, particularly motor transport enterprises, is one of the main problems. Fleet of road transport vehicles of those enterprises needs almost complete renovation, and productive-technical base for maintenance inspection and repair of transport requires complete technical re-equipment. The aim of the work is to develop theoretical approaches and practical recommendations concerning maintenance of road transport vehicles of motor transport enterprises, choice of the type of transportation means and determination of the necessary number of vehicles depending on the production plan of their operation.

Monitoring of the levels of 5-hydroxyindole-3-acetic acid (5-HIAA) is of significant importance for diagnostics of carcinoid tumors. We propose simple catalytic electrochemical sensors for the determination of 5-HIAA in urine using laccase and its mimetics. Laccase-like nanozymes (LacNZs) were synthesized via a chemical reduction, and resulting PtMn and MnO 2 nanoflowers (NFs) demonstrated laccase-like activity similar to the laccase from the Trametes zonata. In addition, these LacNZs showed enhanced stability under a wide range of pH (3.0-7.5), temperatures (4-70 °C), and ionic strengths (up to 500 mM NaCl). The developed PtMn NF/graphite electrode, similar to a laccase/graphite electrode, can detect 5-HIAA with a high sensitivity (25 000 ± 12 A·M -1 ·m -2 and 1900 ± 9 A·M -1 ·m -2 , respectively) and have linear ranges of 0.3 - 15 μM and 2 - 50 μM. The sensors work at low working potentials with a detection limit of 0.16 and 1.4 μM, covering the normal and pathologic ranges of 5-HIAA (1 - 50 μM) content in urine. They have been successfully applied to 5-HIAA assay in urine samples of people with various diseases and revealed good recovery values and reproducibility. Additionally, the LacNZ-sensor has the best stability and can be used up to 20 days., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

We report the development of amperometric biosensors (ABSs) employing flavocytochrome b 2 (Fc b 2 ) coupled with nanoparticles (NPs) of noble metals on graphite electrode (GE) surfaces. Each NPs/GE configuration was evaluated for its ability to decompose hydrogen peroxide (H 2 O 2 ), mimicking peroxidase (PO) activity. The most effective nanoPO (nPO) was selected for developing ABSs targeting L-lactate. Consequently, several Fc b 2 /nPO-based ABSs with enhanced sensitivity to L-lactate were developed, demonstrating mediated ET between Fc b 2 and the GE surface. The positive effect of noble metal NPs on Fc b 2 -based sensor sensitivity may be explained by the synergy between their dual roles as both PO mimetics and electron transfer mediators. Furthermore, our findings provide preliminary data that may prompt a re-evaluation of the mechanism of L-lactate oxidation in Fc b 2 -mediated catalysis. Previously, it was believed that L-lactate oxidation via Fc b 2 catalysis did not produce H 2 O 2 , unlike catalysis via L-lactate oxidase. Our initial research revealed that the inclusion of nPO in Fc b 2 -based ABSs significantly increased their sensitivity. Employing other PO mimetics in ABSs for L-lactate yielded similar results, reinforcing our hypothesis that trace amounts of H 2 O 2 may be generated as a transient intermediate in this reaction. The presence of nPO enhances the L-lactate oxidation rate through H 2 O 2 utilization, leading to signal amplification and heightened bioelectrode sensitivity. The proposed ABSs have been successfully tested on blood serum and fermented food samples, showing their promise for L-lactate monitoring in medicine and the food industry.

Introducing heterozygous humanized tau to App NL-F/NL-F knock-in mice results in the first mouse model of Alzheimer's disease in which age and amyloid-β pathology interact to initiate neurofibrillary tau tangle pathology, not dependent on mutations in MAPT. Gradual progression from amyloid-β to tau pathology in NLFTau m/h mice opens possibilities for understanding processes precipitating clinical stages of Alzheimer's disease and development of translatable therapies to prevent the onset of tau pathology.

