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Design of bioluminescent biosensors for assessing contamination of complex matrices / E. N. Esimbekova, V. P. Kalyabina, K. V. Kopylova [et al.] // Talanta. - 2021. - Vol. 233. - Ст. 122509, DOI 10.1016/j.talanta.2021.122509. - Cited References:87. - The reported study was funded by Russian Foundation for Basic Research, Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science, to the research project No. 20-44-242001 and Ministry of Science and Higher Education of Russian Federation No. FSRZ-2020-0006. . - ISSN 0039-9140. - ISSN 1873-3573

РУБ Chemistry, Analytical
Рубрики:
SAMPLE PREPARATION
   PESTICIDES
   FOOD
   BIOMOLECULES
   SENSITIVITY
Кл.слова (ненормированные):
Bioluminescent biosensor -- Enzyme inhibition-based assay -- Complex -- matrices -- Pesticides -- Heavy metals
Аннотация: The presence of potentially toxic xenobiotics in complex matrices has become rather the rule than the exception. Therefore, there is a need for highly sensitive inexpensive techniques for analyzing environmental and food matrices for toxicants. Enzymes are selectively sensitive to various toxic compounds, and, thus, they can be used as the basis for detection of contaminants in complex matrices. There are, however, a number of difficulties associated with the analysis of complex matrices using enzyme assays, including the necessity to take into account properties and effects of the natural components of the test media for accurate interpretation of results. The present study describes the six-stage procedure for designing new enzyme sensors intended for assessing the quality of complex matrices. This procedure should be followed both to achieve the highest possible sensitivity of the biosensor to potentially toxic substances and to minimize the effect of the uncontaminated components of complex mixtures on the activity of the biosensor. The proposed strategy has been tested in designing a bioluminescent biosensor for integrated rapid assessment of the safety of fruits and vegetables. The biosensor is based on the coupled enzyme system NAD(P)H:FMN-oxidoreductase and luciferase as the biorecognition element. The study describes methods and techniques for attaining the desired result in each stage. The proposed six-stage procedure for designing bioluminescent enzyme biosensors can be used to design the enzymatic biosensors based on other enzymes.

WOS
Держатели документа:
Siberian Fed Univ, 79 Svobodny Prospect, Krasnoyarsk 660041, Russia.
Inst Biophys SB RAS, 50-50 Akademgorodok, Krasnoyarsk 660036, Russia.

Доп.точки доступа:
Esimbekova, Elena N.; Kalyabina, Valeriya P.; Kopylova, Kseniya, V; Torgashina, Irina G.; Kratasyuk, Valentina A.; Kopylova, Kseniya; Esimbekova, Elena; Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR); Government of Krasnoyarsk Territory; Krasnoyarsk Regional Fund of Science [20-44-242001]; Ministry of Science and Higher Education of Russian Federation [FSRZ-2020-0006]

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Design of bioluminescent biosensors for assessing contamination of complex matrices / E. N. Esimbekova, V. P. Kalyabina, K. V. Kopylova [et al.] // Talanta. - 2021. - Vol. 233. - Ст. 122509, DOI 10.1016/j.talanta.2021.122509. - Cited By :1 . - ISSN 0039-9140
Кл.слова (ненормированные):
Bioluminescent biosensor -- Complex matrices -- Enzyme inhibition-based assay -- Heavy metals -- Pesticides
Аннотация: The presence of potentially toxic xenobiotics in complex matrices has become rather the rule than the exception. Therefore, there is a need for highly sensitive inexpensive techniques for analyzing environmental and food matrices for toxicants. Enzymes are selectively sensitive to various toxic compounds, and, thus, they can be used as the basis for detection of contaminants in complex matrices. There are, however, a number of difficulties associated with the analysis of complex matrices using enzyme assays, including the necessity to take into account properties and effects of the natural components of the test media for accurate interpretation of results. The present study describes the six-stage procedure for designing new enzyme sensors intended for assessing the quality of complex matrices. This procedure should be followed both to achieve the highest possible sensitivity of the biosensor to potentially toxic substances and to minimize the effect of the uncontaminated components of complex mixtures on the activity of the biosensor. The proposed strategy has been tested in designing a bioluminescent biosensor for integrated rapid assessment of the safety of fruits and vegetables. The biosensor is based on the coupled enzyme system NAD(P)H:FMN-oxidoreductase and luciferase as the biorecognition element. The study describes methods and techniques for attaining the desired result in each stage. The proposed six-stage procedure for designing bioluminescent enzyme biosensors can be used to design the enzymatic biosensors based on other enzymes. © 2021 Elsevier B.V.

Scopus
Держатели документа:
Siberian Federal University, 79 Svobodny Prospect, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, 50/50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Esimbekova, E. N.; Kalyabina, V. P.; Kopylova, K. V.; Torgashina, I. G.; Kratasyuk, V. A.

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РУБ Biotechnology & Applied Microbiology + Engineering, Environmental
Рубрики:
FLUORIDE
   BIOASSAYS
   POLLUTION
   METALS
   WATER
Кл.слова (ненормированные):
Urbostratozems -- Soil pollution -- Industrial contamination -- Bioassay -- Bioluminescence
Аннотация: There is a need for rapid simple and informative environmental assessment methods. The present investigation is aimed at assessing the possibility of using the combined enzyme system of luminescent bacteria: NAD(P)H:FMN-oxidoreductase + luciferase (Red + Luc) for predicting the potential toxicity of industrial urbostratozems sampled in the city of Krasnoyarsk. Three groups of urbostratozems polluted with fluorine, arsenic and lead, were tested by the methods of chemical analysis and enzymatic bioassay. Only the assessment of the arsenic-contaminated soil samples showed the dependence between the reduced activity of the enzyme system and the arsenic concentration variations. The results reveal that the sensitivity of the Red + Luc enzyme system to the soil pollutants depends on the properties of the studied soil samples. Moreover, the solubility of lead in the soil samples affects the accuracy of the enzymatic bioassays for soil toxicity testing. The results of the enzymatic bioassay of the fluoride-contaminated soil samples are ambiguous. The obtained data show the relevance of the sample preparation during integral bioassays. In addition, soil properties should be taken into account as well. The current study emphasizes the importance of conducting chemical and biological testing as a combined set to obtain comprehensive information about the anthropogenic load. (C) 2021 Elsevier B.V. All rights reserved.

