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Controlled expression of bacterial luminescence genes cloned in a multicopy recombinant plasmid [Text] / E. E. Maksimova [et al.] // Microbiology. - 1997. - Vol. 66, Is. 2. - P. 184-187. - Cited References: 17 . - ISSN 0026-2617

РУБ Microbiology
Рубрики:
GENETICALLY-MODIFIED MICROORGANISMS
BIOLUMINESCENCE
ENVIRONMENT
Кл.слова (ненормированные):
recombinant plasmid -- Escherichia coli -- luminescence -- catabolite repression
Аннотация: Luminescence and growth responses of the recombinant strain Escherichia coil Z905 (Ap(r)Lux(+)) to different concentrations of ampicillin depended on the source of carbon and energy. When glycerol was used as the substrate, the intensity of luminescence rose with the ampicillin concentration in the medium. Glucose caused catabolite repression of cell luminescence up to the late stationary phase, and ampicillin enhanced this effect. The feasibility of controlling the expression of cloned lux genes was shown.

WOS : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Maksimova, E.E.; Popova, L.Y.; Kargatova, T.V.; Shpagina, V.V.

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Fatty acid composition and polyhydroxyalkanoates production by Cupriavidus eutrophus B-10646 cells grown on different carbon sources [Text] / N. Zhila, G. Kalacheva, T. Volova // Process Biochem. - 2015. - Vol. 50, Is. 1. - P69-78, DOI 10.1016/j.procbio.2014.10.018. - Cited References:66. - This study was supported by the Russian Science Foundation (Grant No.14-26-00039). . - ISSN 1359-5113. - ISSN 1873-3298

РУБ Biochemistry & Molecular Biology + Biotechnology & Applied Microbiology
Рубрики:
HYDROGEN-OXIDIZING BACTERIA
CHAIN TRANSFER AGENTS
RALSTONIA-EUTROPHA
Кл.слова (ненормированные):
Cupriavidus eutrophus -- Fatty acid -- Polyhydroxyalkanoates -- Molecular -- weight -- Different substrates
Аннотация: Growth of Cupriavidus eutrophus B-10646, fatty acid (FA) composition of the lipids in the cytoplasmic membrane and cell wall, and polyhydroxyalkanoate (PHA) accumulation by the cells of this new promising producer of PHAs were studied in batch culture, using various carbon sources (fructose, glucose, oleic acid, sunflower seed oil, glycerol, CO2:O-2:H-2). In cells grown on sugars and in autotrophic culture, during the phase of active cell growth, the major FAs of the lipids of cytoplasmic membrane (CMLs) were palmitic, palmitoleic, and cis-vaccenic acids. In the stationary phase of cultivation, cyclopropane FAs were also detected. When carbohydrate substrate was replaced by oleic acid or sunflower seed oil, the proportion of oleic acid in the total FAs of the cytoplasmic membrane increased considerably. In addition to that, the lipid FAs of bacterial cells grown on sunflower seed oil also contained linoleic acid, which is the major acid of sunflower seed oil. The major acids of strongly bound lipids of cell wall (SBLs) were myristic acid and long-chain-length beta-hydroxy acids. Molecular weight of the polymer was measured during its accumulation in cells; for the first time, the presence of 2 polymer fractions (high-molecular weight and low-molecular-weight ones) was recorded in cells with a low polymer content. (C) 2014 Elsevier Ltd. All rights reserved.

WOS
Держатели документа:
SB RAS, Inst Biophys, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.
ИБФ СО РАН

Доп.точки доступа:
Zhila, Natalia; Kalacheva, Galina; Volova, Tatiana; Russian Science Foundation [14-26-00039]

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Production and properties of bacterial cellulose by the strain Komagataeibacter xylinus B-12068 / T. G. Volova [et al.] // Appl. Microbiol. Biotechnol. - 2018. - P1-12, DOI 10.1007/s00253-018-9198-8 . - ISSN 0175-7598
Кл.слова (ненормированные):
Bacterial cellulose -- Growth conditions -- Komagataeibacter xylinus -- Biocompatibility -- Cell culture -- Cellulose -- Cultivation -- Glucose -- 3t3 mouse fibroblasts -- Bacterial cellulose -- Cultivation conditions -- Emission spectrometry -- Ethanol concentrations -- Growth conditions -- Komagataeibacter xylinus -- Physical and mechanical properties -- Substrates
Аннотация: A strain of acetic acid bacteria, Komagataeibacter xylinus B-12068, was studied as a source for bacterial cellulose (BC) production. The effects of cultivation conditions (carbon sources, temperature, and pH) on BC production and properties were studied in surface and submerged cultures. Glucose was found to be the best substrate for BC production among the sugars tested; ethanol concentration of 3% (w/v) enhanced the productivity of BC. Optimization of medium and cultivation conditions ensures a high production of BC on glucose and glycerol, up to 2.4 and 3.3 g/L/day, respectively. C/N elemental analysis, emission spectrometry, SEM, DTA, and X-ray were used to investigate the structure and physical and mechanical properties of the BC produced under different conditions. MTT assay and SEM showed that native cellulose membrane did not cause cytotoxicity upon direct contact with NIH 3T3 mouse fibroblast cells and was highly biocompatible. © 2018 Springer-Verlag GmbH Germany, part of Springer Nature

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

Доп.точки доступа:
Volova, T. G.; Prudnikova, S. V.; Sukovatyi, A. G.; Shishatskaya, E. I.