The current research presents novel sensors based on laccase-like mimetics for the detection of dopamine (DA). The synthesized laccase-like nanozymes (nAuCu, nPtCu, nCuMnCo, and nCoCuCe) were prepared by a simple hydrothermal method and exhibited an attractive catalytic activity toward DA. The developed amperometric sensors based on laccase nanozymes (nAuCu and nPtCu) are more stable, selective, and revealed a higher sensitivity (6.5-fold than the biosensor based on the natural fungal laccase from Trametes zonata ). The amperometric sensors were obtained by modification of the glassy carbon electrodes (GCEs) with AuPt nanoparticles. Functionalization of the electrode surface by AuPt NPs resulted in increased catalytic activity of the laccase-like layer and higher sensitivity. Among studied configurations, the sensor containing nAuCu and nAuPt possesses a wide linear range for dopamine detection (10-170 μM), the lowest limit of detection (20 nM), and the highest sensitivity (10 650 ± 8.3 A M -1 m -2 ) at a low applied potential (+0.2 V versus Ag/AgCl). The proposed simple and cost-effective sensor electrode was used for the determination of DA in pharmaceuticals., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Amino acid L-arginine (Arg), usually presented in food products and biological liquids, can serve both as a useful indicator of food quality and an important biomarker in medicine. The biosensors based on Arg-selective enzymes are the most promising devices for Arg assay. In this research, three types of amperometric biosensors have been fabricated. They exploit arginine oxidase (ArgO), recombinant arginase I (ARG)/urease, and arginine deiminase (ADI) coupled with the ammonium-chelating redox-active nanoparticles. Cadmium-copper nanoparticles (nCdCu) as the most effective nanochelators were used for the development of ammonium chemosensors and enzyme-coupled Arg biosensors. The fabricated enzyme/nCdCu-containing bioelectrodes show wide linear ranges (up to 200 µM), satisfactory storage stabilities (14 days), and high sensitivities (A⋅M -1 ⋅m -2 ) to Arg: 1650, 1700, and 4500 for ADI-, ArgO- and ARG/urease-based sensors, respectively. All biosensors have been exploited to estimate Arg content in commercial juices. The obtained data correlate well with the values obtained by the reference method. A hypothetic scheme for mechanism of action of ammonium nanochelators in electron transfer reaction on the arginine-sensing electrodes has been proposed., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Catalytically active nanomaterials, in particular, nanozymes, are promising candidates for applications in biosensors due to their excellent catalytic activity, stability and cost-effective preparation. Nanozymes with peroxidase-like activities are prospective candidates for applications in biosensors. The purpose of the current work is to develop cholesterol oxidase-based amperometric bionanosensors using novel nanocomposites as peroxidase (HRP) mimetics. To select the most electroactive chemosensor on hydrogen peroxide, a wide range of nanomaterials were synthesized and characterized using cyclic voltammetry (CV) and chronoamperometry. Pt NPs were deposited on the surface of a glassy carbon electrode (GCE) in order to improve the conductivity and sensitivity of the nanocomposites. The most HRP-like active bi-metallic CuFe nanoparticles (nCuFe) were placed on a previously nano-platinized electrode, followed by conjugation of cholesterol oxidase (ChOx) in a cross-linking film formed by cysteamine and glutaraldehyde. The constructed nanostructured bioelectrode ChOx/nCuFe/nPt/GCE was characterized by CV and chronoamperometry in the presence of cholesterol. The bionanosensor (ChOx/nCuFe/nPt/GCE) shows a high sensitivity (3960 A·M -1 ·m -2 ) for cholesterol, a wide linear range (2-50 µM) and good storage stability at a low working potential (-0.25 V vs. Ag/AgCl/3 M KCl). The constructed bionanosensor was tested on a real serum sample. A detailed comparative analysis of the bioanalytical characteristics of the developed cholesterol bionanosensor and the known analogs is presented.