WOS
Держатели документа:
Siberian Fed Univ, Dept Biophys, 79 Svobodny St, Krasnoyarsk 660041, Russia.
Russian Acad Sci, Krasnoyarsk Agr Res Inst, Fed Res Ctr Krasnoyarsk Sci Ctr Siberian Branch, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Dept Aquat & Terr Ecosyst, Krasnoyarsk 660041, Russia.
Russian Acad Sci, Photobiol Lab, Inst Biophys, Fed Res Ctr `Krasnoyarsk Sci Ctr Siberian Branch, Krasnoyarsk 660036, Russia.

Доп.точки доступа:
Kolosova, Elizaveta M.; Sutormin, Oleg S.; Stepanova, L. V.; Shpedt, Aleksandr A.; Rimatskaya, N. V.; Sukovataya, Irina E.; Kratasyuk, Valentina A.; Russian Foundation for Basic Research, the Government of the Krasnoyarsk Region, Russia; Krasnoyarsk Regional Foundation for Supporting Scientific and Technological Activities, Russia [18-47-240005]; Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]

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Bioluminescent enzyme inhibition-based assay for the prediction of toxicity of pollutants in urban soils / E. M. Kolosova, O. S. Sutormin, L. V. Stepanova [et al.] // Environ. Technol. Innov. - 2021. - Vol. 24. - Ст. 101842, DOI 10.1016/j.eti.2021.101842 . - ISSN 2352-1864
Кл.слова (ненормированные):
Bioassay -- Bioluminescence -- Industrial contamination -- Soil pollution -- Urbostratozems -- Arsenic -- Chemical analysis -- Enzyme activity -- Enzyme inhibition -- Fluorine compounds -- Soil surveys -- Soil testing -- Soils -- Toxicity -- Arsenic concentration -- Chemical and biologicals -- Comprehensive information -- Contaminated soils -- Environmental assessment methods -- Enzymatic bioassays -- Luminescent bacteria -- Sample preparation -- Soil pollution
Аннотация: There is a need for rapid simple and informative environmental assessment methods. The present investigation is aimed at assessing the possibility of using the combined enzyme system of luminescent bacteria: NAD(P)H:FMN-oxidoreductase + luciferase (Red + Luc) for predicting the potential toxicity of industrial urbostratozems sampled in the city of Krasnoyarsk. Three groups of urbostratozems polluted with fluorine, arsenic and lead, were tested by the methods of chemical analysis and enzymatic bioassay. Only the assessment of the arsenic-contaminated soil samples showed the dependence between the reduced activity of the enzyme system and the arsenic concentration variations. The results reveal that the sensitivity of the Red + Luc enzyme system to the soil pollutants depends on the properties of the studied soil samples. Moreover, the solubility of lead in the soil samples affects the accuracy of the enzymatic bioassays for soil toxicity testing. The results of the enzymatic bioassay of the fluoride-contaminated soil samples are ambiguous. The obtained data show the relevance of the sample preparation during integral bioassays. In addition, soil properties should be taken into account as well. The current study emphasizes the importance of conducting chemical and biological testing as a combined set to obtain comprehensive information about the anthropogenic load. © 2021 Elsevier B.V.

Scopus
Держатели документа:
Department of Biophysics, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Krasnoyarsk Agricultural Research Institute, Federal Research Center ‘Krasnoyarsk Science Center Siberian Branch of the Russian Academy of Sciences’, Krasnoyarsk, 660036, Russian Federation
Department of Aquatic and Terrestrial Ecosystems, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Photobiology Laboratory, Institute of Biophysics, Federal Research Center ‘Krasnoyarsk Science Center Siberian Branch of the Russian Academy of Sciences’, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Kolosova, E. M.; Sutormin, O. S.; Stepanova, L. V.; Shpedt, A. A.; Rimatskaya, N. V.; Sukovataya, I. E.; Kratasyuk, V. A.

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Effects of Modified Magnetite Nanoparticles on Bacterial Cells and Enzyme Reactions / L. S. Bondarenko, E. S. Kovel, K. A. Kydralieva [et al.] // Nanomaterials. - 2020. - Vol. 10, Is. 8. - Ст. 1499, DOI 10.3390/nano10081499. - Cited References:83. - This research was funded by the Russian Foundation for Basic Research (#19-315-50048, #19-33-90149, and #18-29-19003). . - ISSN 2079-4991

РУБ Nanoscience & Nanotechnology + Materials Science, Multidisciplinary
Рубрики:
NATURAL ORGANIC-MATTER
HUMIC-ACID
DETOXIFICATION PROCESSES
Кл.слова (ненормированные):
magnetite nanoparticles -- humic acids-coated magnetite nanoparticles -- silica-coated magnetite nanoparticles -- zeta potential -- hydrodynamic -- diameter -- toxicity -- bioluminescence -- bacterial assay -- enzymatic assay -- oxidative stress -- Photobacterium phosphoreum -- NADH -- FMN-oxidoreductase -- luciferase
Аннотация: Current paper presents biological effects of magnetite nanoparticles (MNPs). Relations of MNP' characteristics (zeta-potential and hydrodynamic diameters) with effects on bacteria and their enzymatic reactions were the main focus.Photobacterium phosphoreumand bacterial enzymatic reactions were chosen as bioassays. Three types of MNPs were under study: bare Fe3O4, Fe(3)O(4)modified with 3-aminopropyltriethoxysilane (Fe3O4/APTES), and humic acids (Fe3O4/HA). Effects of the MNPs were studied at a low concentration range (< 2 mg/L) and attributed to availability and oxidative activity of Fe3+, high negative surface charge, and low hydrodynamic diameter of Fe3O4/HA, as well as higher Fe(3+)content in suspensions of Fe3O4/HA. Low-concentration suspensions of bare Fe(3)O(4)provided inhibitory effects in both bacterial and enzymatic bioassays, whereas the MNPs with modified surface (Fe3O4/APTES and Fe3O4/HA) did not affect the enzymatic activity. Under oxidative stress (i.e., in the solutions of model oxidizer, 1,4-benzoquinone), MNPs did not reveal antioxidant activity, moreover, Fe3O4/HA demonstrated additional inhibitory activity. The study contributes to the deeper understanding of a role of humic substances and silica in biogeochemical cycling of iron. Bioluminescence assays, cellular and enzymatic, can serve as convenient tools to evaluate bioavailability of Fe(3+)in natural dispersions of iron-containing nanoparticles, e.g., magnetite, ferrihydrite, etc.