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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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Polyhydroxyalkanoate synthesis based on glycerol and implementation of the process under conditions of pilot production / T. Volova [et al.] // Appl. Microbiol. Biotechnol. - 2018, DOI 10.1007/s00253-018-9460-0 . - Article in press. - ISSN 0175-7598
Кл.слова (ненормированные):
Glycerol -- Polyhydroxyalkanoates -- Productivity -- Properties -- Synthesis -- Batch cell culture -- Fermentation -- Organic polymers -- Pilot plants -- Productivity -- Purification -- Synthesis (chemical) -- Degree of crystallinity -- Fed-batch fermentation -- Poly-3-hydroxybutyrate -- Poly-hydroxyalkanoate -- Polyhydroxyalkanoates -- Properties -- Temperature characteristic -- Volumetric productivity -- Glycerol
Аннотация: The present study addresses the synthesis and properties of polyhydroxyalkanoates (PHA) of different composition synthesized by Cupriavidus eutrophus B-10646 using glycerol as a carbon substrate. Poly(3-hydroxybutyrate) [P(3HB)] was effectively synthesized in fed-batch culture in a 30-L fermenter on glycerol of various purification degrees, with 99.5, 99.7, and 82.1% content of the main component. Purified glycerol (99.7%) was used for 150-L pilot scale fermentation. The total biomass and P(3HB) concentration reached 110 and 85.8 g/L, respectively, after 45 h of fed-batch fermentation. An average volumetric productivity of P(3HB) was 1.83 g/(L h). The degree of crystallinity and molecular weight of P(3HB) synthesized on glycerol were lower than and temperature characteristics were the same as those of P(3HB) synthesized on sugars. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.

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Держатели документа:
Siberian Federal University, 79 Svobodnyi Ave, Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Akademgorodok, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Volova, T.; Demidenko, A.; Kiselev, E.; Baranovskiy, S.; Shishatskaya, E.; Zhila, N.

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Synthesis of Polyhydroxyalkanoates by Hydrogen-Oxidizing Bacteria in a Pilot Production Process / T. Volova [et al.] // Biomacromolecules. - 2019, DOI 10.1021/acs.biomac.9b00295 . - Article in press. - ISSN 1525-7797
Кл.слова (ненормированные):
Crystallinity -- Fructose -- Glucose -- Pilot plants -- Purification -- Substrates -- Terpolymers -- Carbon substrates -- Cell concentrations -- Degree of crystallinity -- Hydrogen-oxidizing bacteria -- Polies (3 hydroxybutyrate co 3 hydroxyvalerate co 4 hydroxybutyrate) -- Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) -- Polyhydroxyalkanoates -- Yield coefficient -- Glycerol
Аннотация: The synthesis of polyhydroxyalkanoates (PHAs) was scaled up to pilot production in a 150-L fermenter on sugars (fructose and glucose) and purified and crude glycerol in a culture of the wild-type strain Cupriavidus necator B-10646. Over 60 h of cultivation, a cell concentration of 150-160 g/L was obtained on purified glycerol and glucose; cultivation on fructose and crude glycerol resulted in a cell concentration of 130 ± 10 g/L. Polymer content and yield coefficients for the biomass were similar on all substrates (80-85 wt % and 0.29-0.33 kg biomass/kg carbon substrate, respectively). Copolymers poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) and terpolymers poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) having a decreased degree of crystallinity (36-46%) were first synthesized in the scaled-up process using C. necator B-10646 cultivated on glycerol. These results will provide the basis for scaling-up PHA synthesis in an organotrophic C. necator B-10646 culture. © 2019 American Chemical Society.

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Держатели документа:
Siberian Federal University, 79 Svobodnyi Av., Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, Federal Research Center Krasnoyarsk Science Center SB RAS, 50/50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Volova, T.; Kiselev, E.; Zhila, N.; Shishatskaya, E.

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Viscous Media Slow Down the Decay of the Key Intermediate in Bacterial Bioluminescent Reaction / A. E. Lisitsa, L. A. Sukovatyi, V. A. Kratasyuk, E. V. Nemtseva // Doklad. Biochem. Biophys. - 2020. - Vol. 492, Is. 1. - P162-165, DOI 10.1134/S1607672920020106 . - ISSN 1607-6729
Кл.слова (ненормированные):
bacterial luciferase -- bioluminescence -- diffusional restriction -- enzyme reaction intermediate -- molecular dynamics method -- stopped-flow technique -- viscous microenvironment
Аннотация: Abstract: The effects of medium viscosity on the decay rate of the 4a-hydroperoxyflavin intermediate of the bioluminescent reaction was investigated. It was found that at low concentrations of glycerol or sucrose (viscosity 1.1–1.3 cP) the decay rate rises, whereas a further increase in viscosity to 6.2 cP leads to a decrease in the decay rate following a power function with an exponent of 0.82–0.84. Using molecular dynamics methods, it was shown that the presence of glycerol and sucrose molecules causes a change in the mobility of the amino acid residues in the active center of luciferase, particularly those responsible for binding of flavin. The results obtained are indicative of two opposite effects of viscous media with glycerol and sucrose: (1) destabilization of 4a-hydroperoxyflavin due to a change in the structural and dynamic properties of the protein and (2) stabilization of this intermediate by the decrease in the diffusion rate of its decay products. © 2020, Pleiades Publishing, Ltd.