L-Lactate is an indicator of food quality, so its monitoring is essential. Enzymes of L-Lactate metabolism are promising tools for this aim. We describe here some highly sensitive biosensors for L-Lactate determination which were developed using flavocytochrome b 2 (Fc b 2 ) as a bio-recognition element, and electroactive nanoparticles (NPs) for enzyme immobilization. The enzyme was isolated from cells of the thermotolerant yeast Ogataea polymorpha. The possibility of direct electron transfer from the reduced form of Fc b 2 to graphite electrodes has been confirmed, and the amplification of the electrochemical communication between the immobilized Fc b 2 and the electrode surface was demonstrated to be achieved using redox nanomediators, both bound and freely diffusing. The fabricated biosensors exhibited high sensitivity (up to 1436 A·M -1 ·m -2 ), fast responses, and low limits of detection. One of the most effective biosensors, which contained co-immobilized Fc b 2 and the hexacyanoferrate of gold, having a sensitivity of 253 A·M -1 ·m -2 without freely diffusing redox mediators, was used for L-Lactate analysis in samples of yogurts. A high correlation was observed between the values of analyte content determined using the biosensor and referenced enzymatic-chemical photometric methods. The developed biosensors based on Fc b 2 -mediated electroactive nanoparticles can be promising for applications in laboratories of food control.

Nanomaterials possessing artificial, enzyme-like catalytic activity (nanozymes, NZs) have a great potential for application in research, immunological assays, biosensors, in vivo imaging, and as therapeutic agents. Despite the obvious advances in construction and understanding of functional properties of NZs, there is still no clear evidence of whether they can complement the loss of corresponding enzymatic activity in vivo. Herein, we report the first, to the best to our knowledge, example of successful substitution of natural enzyme activity by catalase-like platinum (nPt) and platinum-gold (nPtAu) nanoparticles transferred to the cells of methylotrophic yeast Ogataea polymorpha. The nPt NZs were synthesized by the chemical reduction method and used as a seed to produce the nPt(core)Au(shell) particles. The produced nPt NZs were 68.1 and 91.3 nm in size, while the hydrids were of 531.2 and 615.1 nm. Both nPt and nPtAu demonstrated catalase activity in vitro. The catalase-deficient strain Ogataea polymorpha C-105 was shown to be able to grow on methanol and a mixture of glucose and methanol in the presence although not in the absence of NZs, this correlating with the decrease in intracellular hydrogen peroxide production. The results provide the first example of complementation of the natural enzyme function by synthetic NZs, the phenomenon which can further be used in a screening for new catalase-like nanozymes and as a fruitful tool to modify living cells by nanoparticles possessing catalytic activity and to use such modified cells as sensitive elements in cell-based biosensors., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The aim of the current research is to design alcohol oxidase-based amperometric biosensors (ABSs) using hybrid metallic nanoparticles as artificial peroxidases (PO) or PO-like nanozymes (NZs). A lot of metallic PO-like NZs were synthesized and tested with respect to their ability to substitute natural PO in solution and on amperometric electrode. The most effective PO mimetics were coupled with alcohol oxidase (AOX) on graphite electrodes (GEs) and characterized. Two types of modified GEs, namely, the AOX/nAuCePt/GE and the AOX/nFePtAu/GE show the highest sensitivities to ethanol (2600 A⋅M -1 ⋅m -2 and 1250 A⋅M -1 ⋅m -2 , respectively), low limits of detection (1.5 µM and 2.2 µM), broad linear ranges (5 - 100 µM and 12 - 120 µM), as well as satisfactory storage stabilities. The most sensitive bioelectrode AOX/nAuCePt/GE was used as ABS for ethanol determination in real samples. The practical feasibility of the constructed ABS was demonstrated by determination of ethanol in beverages, human blood and saliva., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The search for new nanoscale materials with predictable properties to target the timely and fast detection of toxic components in wastewater is one of the most promising directions of modern biosensorics. We have shown that TiO 2 nanoparticles modified with sulfur significantly improve the main operational parameters of laccase-based electrodes when compared with controls. The nanoparticle samples were labeled as TiO 2 S(0.75), TiO 2 S(1.5), and TiO 2 S(3.0), in which the numbers in parentheses refer to the quantity of H 2 SO 4 (mL) used in the synthesis. The nanoparticles and enzyme were immobilized by means of Nafion film formed on a carbon rod electrode. It was shown that the modification of Nafion film by TiO 2 or TiO 2 S(1.5) nanoparticles does not affect the size of the nanocavities and defect structure of the main polymer matrix as revealed by positron annihilation spectroscopy. It testifies that the structural-morphological difference between the film samples is rather small, and the improving of the sensor operational parameters for TiO 2 S(1.5)-based laccase electrodes is connected only with the impact of sulfur doping, but not the difference in membrane properties. The developed bioelectrodes were tested for phenol analysis in real communal wastewater samples spiked with these analytes, demonstrating the high accuracy of the assay., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Laccase is a copper-containing enzyme that does not require hydrogen peroxide as a co-substrate or additional cofactors for an enzymatic reaction. Nanomaterials of various chemical structures are usually applied to the construction of enzyme-based biosensors. Metals, metal oxides, semiconductors, and composite NPs perform various functions in electrochemical transformation schemes as a platform for the enzyme immobilization, a mediator of an electron transfer, and a signal amplifier. We describe here the development of amperometric biosensors (ABSs) based on laccase and redox-active micro/nanoparticles (hereafter-NPs), which were immobilized on a graphite electrode (GE). For this purpose, we isolated a highly purified enzyme from the fungus Trametes zonatus , and then synthesized bi- and trimetallic NPs of noble and transition metals, as well as hexacyanoferrates (HCF) of noble metals; these were layered onto the surfaces of GEs. The electroactivity of many of the NPs immobilized on the GEs was characterized by cyclic voltammetry (CV) experiments. The most effective mediators of electron transfer were selected as the platform for the development of laccase-based ABSs. As a result, a number of catechol-sensitive ABSs were constructed and characterized. The laccase/CuCo/GE was demonstrated to possess the highest sensitivity to catechol (4523 A·M -1 ·m -2 ) among the tested ABSs. The proposed ABSs may be promising for the analysis of phenolic derivatives in real samples of drinking water, wastewater, and food products.