WOS
Держатели документа:
Natl Res Univ, Moscow Aviat Inst, Moscow 125993, Russia.
FRC KSC SB RAS, Inst Phys SB RAS, Krasnoyarsk 660036, Russia.
FRC KSC SB RAS, Inst Biophys SB RAS, Krasnoyarsk 660036, Russia.
RAS, Moscow Inst Problems Chem Phys, Chernogolovka 142432, Moscow Region, Russia.
Univ Szeged, H-6720 Szeged, Hungary.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Bondarenko, Lyubov S.; Kovel, Ekaterina S.; Kydralieva, Kamila A.; Dzhardimalieva, Gulzhian, I; Illes, Erzsebet; Tombacz, Etelka; Kicheeva, Arina G.; Kudryasheva, Nadezhda S.; Dzhardimalieva, Gulzhian; Kudryasheva, Nadezhda; Kovel, Ekaterina; Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [19-315-50048, 19-33-90149, 18-29-19003]

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Effects of modified magnetite nanoparticles on bacterial cells and enzyme reactions / L. S. Bondarenko, E. S. Kovel, K. A. Kydralieva [et al.] // Nanomaterials. - 2020. - Vol. 10, Is. 8. - Ст. 1499. - P1-20, DOI 10.3390/nano10081499 . - ISSN 2079-4991
Кл.слова (ненормированные):
Bacterial assay -- Bioluminescence -- Enzymatic assay -- Humic acids-coated magnetite nanoparticles -- Hydrodynamic diameter -- Luciferase -- Magnetite nanoparticles -- NADH:FMN-oxidoreductase -- Oxidative stress -- Photobacterium phosphoreum -- Silica-coated magnetite nanoparticles -- Toxicity -- Zeta potential
Аннотация: Current paper presents biological effects of magnetite nanoparticles (MNPs). Analyzing effects of MNP’ characteristics (zeta-potential and hydrodynamic diameters) on bacteria and their enzyme reactions was the main focus. Photobacterium phosphoreum and bacterial enzymatic reactions were chosen as bioassays. Three types of MNPs were under study: bare Fe3O4, Fe3O4 modified with 3-aminopropyltriethoxysilane (Fe3O4/APTES), and humic acids (Fe3O4/HA). Effects of the MNPs were studied at a low concentration range (< 2 mg/L) and attributed to availability and oxidative activity of Fe3+, high negative surface charge, and low hydrodynamic diameter of Fe3O4/HA, as well as higher Fe3+ content in suspensions of Fe3O4/HA. Low-concentration suspensions of bare Fe3O4 provided inhibitory effects in both bacterial and enzymatic bioassays, whereas the MNPs with modified surface (Fe3O4/APTES and Fe3O4/HA) did not affect the enzymatic activity. Under oxidative stress (i.e., in the solutions of model oxidizer, 1,4-benzoquinone), MNPs did not reveal antioxidant activity, moreover, Fe3O4/HA demonstrated additional inhibitory activity. The study contributes to the deeper understanding of a role of humic substances and silica in biogeochemical cycling of iron. Bioluminescence assays, cellular and enzymatic, can serve as convenient tools to evaluate bioavailability of Fe3+ in natural dispersions of iron-containing nanoparticles, e.g., magnetite, ferrihydrite, etc. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

Scopus
Держатели документа:
Moscow Aviation Institute (National Research University), Moscow, 125993, Russian Federation
Institute of Physics SB RAS, FRC KSC SB RAS, Krasnoyarsk, 660036, Russian Federation
Institute of Biophysics SB RAS, FRC KSC SB RAS, Krasnoyarsk, 660036, Russian Federation
Institute of Problems of Chemical Physics RAS, Moscow Region, Chernogolovka, 142432, Russian Federation
University of Szeged, Szeged, H-6720, Hungary
Siberian Federal University, Krasnoyarsk, 660041, Russian Federation

Доп.точки доступа:
Bondarenko, L. S.; Kovel, E. S.; Kydralieva, K. A.; Dzhardimalieva, G. I.; Illes, E.; Tombacz, E.; Kicheeva, A. G.; Kudryasheva, N. S.

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Gelatin and Starch: What Better Stabilizes the Enzyme Activity? / E. N. Esimbekova, A. E. Govorun, V. I. Lonshakova-Mukina, V. A. Kratasyuk // Dokl. Biol. Sci. - 2020. - Vol. 491, Is. 1. - P43-46, DOI 10.1134/S0012496620020039 . - ISSN 0012-4966
Кл.слова (ненормированные):
butyrylcholinesterase -- enzyme stabilization -- gelatin -- luciferase -- NAD(P)H:FMN oxidoreductase -- starch -- thermal inactivation of enzymes
Аннотация: Abstract: The regularities of the functioning of a number of enzymes in a viscous environment created by natural polymers, starch and gelatin are examined. Based on the analysis of kinetic curves of thermal inactivation, mechanisms of thermal inactivation of enzymes in a viscous microenvironment are proposed. Using the example of butyrylcholinesterase, NAD(P)H:FMN oxidoreductase, and coupled system of the luminous bacteria (NAD(P)H:FMN oxidoreductase + luciferase), the conditions, under which starch and gelatin have a stabilizing effect on enzyme activity during storage and exposure to various physical and chemical environmental factors, were found. A significant increase in the stabilizing effect is achieved by eliminating water during drying the enzyme preparations immobilized in starch and gelatin polymer gels. © 2020, Pleiades Publishing, Ltd.