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

Доп.точки доступа:
Lisitsa, A. E.; Sukovatyi, L. A.; Kratasyuk, V. A.; Nemtseva, E. V.

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Viscous Media Slow Down the Decay of the Key Intermediate in Bacterial Bioluminescent Reaction / A. E. Lisitsa, L. A. Sukovatyi, V. A. Kratasyuk, E. V. Nemtseva // Dokl. Biochem. Biophys. - 2020. - Vol. 492, Is. 1. - P162-165, DOI 10.1134/S1607672920020106. - Cited References:15. - This work was supported by the Ministry of Science and Higher Education of the Russian Federation (project nos. 6.7734.2017 and 01201351504). . - ISSN 1607-6729. - ISSN 1608-3091

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
LUCIFERASE
Кл.слова (ненормированные):
bacterial luciferase -- bioluminescence -- viscous microenvironment -- stopped-flow technique -- molecular dynamics method -- enzyme reaction -- intermediate -- diffusional restriction
Аннотация: The effects of medium viscosity on the decay rate of the 4a-hydroperoxyflavin intermediate of the bioluminescent reaction was investigated. It was found that at low concentrations of glycerol or sucrose (viscosity 1.1-1.3 cP) the decay rate rises, whereas a further increase in viscosity to 6.2 cP leads to a decrease in the decay rate following a power function with an exponent of 0.82-0.84. Using molecular dynamics methods, it was shown that the presence of glycerol and sucrose molecules causes a change in the mobility of the amino acid residues in the active center of luciferase, particularly those responsible for binding of flavin. The results obtained are indicative of two opposite effects of viscous media with glycerol and sucrose: (1) destabilization of 4a-hydroperoxyflavin due to a change in the structural and dynamic properties of the protein and (2) stabilization of this intermediate by the decrease in the diffusion rate of its decay products.

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

Доп.точки доступа:
Lisitsa, A. E.; Sukovatyi, L. A.; Kratasyuk, V. A.; Nemtseva, E. V.; Ministry of Science and Higher Education of the Russian Federation [6.7734.2017, 01201351504]

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Polymer Films of Poly-3-hydroxybutyrate Synthesized byCupriavidus necatorfrom Different Carbon Sources / E. Shishatskaya, I. Nemtsev, A. Lukyanenko [et al.] // J. Polym. Environ. - 2020, DOI 10.1007/s10924-020-01924-3. - Cited References:54. - The reported study was funded by RFBR and KKRF Grant No. 19-43-240012 "Biological and physical principles of production of new generation biomaterials". The work was carried out as part of the State Assignment of the Ministry of Education and Science of the Russian Federation No. FSRZ-2020-0006. The authors would like to express their special thanks to Krasnoyarsk Regional Center of Research Equipment of Federal Research Center "Krasnoyarsk Science Center SB RAS" for providing equipment to ensure the accomplishment of this project. . - Article in press. - ISSN 1566-2543. - ISSN 1572-8919

РУБ Engineering, Environmental + Polymer Science
Рубрики:
CHEMOMECHANICAL PROPERTIES
RALSTONIA-EUTROPHA
SURFACE-ROUGHNESS
Кл.слова (ненормированные):
Degradable P(3HB) -- Various carbon substrates -- Films -- Structure -- Properties
Аннотация: Films were prepared from 2% solutions of biodegradable poly-3-hydroxybutyrate [P(3HB)] and investigated. The polymer was synthesized by theCupriavidus necatorB-10646 bacterium cultivated using various carbon sources (glucose and glycerol of different degrees of purity, containing 0.3 to 17.93% impurities). Glycerol as the substrate influenced molecular-weight properties and crystallinity of the polymer without affecting its temperature characteristics. The P(3HB) specimens synthesized from glycerol had reduced M-w(300-400 kDa) and degree of crystallinity (50-55%) compared to the specimens synthesized from glucose (860 kDa and 76%, respectively). The low-crystallinity P(3HB) specimens, regardless of the degree of purity of glycerol, produced a beneficial effect on the properties of polymer films, which had a better developed folded surface and increased hydrophilicity. The values of the highest roughness (R-a) of the films synthesized from glycerol were 1.8 to 4.0 times lower and the water angles 1.4-1.6 times smaller compared to the films synthesized from glucose (71.75 nm and 87.4 degrees, respectively). Those films performed better as cell scaffolds: the number of viable NIH fibroblasts was 1.7-1.9 times higher than on polystyrene (control) or films of P(3HB) synthesized from glucose.