Nanozymes (NZs) are nanoparticles that mimic the catalytic properties of natural enzymes. The present work aimed to obtain effective peroxidase mimetics (PO-like NZs), to characterize their morphological properties, estimate the kinetic parameters of NZs and evaluate the prospects of their application in analysis of ethanol. Herein, we have proposed a convenient spectrophotometric method for ethanol assay using reusable alginate beads enriched with alcohol oxidase (AO) and nanoparticles of PtCu (nPtCu) as PO-like NZs, and 3,3',5,5'-tetramethylbenzidine (TMB) as a chromogen. The linear range for the proposed nPtCu-AO/alginate beads/TMB-based method is from 0.01 mM to 0.15 mM with a limit of detection of 3.3 μM ethanol. The method is used for the quantitative determination of ethanol in alcoholic beverages. The obtained results proved to be in a good correlation with the enzymatic reference method. These results highlight the potential of the nPtCu with PO-like activity in bioanalytical applications. The proposed method, being sensitive, economical and suitable for routine and micro-volume formats, can be used in clinical diagnostics for the detection of ethanol., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Metallic nanoparticles potentially have wide practical applications in various fields of science and industry. In biosensorics, they usually act as catalysts or nanozymes (NZs) and as mediators of electron transfer. We describe here the development of amperometric biosensors (ABSs) based on purified oxidases, synthesized nanoparticles of CuCe (nCuCe), and micro/nanoporous gold (pAu), which were electro-deposited on a graphite electrode (GE). As an effective peroxidase (PO)-like NZ, nCuCe was used here as a hydrogen-peroxide-sensing platform in ABSs that were based on glucose oxidase, alcohol oxidase, methylamine oxidase, and L-arginine oxidase. At the same time, nCuCe is an electroactive mediator and has been used in laccase-based ABSs. As a result, the ABSs we constructed and characterized were based on glucose, methanol, methyl amine, L-arginine, and catechol, respectively. The developed nCuCe-based ABSs exhibited improved analytical characteristics in comparison with the corresponding PO-based ABSs. Additionally, the presence of pAu, with its extremely advanced chemo-sensing surface layer, was shown to significantly increase the sensitivities of all constructed ABSs. As an example, the bioelectrodes containing laccase/GE, laccase/nCuCe/GE, and laccase/nCuCe/pAu/GE exhibited sensitivities to catechol at 2300, 5055, and 9280 A·M -1 ·m -2 , respectively. We demonstrate here that pAu is an effective carrier of electroactive nanomaterials coupled with oxidases, which may be promising in biosensors.