Scopus
Держатели документа:
Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, Krasnoyarsk, 660036, Russian Federation
Siberian Federal University, Krasnoyarsk, 660041, Russian Federation

Доп.точки доступа:
Esimbekova, E. N.; Govorun, A. E.; Lonshakova-Mukina, V. I.; Kratasyuk, V. A.

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Enzymatic responses to low-intensity radiation of tritium / T. V. Rozhko, E. V. Nemtseva, M. V. Gardt [et al.] // Int. J. Mol. Sci. - 2020. - Vol. 21, Is. 22. - Ст. 8464. - P1-15, DOI 10.3390/ijms21228464 . - ISSN 1661-6596
Кл.слова (ненормированные):
Bacterial luciferase -- Enzymes -- Fluorescent protein -- Hormesis -- Low-dose radiation -- Oxidoreductase -- Tritium
Аннотация: The present study considers a possible role of enzymatic reactions in the adaptive response of cells to the beta-emitting radionuclide tritium under conditions of low-dose exposures. Effects of tritiated water (HTO) on the reactions of bacterial luciferase and NAD(P)H:FMN-oxidoreductase, as well as a coupled system of these two reactions, were studied at radioactivity concentrations ? 200 MBq/L. Additionally, one of the simplest enzymatic reactions, photobiochemical proton transfer in Coelenteramide-containing Fluorescent Protein (CLM-FP), was also investigated. We found that HTO increased the activity of NAD(P)H:FMN-oxidoreductase at the initial stage of its reaction (by up to 230%); however, a rise of luciferase activity was moderate (<20%). The CLM-FP samples did not show any increase in the rate of the photobiochemical proton transfer under the exposure to HTO. The responses of the enzyme systems were compared to the ‘hormetic’ response of luminous marine bacterial cells studied earlier. We conclude that (1) the oxidoreductase reaction contributes significantly to the activation of the coupled enzyme system and bacterial cells by tritium, and (2) an increase in the organization level of biological systems promotes the hormesis phenomenon. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.

Scopus
Держатели документа:
Department of Medical and Biological Physics, Krasnoyarsk State Medical Academy, Krasnoyarsk, 660022, Russian Federation
Biophysics Department, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, FRC KSC SB RAS, Krasnoyarsk, 660036, Russian Federation
Department of Chemistry, Moscow State University, Moscow, 119991, Russian Federation

Доп.точки доступа:
Rozhko, T. V.; Nemtseva, E. V.; Gardt, M. V.; Raikov, A. V.; Lisitsa, A. E.; Badun, G. A.; Kudryasheva, N. S.

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Enzymatic Responses to Low-Intensity Radiation of Tritium / T. V. Rozhko, E. V. Nemtseva, M. V. Gardt [et al.] // Int. J. Mol. Sci. - 2020. - Vol. 21, Is. 22. - Ст. 8464, DOI 10.3390/ijms21228464. - Cited References:59. - This work was supported by RFBR-Krasnoyarsk Regional Foundation N 18-44-240004, 18-44-242002. . - ISSN 1422-0067

РУБ Biochemistry & Molecular Biology + Chemistry, Multidisciplinary
Рубрики:
LUMINOUS MARINE-BACTERIA
IONIZING-RADIATION
DISCHARGED-OBELIN
Кл.слова (ненормированные):
hormesis -- low-dose radiation -- tritium -- enzymes -- bacterial luciferase -- oxidoreductase -- fluorescent protein
Аннотация: The present study considers a possible role of enzymatic reactions in the adaptive response of cells to the beta-emitting radionuclide tritium under conditions of low-dose exposures. Effects of tritiated water (HTO) on the reactions of bacterial luciferase and NAD(P)H:FMN-oxidoreductase, as well as a coupled system of these two reactions, were studied at radioactivity concentrations <= 200 MBq/L. Additionally, one of the simplest enzymatic reactions, photobiochemical proton transfer in Coelenteramide-containing Fluorescent Protein (CLM-FP), was also investigated. We found that HTO increased the activity of NAD(P)H:FMN-oxidoreductase at the initial stage of its reaction (by up to 230%); however, a rise of luciferase activity was moderate (<20%). The CLM-FP samples did not show any increase in the rate of the photobiochemical proton transfer under the exposure to HTO. The responses of the enzyme systems were compared to the 'hormetic' response of luminous marine bacterial cells studied earlier. We conclude that (1) the oxidoreductase reaction contributes significantly to the activation of the coupled enzyme system and bacterial cells by tritium, and (2) an increase in the organization level of biological systems promotes the hormesis phenomenon.

WOS
Держатели документа:
Krasnoyarsk State Med Acad, Dept Med & Biol Phys, Krasnoyarsk 660022, Russia.
Siberian Fed Univ, Biophys Dept, Krasnoyarsk 660041, Russia.
RAS, Inst Biophys, SB, FRC,KSC, Krasnoyarsk 660036, Russia.
Moscow MV Lomonosov State Univ, Dept Chem, Moscow 119991, Russia.

Доп.точки доступа:
Rozhko, Tatiana V.; Nemtseva, Elena V.; Gardt, Maria V.; Raikov, Alexander V.; Lisitsa, Albert E.; Badun, Gennadii A.; Kudryasheva, Nadezhda S.; Nemtseva, Elena; Kudryasheva, Nadezhda; Rozko, Tat'ana; Lisitsa, Albert; RFBR-Krasnoyarsk Regional Foundation [N 18-44-240004, 18-44-242002]

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10.