WOS
Держатели документа:
Siberian Fed Univ, 79 Svobodnyi Av, Krasnoyarsk 660041, Russia.
RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, Inst Biophys, Krasnoyarsk, Russia.
RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, LV Kirenskii Inst Phys, Krasnoyarsk, Russia.

Доп.точки доступа:
Shishatskaya, Ekaterina; Nemtsev, Ivan; Lukyanenko, Anna; Vasiliev, Alexander; Kiselev, Evgeniy; Sukovatyi, Aleksey; Volova, Tatiana; RFBRRussian Foundation for Basic Research (RFBR); KKRF [19-43-240012]; Ministry of Education and Science of the Russian FederationMinistry of Education and Science, Russian Federation [FSRZ-2020-0006]

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10.

Polymer Films of Poly-3-hydroxybutyrate Synthesized by Cupriavidus necator from Different Carbon Sources / E. Shishatskaya, I. Nemtsev, A. Lukyanenko [et al.] // J. Polym. Environ. - 2021. - Vol. 29, Is. 3. - P837-850, DOI 10.1007/s10924-020-01924-3 . - ISSN 1566-2543
Кл.слова (ненормированные):
Degradable P(3HB) -- Films -- NIH 3T3 fibroblasts -- Properties -- Structure -- Various carbon substrates -- Carbon -- Carbon films -- Cell culture -- Chlorine containing polymers -- Crystallinity -- Glucose -- Glycerol -- Scaffolds (biology) -- Semiconducting films -- Beneficial effects -- Cell scaffold -- Degree of crystallinity -- Different carbon sources -- Low crystallinity -- Poly-3-hydroxybutyrate -- Temperature characteristic -- Weight Properties -- Polymer films -- Bacteria (microorganisms) -- bacterium B -- Cupriavidus necator
Аннотация: Films were prepared from 2% solutions of biodegradable poly-3-hydroxybutyrate [P(3HB)] and investigated. The polymer was synthesized by the Cupriavidus necator B-10646 bacterium cultivated using various carbon sources (glucose and glycerol of different degrees of purity, containing 0.3 to 17.93% impurities). Glycerol as the substrate influenced molecular-weight properties and crystallinity of the polymer without affecting its temperature characteristics. The P(3HB) specimens synthesized from glycerol had reduced Mw (300–400 kDa) and degree of crystallinity (50–55%) compared to the specimens synthesized from glucose (860 kDa and 76%, respectively). The low-crystallinity P(3HB) specimens, regardless of the degree of purity of glycerol, produced a beneficial effect on the properties of polymer films, which had a better developed folded surface and increased hydrophilicity. The values of the highest roughness (Ra) of the films synthesized from glycerol were 1.8 to 4.0 times lower and the water angles 1.4–1.6 times smaller compared to the films synthesized from glucose (71.75 nm and 87.4°, respectively). Those films performed better as cell scaffolds: the number of viable NIH fibroblasts was 1.7–1.9 times higher than on polystyrene (control) or films of P(3HB) synthesized from glucose. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.

Scopus
Держатели документа:
Siberian Federal University, 79 Svobodnyi Av., Krasnoyarsk, 660041, Russian Federation
Institute of Biophysics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, Russian Federation
L.V. Kirenskii Institute of Physics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Shishatskaya, E.; Nemtsev, I.; Lukyanenko, A.; Vasiliev, A.; Kiselev, E.; Sukovatyi, A.; Volova, T.

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11.

The Effect of Osmolytes on the Bioluminescent Reaction of Bacteria: Structural and Dynamic Properties / L. A. Sukovatyi, A. E. Lisitsa, V. A. Kratasyuk, E. V. Nemtseva // Biophysics. - 2020. - Vol. 65, Is. 6. - P966-971, DOI 10.1134/S0006350920060202 . - ISSN 0006-3509
Кл.слова (ненормированные):
bacterial luciferase -- bioluminescence -- luminous bacteria -- molecular dynamic -- osmolyte -- protein structure and dynamics
Аннотация: The effects of viscous media with glycerol and sucrose (10–40%) on the kinetics of the bacterial bioluminescent reaction have been investigated by stopped-flow technique. Increment of quantum yield in media with 10% of both osmolytes was shown. Higher concentrations of glycerol, up to 30–40%, were found to reduce the efficiency of the reaction, while this effect was not observed in the media with sucrose. The molecular dynamics simulation was used to study the structure of bacterial luciferase surrounded by either water molecules solely or by mixture of water with various numbers of glycerol/sucrose molecules. It was found that both cosolvents at studied concentrations did not cause a significant change in conformation of bacterial luciferase. The calculated root-mean-square fluctuation for C?-atoms of bacterial luciferase ?-subunit indicated that the higher flexibility of the enzyme mobile loop could be responsible for increment of quantum yield in the presence of 10% of both osmolytes. The active site of bacterial luciferase was found to be accessible for glycerol molecules while sucrose did not enter catalytic gorge. Moreover, at 30 and 40% concentration the glycerol molecules were found to locate in the active site of bacterial luciferase throughout the whole simulation time. © 2020, Pleiades Publishing, Inc.