Nanozymes (NZs) as stable cost-effective mimics of natural enzymes may be promising catalysts in food and environmental biotechnology, biosensors, alternative energy and medicine. The majority of known NZs are mimetics of oxidoreductases, although there are only limited data regarding mimetics of reductases. In the present research, a number of metal-based NZs were synthesized via chemical methods and screened for their antioxidant ability in solution. The most effective reductase-like Zn/Cd/Cu NZ was characterized in detail. Its antioxidant properties in comparison with several food products and Trolox, as well as substrate specificity, size and composition were studied. Zn/Cd/Cu NZ was shown to mimic preferentially selenite reductase. The amperometric sensor was constructed possessing a high sensitivity (1700 A M -1 m -2 ) and a broad linear range (16-1000 μM) for selenite ions. The possibility to apply the fabricated sensor for selenite determination in commercial mineral water has been demonstrated., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

The current review is devoted to nanozymes, i.e., nanostructured artificial enzymes which mimic the catalytic properties of natural enzymes. Use of the term "nanozyme" in the literature as indicating an enzyme is not always justified. For example, it is used inappropriately for nanomaterials bound with electrodes that possess catalytic activity only when applying an electric potential. If the enzyme-like activity of such a material is not proven in solution (without applying the potential), such a catalyst should be named an "electronanocatalyst", not a nanozyme. This paper presents a review of the classification of the nanozymes, their advantages vs. natural enzymes, and potential practical applications. Special attention is paid to nanozyme synthesis methods (hydrothermal and solvothermal, chemical reduction, sol-gel method, co-precipitation, polymerization/polycondensation, electrochemical deposition). The catalytic performance of nanozymes is characterized, a critical point of view on catalytic parameters of nanozymes described in scientific papers is presented and typical mistakes are analyzed. The central part of the review relates to characterization of nanozymes which mimic natural enzymes with analytical importance ("nanoperoxidase", "nanooxidases", "nanolaccase") and their use in the construction of electro-chemical (bio)sensors ("nanosensors").

Catalytically active nanomaterials have several advantages over their natural analogues when used as artificial enzymes (nanozymes), namely, higher stability and lower cost. Nanozymes with metallic nanocomposites are promising catalysts for biosensing applications. The aim of the current research is to construct oxidase-based bioelectrodes for food analysis using nanozymes as peroxidase mimetics. Bimetallic PtRu nanoparticles (nPtRu) coupled with alcohol oxidase (AO) and methylamine oxidase (AMO) were chosen to construct amperometric biosensors (ABSs) for primary alcohols and methylamine (MA). Both ABSs show high sensitivities (336 A·M -1 ·m -2 for the AO-ABS and 284 A·M -1 ·m -2 for the AMO-ABS), broad linear ranges (25-200 µM ethanol and 20-600 µM MA) and satisfactory storage stabilities. Practical feasibility of the constructed ABSs was demonstrated on food samples. High correlation between contents of MA and ethanol in foods determined by the ABSs and reference methods was observed., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The goal of the review is description of the main characteristics of creatinine deiminase (CDI), an important bioanalytical tool for creatinine (Crn) assay. Crn is an essential metabolite for diagnostics of kidney disfunction and some other diseases, a biomarker to control the hemodialysis procedure, as well as an important analyte for sport medicine (estimation of general physiological status of athletes). We have described the important sources for CDI isolation, cloning of the corresponding gene, the construction of microbial recombinant strains, overproducing CDI, and characteristics of the enzyme from different microorganisms. There are reviewing also the new bioanalytical methods for quantitative determination of Crn, including enzymatic ones based on using CDI., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