A noninvasive and qualitative bioluminescent assay for express diagnostics of athletes' responses to physical exertion / V. A. Kratasyuk, L. V. Stepanova, R. Ranjan [et al.] // Luminescence. - 2020, DOI 10.1002/bio.3954. - Cited References:33. - The Ministry of Science and Higher Education of the Russian Federation, Grant/Award Number: FSRZ-2020-0006; Krasnoyarsk Regional Foundation of Science, Grant/Award Number: KF-537 . - Article in press. - ISSN 1522-7235. - ISSN 1522-7243

РУБ Chemistry, Analytical
Рубрики:
SALIVARY BIOMARKERS
EXERCISE
Кл.слова (ненормированные):
athletes -- BLuc‐ -- Red coupled enzyme system -- catalase activity -- saliva -- training load
Аннотация: Upcoming professional sports authorities seek rapid noninvasive biosensing tools for regular monitoring of athletes' physiological states. The analysis of saliva through luminescence-based biosensors has been perceived as a suitable candidate for such purposes. The present study reports a qualitative bioluminescence assay based on a coupled enzyme system that consists of bacterial luciferase (BLuc) and nicotinamide adenine dinucleotide (NADH):flavin mononucleotide (FMN) oxidoreductase (Red), BLuc-Red, for the express diagnostics of athletes' stress levels before and after physical exertion. The volunteers who participated in the study were grouped as freestyle wrestlers and students who adapted to different levels of physical activities. Under physical exertion modelling conditions, the influence of participant saliva on BLuc-Red catalyzed light emission was investigated. Results showed a significant increase in residual luminescence (I-exp, mean maximum bioluminescence intensity of the experimental measurement (I-exp); I-c, luminescence intensity in control; I-exp/I-c, %) values for participants in the wrestler group while a decrease in the student group (P < 0.05). Such contrasting residual luminescence values in both groups were found to be dependent on the catalase activity of saliva. The proposed bioluminescence assay can be utilized as a potential nonspecific biosensing tool for determining the physical state of athletes under high loads.

WOS
Держатели документа:
Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, Dept Biophys, Svobodny Prospect 79, Krasnoyarsk 660041, Russia.
Inst Biophys SB RAS, Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Akademgorodok 50-50, Akademgorodok, Russia.
Krasnoyarsk State Med Univ, Minist Hlth Russian Federat, Av Partizan Zheleznyak 1, Krasnoyarsk, Russia.
Krasnoyarsk Matern & Childhood Protect Ctr, Kirenskogo St 2a, Krasnoyarsk, Russia.
Siberian Fed Univ, Sch Nonferrous Met & Mat Sci, Svobodny Prospect 79, Krasnoyarsk, Russia.
Sci Res Inst Med Problems North, Av Partizan Zheleznyak 3g, Krasnoyarsk, Russia.

Доп.точки доступа:
Kratasyuk, Valentina A.; Stepanova, Lyudmila, V; Ranjan, Rajeev; Sutormin, Oleg S.; Pande, Shubhra; Zhukova, Galina, V; Miller, Olga M.; Maznyak, Natalya, V; Kolenchukova, Oksana A.; Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]; Krasnoyarsk Regional Foundation of Science [KF-537]

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11.

Functional divergence between evolutionary-related LuxG and Fre oxidoreductases of luminous bacteria / A. A. Deeva [et al.] // Proteins. - 2019. - Vol. 87, Is. 9. - P723-729, DOI 10.1002/prot.25696. - Cited References:39. - The Russian Foundation for Basic Research and Krasnoyarsk Region Science and Technology Support Fund, Grant/Award Number: 18-44-243009; Ministry of Education and Science of the Russian Federation, Grant/Award Numbers: 0356-2019-0019, 6.7734.2017 . - ISSN 0887-3585. - ISSN 1097-0134

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
ESCHERICHIA-COLI
FLAVIN OXIDOREDUCTASE
CRYSTAL-STRUCTURE
Кл.слова (ненормированные):
bacterial bioluminescence -- Fre -- functional divergence -- gene duplication -- LuxG -- NAD(P)H -- flavin-oxidoreductase
Аннотация: In luminous bacteria NAD(P)H:flavin-oxidoreductases LuxG and Fre, there are homologous enzymes that could provide a luciferase with reduced flavin. Although Fre functions as a housekeeping enzyme, LuxG appears to be a source of reduced flavin for bioluminescence as it is transcribed together with luciferase. This study is aimed at providing the basic conception of Fre and LuxG evolution and revealing the peculiarities of the active site structure resulted from a functional variation within the oxidoreductase family. A phylogenetic analysis has demonstrated that Fre and LuxG oxidoreductases have evolved separately after the gene duplication event, and consequently, they have acquired changes in the conservation of functionally related sites. Namely, different evolutionary rates have been observed at the site responsible for specificity to flavin substrate (Arg 46). Also, Tyr 72 forming a part of a mobile loop involved in FAD binding has been found to be conserved among Fre in contrast to LuxG oxidoreductases. The conservation of different amino acid types in NAD(P)H binding site has been defined for Fre (arginine) and LuxG (proline) oxidoreductases.

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Держатели документа:
Siberian Fed Univ, Lab Bioluminescent Biotechnol, Svobodny Prosp 79, Krasnoyarsk 660041, Russia.
RAS, Inst Cell Biophys, Mech Cell Genome Functioning Lab, Pushchino, Moscow Region, Russia.
State Inst Informat Technol & Telecommun SIIT & T, Dept Appl Res Informatizat, Moscow, Russia.
RAS, Fed Res Ctr, Krasnoyarsk Sci Ctr SB, Lab Photobiol,Inst Biophys SB, Krasnoyarsk, Russia.

Доп.точки доступа:
Deeva, Anna A.; Zykova, Evgenia A.; Nemtseva, Elena V.; Kratasyuk, Valentina A.; Nemtseva, Elena; Russian Foundation for Basic Research [18-44-243009]; Ministry of Education and Science of the Russian Federation [0356-2019-0019, 6.7734.2017]; Krasnoyarsk Region Science and Technology [18-44-243009]

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12.