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

Доп.точки доступа:
Sukovatyi, L. A.; Lisitsa, A. E.; Kratasyuk, V. A.; Nemtseva, E. V.

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12.

Biodegradable polymers - Perspectives and applications in agriculture / E. G. Kiselev, N. O. Zhila, T. G. Volova // IOP Conference Series: Earth and Environmental Science : IOP Publishing Ltd, 2021. - Vol. 689: 2020 International Conference on Germany and Russia: Ecosystems Without Borders, EcoSystConfKlgtu 2020 (5 October 2020 through 10 October 2020, ) Conference code: 167944, Is. 1. - Ст. 012036, DOI 10.1088/1755-1315/689/1/012036
Кл.слова (ненормированные):
Biodegradable polymers -- Ecosystems -- Fungi -- Glycerol -- Monounsaturated fatty acids -- Oilseeds -- Pesticides -- Substrates -- Sunflower oil -- Fenoxaprop-p-ethyl -- Natural materials -- Pesticide formulations -- Poly-3-hydroxybutyrate -- Polyhydroxyalkanoates -- Productive process -- Strategy of constructions -- Various substrates -- Palm oil
Аннотация: The paper presents a brief overview of the results of the implementation of the project "Agropreparations of the new generation: a strategy of construction and realization". The first part contains the analysis of the growth of the wild-type strain Cupriavidus necator B-10646 (formerly eutrophus) and the synthesis of polyhydroxyalkanoates by this strain on various substrates: glycerol, palm oil, Siberian oil seed, sunflower seed oils, and oleic acid. On refined glycerin, a highly productive process is implemented when scaling up, allowing to obtain 128 ± 11 g / L PHA. Evaluation of oils has shown that palm oil is the best carbon substrate. The second part presents the results of the development of environmentally friendly slow-release pesticide formulations. They are a degradable matrix of poly-3-hydroxybutyrate mixed with natural materials (peat, clay, wood flour), into which a pesticide (metribuzin, tribenuron-methyl, fenoxaprop-P-ethyl, azoxystrobin, epoxiconazole, and tebuconazole) has been. The developed preparations showed high activity against pathogenic fungi and weeds and had a much weaker negative effect on the soil microflora. Studies of the degradation of the developed preparations and the release of pesticides into the soil confirm their effectiveness over a long period of time, up to 90 days. © Published under licence by IOP Publishing Ltd.

Scopus
Держатели документа:
School of Fundamental Biology and Biotechnology, Siberian Federal University, Krasnoyarsk, Russian Federation
Laboratory of Chemoautotrophic Biosynthesis, Institute of Biophysics, SB, RAS, Federal Research Center, Krasnoyarsk Science Center SB RAS, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Kiselev, E. G.; Zhila, N. O.; Volova, T. G.

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13.

Development of the method to produce functionally active recombinant streptavidin in escherichia coli cells / E. E. Bashmakova, A. N. Kudryavtsev, L. A. Frank // J. Sib. Fed. Univ. - Biol. - 2020. - Vol. 13, Is. 2. - С. 218-229, DOI 10.17516/1997-1389-0324 . - ISSN 1997-1389
Кл.слова (ненормированные):
E. coli protein-producing strain -- Microanalysis -- Recombinant streptavidin
Аннотация: Streptavidin is a homotetrameric protein produced by Streptomyces avidinii, each subunit of which binds biotin (vitamin H), forming a stable complex (Kd = 10-15 M). Streptavidin-biotin coreaction is widely used in analytical systems, for targeted delivery of compounds, for affinity purification, etc. The aim of this study was to develop a rational technique to produce functionally active recombinant streptavidin. Recombinant Escherichia coli strains producing minimal core and full-sized streptavidin variants were obtained. The E. coli BL21 Codon Plus (DE3) RIPL, as host cells, and the pET19b plasmid carrying gene of minimally-sized core (miniSAV) or full-sized (SAV) streptavidin were used. Synthesis of miniSAV results in its localization as insoluble inclusion bodies. Denatured miniSAV yield was 130 mg per liter of E. coli c ulture. T he r enaturation g ives o nly 10- 15 % of the functionally active protein. Full-sized streptavidin localizes in the cytoplasm in a soluble state, but its toxicity causes low yield of the protein (10-13 mg per liter of the culture). The induction of SAV synthesis at the end of the logarithmic stage of cell growth was found to increase the yield of SAV approximately 2-fold. The yield of functionally active protein was 30 mg per liter culture. SAV was produced practically in individual state after affine chromatography on 2-iminobiotin agarose. One molecule of full-sized streptavidin bound 3.9 biotin molecules as was shown by colorimetric analysis using HABA (4-hydroxyazobenzene-2-carboxylic acid). Both streptavidins form sandwichtype complexes with biotinylated molecules in solid-phase microassay conditions. E. coli BL21 Codon Plus (DE3) RIPL/pET19bSAV strain was stable during storage with 20 % glycerol at -70 °C, which was shown by repeated two-year reseeding. The streptavidin producing strain (E. coli BL21 Codon Plus (DE3) RIPL/pET19bSAV) is deposited in the Collection for extremophile microorganisms and type cultures (Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk), No. 3505. The method for producing functionally active recombinant streptavidin developed in this study ensures its availability for biotechnological research. © Siberian Federal University. All rights reserved.