A novel enzymatic method of manganese (II) and cobalt (II) ions assay, based on using apo-enzyme of Mn 2+ -dependent recombinant arginase I (arginase) and 2,3-butanedione monoxime (DMO) as a chemical reagent is proposed. The principle of the method is the evaluation of the activity of L-arginine-hydrolyzing of arginase holoenzyme after the specific binding of Mn 2+ or Co 2+ with apo-arginase. Urea, which is the product of enzymatic hydrolysis of L-arginine (Arg), reacts with DMO and the resulted compound is detected by both fluorometry and visual spectrophotometry. Thus, the content of metal ions in the tested samples can be determined by measuring the level of urea generated after enzymatic hydrolysis of Arg by reconstructed arginase holoenzyme in the presence of tested metal ions. The linearity range of the fluorometric apo-arginase-DMO method in the case of Mn 2+ assay is from 4pM to 1.10nM with a limit of detection of 1pM Mn 2+ , whereas the linearity range of the present method in the case of Co 2+ assay is from 8pM to 45nM with a limit of detection of 2.5pM Co 2+ . The proposed method being highly sensitive, selective, valid and low-cost, may be useful to monitor Mn 2+ and Co 2+ content in clinical laboratories, food industry and environmental control service., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The enzymes of l-arginine (further - Arg) metabolism are promising tools for elaboration of selective methods for quantitative Arg analysis. In our study we propose an enzymatic method for Arg assay based on fluorometric monitoring of ammonia, a final product of Arg splitting by human liver arginase I (further - arginase), isolated from the recombinant yeast strain, and commercial urease. The selective analysis of ammonia (at 415nm under excitation at 360nm) is based on reaction with o-phthalaldehyde (OPA) in the presence of sulfite in alkali medium: these conditions permit to avoid the reaction of OPA with any amino acid. A linearity range of the fluorometric arginase-urease-OPA method is from 100nM to 6μМ with a limit of detection of 34nM Arg. The method was used for the quantitative determination of Arg in the pooled sample of blood serum. The obtained results proved to be in a good correlation with the reference enzymatic method and literature data. The proposed arginase-urease-OPA method being sensitive, economical, selective and suitable for both routine and micro-volume formats, can be used in clinical diagnostics for the simultaneous determination of Arg as well as urea and ammonia in serum samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A highly selective and sensitive enzymatic method for the quantitative determination of L-arginine (Arg) has been developed. The method is based on the use of recombinant bacterial arginine deiminase (ADI) isolated from the cells of a recombinant strain Escherichia coli and o-phthalaldehyde (OPA) as a chemical reagent. Ammonia, the product of the enzymatic digestion of Arg by ADI, reacts with OPA and forms in the presence of sulfite a product, which can be detected by spectrophotometry (S) and fluorometry (F). The linear concentration range for Arg assay in the final reaction mixture varies for ADI-OPA-F variant of the method from 0.35 μM to 24 μM with the detection limit of 0.25 μM. For ADI-OPA-S variant of the assay, the linearity varies from 0.7 μM to 50 μM with the detection limit of 0.55 μM. The new method was tested on real samples of wines and juices. A high correlation (R=0.978) was shown for the results obtained with the proposed and the reference enzymatic method., (Copyright © 2016 Elsevier Ltd. All rights reserved.)