NAD(P)H:FMN-Oxidoreductase Functioning Under Macromolecular Crowding: In Vitro Modeling / A. E. Govorun, E. N. Esimbekova, V. A. Kratasyuk // Doklad. Biochem. Biophys. - 2019. - Vol. 486, Is. 1. - P213-215, DOI 10.1134/S160767291903013X . - ISSN 1607-6729
Аннотация: The functioning of NAD(P)H:FMN‑oxidoreductase (Red) from Vibrio fischeri under conditions of macromolecular crowding (MMC) simulated in vitro by adding biopolymers (starch and gelatin) was studied. The dissociation rate constants and the activation energies of dissociation of Red to the subunits were calculated, and the process of denaturation of Red was analyzed. It is shown that the functioning of Red both under conditions of MMC and in diluted solutions is the same. This result refutes the common belief that the native conformation of enzymes in vivo is stabilized due to MMC as compared to the in vitro conditions. © 2019, Pleiades Publishing, Ltd.

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Держатели документа:
Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Govorun, A. E.; Esimbekova, E. N.; Kratasyuk, V. A.

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13.

Principles for Construction of Bioluminescent Enzyme Biotests for Analysis of Complex Media / V. P. Kalyabina [et al.] // Dokl. Biochem. Biophys. - 2019. - Vol. 485, Is. 1. - P107-110, DOI 10.1134/S1607672919020042. - Cited References:10. - The study was supported by the Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science and RFBR (project no. 18-44-242003). . - ISSN 1607-6729. - ISSN 1608-3091

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
VEGETABLES
ELEMENTS
Аннотация: In this study, we formulated the principles of designing bioluminescent enzyme tests for assessing the quality of complex media, which consist in providing the maximum sensitivity to potentially toxic chemicals at a minimal impact of uncontaminated complex media. The developed principles served as a basis for designing a new bioluminescent method for an integrated rapid assessment of chemical safety of fruits and vegetables, which is based on using the luminous bacteria enzymes (NAD(P)H:FMN oxidoreductase and luciferase) as a test system.

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Держатели документа:
Siberian Fed Univ, Krasnoyarsk, Russia.
Russian Acad Sci, Siberian Branch, Inst Biophys, Krasnoyarsk, Russia.

Доп.точки доступа:
Kalyabina, V. P.; Esimbekova, E. N.; Torgashina, I. G.; Kopylova, K. V.; Kratasyuk, V. A.; Government of Krasnoyarsk Territory; Krasnoyarsk Regional Fund of Science; RFBR [18-44-242003]

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14.

Towards biological quantity theory for nominal property metrology in polyenzymatic devices with living cells / P. I. Belobrov, A. A. Evstrapov, E. N. Esimbekova [et al.] // JOINT IMEKO TC1-TC7-TC13-TC18 SYMPOSIUM : IOP PUBLISHING LTD, 2019. - Vol. 1379: Joint IMEKO TC1-TC7-TC13-TC18 Symposium (JUL 02-05, 2019, St Petersburg, RUSSIA). - Ст. 012036. - (Journal of Physics Conference Series), DOI 10.1088/1742-6596/1379/1/012036. - Cited References:19 . -

РУБ Instruments & Instrumentation + Mathematics, Applied + Physics, Applied

Аннотация: Here we discuss the concepts of "biological quantity" and "nominal property" within the framework of the problem of biological measurements based on new specific results of biological analysis using a microfluidic platform and chips developed by our team earlier. It was shown that based on different microfluidic platforms it is possible to develop chips with a polyenzymatic bioluminescent system NAD(P)H:FMN-oxidoreductase-luciferase (Red + Luc), which can be used in various areas of biological analysis. Thus, disposable microfluidic chips with Red + Luc system suitable for field and indoor use were developed using continuous flow microfluidic platform. The use of droplet-based microfluidic platform allowed to develop microfluidic chips with Red + Luc system for long-term continuous measurements of water samples, for example, in places of waste disposal by industrial enterprises. The reference for comparing different biological quantities with each other in the proposed system was a photodetector, which converted non-numeric values and nominal properties recorded by a biological module Red + Luc into numerical variables. Such a reference was implemented as a portable luminometer based on silicon photomultiplier. The results allow to perform other biological measurements and to start the discussion of modern biological concepts in the language of biological measures.

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Держатели документа:
Siberian Fed Univ, Lab Bioluminescent Biotechnol, Krasnoyarsk 660041, Russia.
RAS, Inst Analyt Instrumentat, St Petersburg 190103, Russia.
SB RAS, Lab Photobiol, Inst Biophys, Krasnoyarsk 660036, Russia.
SB RAS, Krasnoyarsk Sci Ctr, Fed Res Ctr, Lab Digital Controlled Drugs & Theranost, Krasnoyarsk 660036, Russia.
Krasnoyarsk State Med Univ, Res Inst Mol Med & Pathobiochem, Krasnoyarsk 660022, Russia.

Доп.точки доступа:
Belobrov, P., I; Evstrapov, A. A.; Esimbekova, E. N.; Denisov, I. A.; Lukyanenko, K. A.; Osipova, E. D.; Yakimov, A. S.; Belobrov, Peter I; Yakimov, Anton

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15.

Set of Enzymatic Bioassays for Assessment of Soil Contamination / E. M. Kolosova, O. S. Sutormin, E. N. Esimbekova [et al.] // Dokl. Biol. Sci. - 2019. - Vol. 489, Is. 1. - P165-168, DOI 10.1134/S0012496619060024 . - ISSN 1608-3105
Аннотация: A concept of the comprehensive assessment of soil contamination is proposed. According to it, the conclusion regarding the presence of toxic substances in the analyzed sample is based on the inhibition of enzymatic reactions responsible for various functions of a living organism, such as luminescence, respiration, etc. These functions are taken as test functions in classical bioassays with the use of living objects (luminous bacteria, daphnia, algae, and others). The regularities of the impact of different classes of toxicants on the activity of particular enzymes or coupled oligo-enzyme chains have been established. These enzyme reactions are selected as potential test objects: markers of contamination. Three enzyme systems with the maximal sensitivity to different classes of toxicants have been chosen for the set of enzymatic bioassays: butyrylcholinesterase, NAD(P)H:FMN-oxidoreductase + luciferase, and lactate dehydrogenase + NAD(P)H:FMN-oxidoreductase + luciferase. The possibility to use enzymes instead of living organisms in the bioassay of natural complex systems has been shown.