Scopus
Держатели документа:
Institute of Biophysics SB RAS, FRC Krasnoyarsk Science Center SB RAS, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Bashmakova, E. E.; Kudryavtsev, A. N.; Frank, L. A.

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14.

Mechanisms of viscous media effects on elementary steps of bacterial bioluminescent reaction / A. E. Lisitsa, L. A. Sukovatyi, S. I. Bartsev [et al.] // Int. J. Mol. Sci. - 2021. - Vol. 22, Is. 16. - Ст. 8827, DOI 10.3390/ijms22168827 . - ISSN 1661-6596
Кл.слова (ненормированные):
Bacterial luciferase -- Diffusion limitation -- Non-steady-state reaction kinetics -- Viscosity
Аннотация: Enzymes activity in a cell is determined by many factors, among which viscosity of the microenvironment plays a significant role. Various cosolvents can imitate intracellular conditions in vitro, allowing to reduce a combination of different regulatory effects. The aim of the study was to analyze the media viscosity effects on the rate constants of the separate stages of the bacterial biolumi-nescent reaction. Non-steady-state reaction kinetics in glycerol and sucrose solutions was measured by stopped-flow technique and analyzed with a mathematical model developed in accordance with the sequence of reaction stages. Molecular dynamics methods were applied to reveal the effects of cosolvents on luciferase structure. We observed both in glycerol and in sucrose media that the stages of luciferase binding with flavin and aldehyde, in contrast to oxygen, are diffusion-limited. More-over, unlike glycerol, sucrose solutions enhanced the rate of an electronically excited intermediate formation. The MD simulations showed that, in comparison with sucrose, glycerol molecules could penetrate the active-site gorge, but sucrose solutions caused a conformational change of functionally important ?Glu175 of luciferase. Therefore, both cosolvents induce diffusion limitation of substrates binding. However, in sucrose media, increasing enzyme catalytic constant neutralizes viscosity effects. The activating effect of sucrose can be attributed to its exclusion from the catalytic gorge of luciferase and promotion of the formation of the active site structure favorable for the catalysis. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

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

Доп.точки доступа:
Lisitsa, A. E.; Sukovatyi, L. A.; Bartsev, S. I.; Deeva, A. A.; Kratasyuk, V. A.; Nemtseva, E. V.

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15.

Mechanisms of Viscous Media Effects on Elementary Steps of Bacterial Bioluminescent Reaction / A. E. Lisitsa, L. A. Sukovatyi, S. I. Bartsev [et al.] // Int. J. Mol. Sci. - 2021. - Vol. 22, Is. 16. - Ст. 8827, DOI 10.3390/ijms22168827. - Cited References:59. - The research was funded by the Ministry of Science and Higher Education of the Russian Federation (projects No. FSRZ-2020-0006); by RFBR, Krasnoyarsk Territory and Krasnoyarsk Regional Fund of Science (project No. 20-44-243002); by RFBR according to the research project No. 20-34-90118. . - ISSN 1422-0067

РУБ Biochemistry & Molecular Biology + Chemistry, Multidisciplinary
Рубрики:
FLAVIN INTERMEDIATE
   REDUCED FLAVIN
   RATE CONSTANTS
   LUCIFERASE
Кл.слова (ненормированные):
bacterial luciferase -- non-steady-state reaction kinetics -- viscosity -- diffusion limitation
Аннотация: Enzymes activity in a cell is determined by many factors, among which viscosity of the microenvironment plays a significant role. Various cosolvents can imitate intracellular conditions in vitro, allowing to reduce a combination of different regulatory effects. The aim of the study was to analyze the media viscosity effects on the rate constants of the separate stages of the bacterial bioluminescent reaction. Non-steady-state reaction kinetics in glycerol and sucrose solutions was measured by stopped-flow technique and analyzed with a mathematical model developed in accordance with the sequence of reaction stages. Molecular dynamics methods were applied to reveal the effects of cosolvents on luciferase structure. We observed both in glycerol and in sucrose media that the stages of luciferase binding with flavin and aldehyde, in contrast to oxygen, are diffusion-limited. Moreover, unlike glycerol, sucrose solutions enhanced the rate of an electronically excited intermediate formation. The MD simulations showed that, in comparison with sucrose, glycerol molecules could penetrate the active-site gorge, but sucrose solutions caused a conformational change of functionally important alpha Glu175 of luciferase. Therefore, both cosolvents induce diffusion limitation of substrates binding. However, in sucrose media, increasing enzyme catalytic constant neutralizes viscosity effects. The activating effect of sucrose can be attributed to its exclusion from the catalytic gorge of luciferase and promotion of the formation of the active site structure favorable for the catalysis.