Scopus
Держатели документа:
Siberian Federal University, Krasnoyarsk, Russian Federation
Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Kolosova, E. M.; Sutormin, O. S.; Esimbekova, E. N.; Lonshakova-Mukina, V. I.; Kratasyuk, V. A.

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16.

Towards biological quantity theory for nominal property metrology in polyenzymatic devices with living cells / P. I. Belobrov, A. A. Evstrapov, E. N. Esimbekova [et al.] // Journal of Physics: Conference Series : Institute of Physics Publishing, 2019. - Vol. 1379: Joint IMEKO TC1-TC7-TC13-TC18 Symposium 2019 (2 July 2019 through 5 July 2019, ) Conference code: 156337, Is. 1. - Ст. 012036, DOI 10.1088/1742-6596/1379/1/012036
Кл.слова (ненормированные):
Fluidic devices -- Industrial waste disposal -- Biological measurement -- Bioluminescent systems -- Continuous measurements -- Droplet-based microfluidics -- Industrial enterprise -- Microfluidic platforms -- Numerical variables -- Silicon photomultiplier -- Microfluidics
Аннотация: Here we discuss the concepts of "biological quantity" and "nominal property" within the framework of the problem of biological measurements based on new specific results of biological analysis using a microfluidic platform and chips developed by our team earlier. It was shown that based on different microfluidic platforms it is possible to develop chips with a polyenzymatic bioluminescent system NAD(P)H:FMN-oxidoreductase-luciferase (Red + Luc), which can be used in various areas of biological analysis. Thus, disposable microfluidic chips with Red + Luc system suitable for field and indoor use were developed using continuous flow microfluidic platform. The use of droplet-based microfluidic platform allowed to develop microfluidic chips with Red + Luc system for long-term continuous measurements of water samples, for example, in places of waste disposal by industrial enterprises. The reference for comparing different biological quantities with each other in the proposed system was a photodetector, which converted non-numeric values and nominal properties recorded by a biological module Red + Luc into numerical variables. Such a reference was implemented as a portable luminometer based on silicon photomultiplier. The results allow to perform other biological measurements and to start the discussion of modern biological concepts in the language of biological measures. © 2019 IOP Publishing Ltd. All rights reserved.

Scopus
Держатели документа:
Laboratory of Bioluminescent Biotechnology, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Institute for Analytical Instrumentation RAS, St.-Petersburg, 190103, Russian Federation
Laboratory of Photobiology, Institute of Biophysics SB RAS, Krasnoyarsk, 660036, Russian Federation
Laboratory for Digital Controlled Drugs and Theranostics, Federal Research Center krasnoyarsk Science Center SB RAS, Krasnoyarsk, 660036, Russian Federation
Research Institute of Molecular Medicine and Pathobiochemistry, Krasnoyarsk State Medical University Named after Prof. V.F. Voino-Yasenetsky, Krasnoyarsk, 660022, Russian Federation

Доп.точки доступа:
Belobrov, P. I.; Evstrapov, A. A.; Esimbekova, E. N.; Denisov, I. A.; Lukyanenko, K. A.; Osipova, E. D.; Yakimov, A. S.

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17.

Effect of viscosity on efficiency of enzyme catalysis of bacterial luciferase coupled with lactate dehydrogenase and NAD(P)H:FMN-Oxidoreductase / O. S. Sutormin [et al.] // Mol. Cat. - 2018. - Vol. 458. - P60-66, DOI 10.1016/j.mcat.2018.08.012 . - ISSN 2468-8231
Кл.слова (ненормированные):
Bioluminescence -- Coupling of enzymes -- In vivo simulated media -- Metabolic chain -- Protein stability
Аннотация: One of the current trends of the modern biology figures out cellular enzyme behaviour. Numerous researches look more closely at the chemical composition of creating in vivo simulated media conditions. The aim of this work was to find out a thermodynamic cooperativity of enzymes in a triple-enzyme chain (lactate dehydrogenase + NAD(P)H: FMN-oxidoreductase + bacterial luciferase) under in vivo simulated condition. The thermodynamic cooperativity effects were found out based on the influence of the viscogens (glycerol and sucrose) on the thermal stability of the triple-enzyme system. The results showed that the viscogens do not lead to an increase in the thermal stability of the triple-enzyme system. In addition, organic solvents (sucrose and glycerol) added as viscous agents to the reaction medium altered the kinetics of this triple-enzyme chain, including changing the light emission decay constant (kdec) and quantum yield of luminescence (Q). Plus, sucrose was found to be more efficient in limiting the flexibility of enzymes than glycerol. The high sensitivity of the triple-enzyme system to the viscogens may be connected with a fact that lactate dehydrogenase does not bound with couple enzyme system NAD(P)H: FMN-oxidoreductase + bacterial luciferase inside the real cell. Since this approach may be used as a method to understand the real connection between enzymes in cellular multi-enzyme metabolic chains inside the luminous bacteria cell. © 2018 Elsevier B.V.

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Держатели документа:
Department of Biophysics, Institute of Fundamental Biology and Biotechnology, Siberian Federal University, Krasnoyarsk, Russian Federation
Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Federal Research Center ‘Krasnoyarsk Science Center SB RAS’, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Sutormin, O. S.; Sukovataya, I. E.; Pande, S.; Kratasyuk, V. A.

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18.

Enzymatic bioassay of soil: Sensitivity comparison of mono-, double- And triple-enzyme systems to soil toxicants / O. S. Sutormin [и др.] // Tsitologiya. - 2018. - Vol. 60, Is. 10. - С. 826-829, DOI 10.7868/S0041377118100132 . - ISSN 0041-3771
Кл.слова (ненормированные):
Bacterial luciferase -- Bioluminescent analysis -- Coupled enzyme systems -- Ecological monitoring -- Enzymatic toxicity bioassays -- Lactate dehydrogenase -- NADH:FMN-oxidoreductase -- Soil
Аннотация: In this paper, we have investigated the possibilities of application of enzymatic systems with increasing chain length as a bioassay to evaluate the soil contamination status. The sensitivity of monoenzyme reaction as well as double- and triple-enzyme chains based on NAD(P)H:FMN-oxidoreductase and luciferase of luminous bacteria and lactate dehydrogenase to pesticides and copper ions in water and water extracts from soils were estimated. For this, the toxicological parameter IC 20 reflecting the sensitivity limit of the enzyme system to the to-xicant was used. It was revealed that elongation of the coupled enzyme chain (from mono- to triple-enzyme) increases the sensitivity of the bioassay, in some cases by several orders of magnitude. This pattern can be used as a tool to improve the properties of enzymic bioassays. The effect of extracts from uncontaminated soils of various types on enzymatic systems also differs, which makes possible to design the specialized enzymatic bioassays as well. © 2018 Sankt Peterburg.All rights reserved.

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Держатели документа:
Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics Siberian Branch of RAS, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Sutormin, O. S.; Kolosova, E. M.; Nemtseva, A. V.; Iskorneva, I. V.; Lisitsa, A. A.; Matvienko, V. S.; Esimbekova, A. N.; Kratasyuk, V. A.

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19.

Bioluminescent assay for toxicological assessment of nanomaterials / E. N. Esimbekova [et al.] // Dokl. Biochem. Biophys. - 2017. - Vol. 472, Is. 1. - P60-63, DOI 10.1134/S1607672917010173. - Cited References:15. - We are sincerely grateful to the staff of the Institute of Physiological Active Compounds (Kharkiv, Ukraine) for providing fullerene samples. This study was supported by the Russian Science Foundation (project no. 16-14-10115). . - ISSN 1607-6729. - ISSN 1608-3091

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
LUMINOUS BACTERIA
TOXICITY
Аннотация: A new method for assessing biotoxicity of nanomaterials, based on the use of soluble bioluminescent coupled enzyme system NAD(P)ai...H:FMN oxidoreductase and luciferase, is proposed. The results of this study indicate a significant adverse biological effect exerted by nanoparticles at the molecular level. It was found that the most toxic nanoparticles the nanoparticles are based on copper and copper oxide, as well as single-walled carbon nanotubes and multi-walled carbon nanofibers, which are referred to hazard class II.

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Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.
Krasnoyarsk State Agr Univ, Krasnoyarsk, Russia.

Доп.точки доступа:
Esimbekova, E. N.; Nemtseva, E. V.; Kirillova, M. A.; Asanova, A. A.; Kratasyuk, V. A.; Russian Science Foundation [16-14-10115]

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20.

Bioluminescent Enzymatic Assay as a Tool for Studying Antioxidant Activity and Toxicity of Bioactive Compounds / N. S. Kudryasheva [et al.] // Photochem. Photobiol. - 2017. - Vol. 93, Is. 2. - P536-540, DOI 10.1111/php.12639. - Cited References:40. - The work was supported by the Russian Foundation for Basic Research, Grants 15-03-06786 and 15-43-04377-sibir; the state budget allocated to the fundamental research at the Russian Academy of Sciences (project 01201351504). . - ISSN 0031-8655. - ISSN 1751-1097

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
LUMINOUS MARINE-BACTERIA
HUMIC SUBSTANCES
DETOXIFICATION PROCESSES
Аннотация: A bioluminescent assay based on a system of coupled enzymatic reactions catalyzed by bacterial luciferase and NADH:FMN-oxidoreductase was developed to monitor toxicity and antioxidant activity of bioactive compounds. The assay enables studying toxic effects at the level of biomolecules and physicochemical processes, as well as determining the toxicity of general and oxidative types. Toxic and detoxifying effects of bioactive compounds were studied. Fullerenols, perspective pharmaceutical agents, nanosized particles, water-soluble polyhydroxylated fullerene-60 derivatives were chosen as bioactive compounds. Two homologous fullerenols with different number and type of substituents, C60O2-4(OH)(20-24) and Fe0.5C60(OH) O-x(y) (x + y = 40-42), were used. They suppressed bioluminescent intensity at concentrations 0.01 g L-1 and 0.001 g L-1 for C60O2-4(OH)(20-24) and Fe0.5C60(OH)(x)O-y, respectively; hence, a lower toxicity of C60O2-4(OH)(20-24) was demonstrated. Antioxidant activity of fullerenols was studied in model solutions of organic and inorganic oxidizers; changes in toxicities of general and oxidative type were determined; detoxification coefficients were calculated. Fullerenol C60O2-4(OH)(20-24) revealed higher antioxidant ability at concentrations 10(-17)-10(-5) g L-1. The difference in the toxicity and antioxidant activity of fullerenols was explained through their electron donor/acceptor properties and different catalytic activity. Principles of bioluminescent enzyme assay application for evaluating the toxic effect and antioxidant activity of bioactive compounds were summarized and the procedure steps were described.

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Держатели документа:
Inst Biophys SB RAS, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.
Natl Res Tomsk Polytech Univ, Tomsk, Russia.
Inst Phys SB RAS, Krasnoyarsk, Russia.

Доп.точки доступа:
Kudryasheva, Nadezhda S.; Kovel, Ekaterina S.; Sachkova, Anna S.; Vorobeva, Anna A.; Isakova, Viktoriya G.; Churilov, Grigoriy N.; Russian Foundation for Basic Research [15-03-06786, 15-43-04377-sibir]; Russian Academy of Sciences [01201351504]

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