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

Доп.точки доступа:
Lisitsa, Albert E.; Sukovatyi, Lev A.; Bartsev, Sergey, I; Deeva, Anna A.; Kratasyuk, Valentina A.; Nemtseva, Elena, V; Nemtseva, Elena; Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]; RFBR, Krasnoyarsk Territory and Krasnoyarsk Regional Fund of Science [20-44-243002]; RFBRRussian Foundation for Basic Research (RFBR) [20-34-90118]

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16.

Properties of degradable polyhydroxyalkanoates (Phas) synthesized by a new strain, cupriavidus necator ibp/sfu-1, from various carbon sources / N. O. Zhila, K. Yu. Sapozhnikova, E. G. Kiselev [et al.] // Polym. - 2021. - Vol. 13, Is. 18. - Ст. 3142, DOI 10.3390/polym13183142 . - ISSN 2073-4360
Кл.слова (ненормированные):
Cell growth and PHA synthesis -- Cupriavidus necator IBP/SFU-1 -- PHA composition and properties -- Polymer films -- Various carbon sources -- Biodegradable polymers -- Carbon -- Carbon films -- Cell proliferation -- Crystallinity -- Fructose -- Glucose -- Long Term Evolution (LTE) -- Oleic acid -- Organic carbon -- Palm oil -- Polydispersity -- Semiconducting films -- Autotrophics -- Carbon source -- Cell growth and PHA synthesis -- Cupriavidu necator IBP/SFU-1 -- PHA composition and property -- Plant oil -- Polyhydroxyalkanoates -- Property -- Synthesised -- Various carbon source -- Polymer films
Аннотация: The bacterial strain isolated from soil was identified as Cupriavidus necator IBP/SFU-1 and investigated as a PHA producer. The strain was found to be able to grow and synthesize PHAs under autotrophic conditions and showed a broad organotrophic potential towards different carbon sources: sugars, glycerol, fatty acids, and plant oils. The highest cell concentrations (7–8 g/L) and PHA contents were produced from oleic acid (78%), fructose, glucose, and palm oil (over 80%). The type of the carbon source influenced the PHA chemical composition and properties: when grown on oleic acid, the strain synthesized the P(3HB-co-3HV) copolymer; on plant oils, the P(3HB-co-3HV-co-3HHx) terpolymer, and on the other substrates, the P(3HB) homopolymer. The type of the carbon source influenced molecular-weight properties of PHAs: P(3HB) synthesized under autotrophic growth conditions, from CO2, had the highest number-average (290 ± 15 kDa) and weight-average (850 ± 25 kDa) molecular weights and the lowest polydispersity (2.9 ± 0.2); polymers synthesized from organic carbon sources showed increased polydispersity and reduced molecular weight. The carbon source was not found to affect the degree of crystallinity and thermal properties of the PHAs. The type of the carbon source determined not only PHA composition and molecular weight but also surface microstructure and porosity of the polymer films. The new strain can be recommended as a promising P(3HB) producer from palm oil, oleic acid, and sugars (fructose and glucose) and as a producer of P(3HB-co-3HV) from oleic acid and P(3HB-co-3HV-co-3HHx) from palm oil. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Scopus
Держатели документа:
Basic Department of Biotechnology, School of Fundamental Biology and Biotechnology, Siberian Federal University, 79 Svobodnyi Av., Krasnoyarsk, 660041, Russian Federation
Federal Research Center, “Krasnoyarsk Science Center SB RAS”, Institute of Biophysics SB RAS, 50/50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation
Federal Research Center, “Krasnoyarsk Science Center SB RAS”, L.V. Kirensky Institute of Physics SB RAS, 50/38 Akademgorodok, Krasnoyarsk, 660036, Russian Federation
Federal Research Center, “Krasnoyarsk Science Center of the Siberian Branch of the Russian Academy of Sciences”, 50 Akademgorodok, Krasnoyarsk, 660036, Russian Federation

Доп.точки доступа:
Zhila, N. O.; Sapozhnikova, K. Yu.; Kiselev, E. G.; Vasiliev, A. D.; Nemtsev, I. V.; Shishatskaya, E. I.; Volova, T. G.

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17.

Properties of Degradable Polyhydroxyalkanoates (PHAs) Synthesized by a New Strain, Cupriavidus necator IBP/SFU-1, from Various Carbon Sources / N. O. Zhila, K. Y. Sapozhnikova, E. G. Kiselev [et al.] // Polymers. - 2021. - Vol. 13, Is. 18. - Ст. 3142, DOI 10.3390/polym13183142. - Cited References:78. - This work was financially supported by Project "Agropreparations of the new generation: a strategy of construction and realization" (Agreement No 075-15-2021-626) in accordance with Resolution No 220 of the Government of the Russian Federation of 9 April 2010, "On measures designed to attract leading scientists to the Russian institutions of higher learning" (strain isolation, polymer synthesis and investigation), and by the State Assignment of the Ministry of Science and Higher Education of the Russian Federation No. FSRZ-2020-0006 (study of film properties). . - ISSN 2073-4360

РУБ Polymer Science
Рубрики:
RALSTONIA-EUTROPHA
   POLY(3-HYDROXYBUTYRATE) PRODUCTION
   PLANT OIL
   ACID
Кл.слова (ненормированные):
Cupriavidus necator IBP -- SFU-1 -- cell growth and PHA synthesis -- various -- carbon sources -- PHA composition and properties -- polymer films
Аннотация: The bacterial strain isolated from soil was identified as Cupriavidus necator IBP/SFU-1 and investigated as a PHA producer. The strain was found to be able to grow and synthesize PHAs under autotrophic conditions and showed a broad organotrophic potential towards different carbon sources: sugars, glycerol, fatty acids, and plant oils. The highest cell concentrations (7-8 g/L) and PHA contents were produced from oleic acid (78%), fructose, glucose, and palm oil (over 80%). The type of the carbon source influenced the PHA chemical composition and properties: when grown on oleic acid, the strain synthesized the P(3HB-co-3HV) copolymer; on plant oils, the P(3HB-co-3HV-co-3HHx) terpolymer, and on the other substrates, the P(3HB) homopolymer. The type of the carbon source influenced molecular-weight properties of PHAs: P(3HB) synthesized under autotrophic growth conditions, from CO2, had the highest number-average (290 +/- 15 kDa) and weight-average (850 +/- 25 kDa) molecular weights and the lowest polydispersity (2.9 +/- 0.2); polymers synthesized from organic carbon sources showed increased polydispersity and reduced molecular weight. The carbon source was not found to affect the degree of crystallinity and thermal properties of the PHAs. The type of the carbon source determined not only PHA composition and molecular weight but also surface microstructure and porosity of the polymer films. The new strain can be recommended as a promising P(3HB) producer from palm oil, oleic acid, and sugars (fructose and glucose) and as a producer of P(3HB-co-3HV) from oleic acid and P(3HB-co-3HV-co-3HHx) from palm oil.

WOS
Держатели документа:
Siberian Fed Univ, Basic Dept Biotechnol, Sch Fundamental Biol & Biotechnol, 79 Svobodnyi Av, Krasnoyarsk 660041, Russia.
SB RAS, Inst Biophys, Fed Res Ctr, Krasnoyarsk Sci Ctr, 50-50 Akademgorodok, Krasnoyarsk 660036, Russia.
SB RAS, LV Kirensky Inst Phys, Krasnoyarsk Sci Ctr, Fed Res Ctr, 50-38 Akademgorodok, Krasnoyarsk 660036, Russia.
Russian Acad Sci, Fed Res Ctr, Krasnoyarsk Sci Ctr, Siberian Branch, 50 Akademgorodok, Krasnoyarsk 660036, Russia.

Доп.точки доступа:
Zhila, Natalia O.; Sapozhnikova, Kristina Yu; Kiselev, Evgeniy G.; Vasiliev, Alexander D.; Nemtsev, Ivan, V; Shishatskaya, Ekaterina, I; Volova, Tatiana G.; Russian FederationRussian Federation [075-15-2021-626, 220]; State Assignment of the Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]

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18.

Bacterial Cellulose (BC) and BC Composites: Production and Properties / TGG Volova, SVV Prudnikova, EGG Kiselev [et al.] // Nanomaterials. - 2022. - Vol. 12, Is. 2. - Ст. 192, DOI 10.3390/nano12020192. - Cited References:113. - This research was financially supported by the State Assignment of the Ministry of Science and Higher Education of the Russian Federation No. FSRZ-2020-0006. . - ISSN 2079-4991

РУБ Chemistry, Multidisciplinary + Nanoscience & Nanotechnology + Materials
Рубрики:
SILVER NANOPARTICLES
GLUCONACETOBACTER-HANSENII
MICROBIAL CELLULOSE
Кл.слова (ненормированные):
bacterial cellulose -- composites -- production -- properties
Аннотация: The synthesis of bacterial cellulose (BC) by Komagataeibacter xylinus strain B-12068 was investigated on various C-substrates, under submerged conditions with stirring and in static surface cultures. We implemented the synthesis of BC on glycerol, glucose, beet molasses, sprat oil, and a mixture of glucose with sunflower oil. The most productive process was obtained during the production of inoculum in submerged culture and subsequent growth of large BC films (up to 0.2 m(2) and more) in a static surface culture. The highest productivity of the BC synthesis process was obtained with the growth of bacteria on molasses and glycerol, 1.20 and 1.45 g/L per day, respectively. We obtained BC composites with silver nanoparticles (BC/AgNPs) and antibacterial drugs (chlorhexidine, baneocin, cefotaxime, and doripenem), and investigated the structure, physicochemical, and mechanical properties of composites. The disc-diffusion method showed pronounced antibacterial activity of BC composites against E. coli ATCC 25922 and S. aureus ATCC 25923.

WOS
Держатели документа:
Siberian Fed Univ, Sch Fundamental Biol & Biotechnol, 79 Svobodny Pr, Krasnoyarsk 660041, Russia.
RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, Inst Biophys SB, 50-50 Akademgorodok, Krasnoyarsk 660036, Russia.
RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, LV Kirensky Inst Phys SB, 50-38 Akademgorodok, Krasnoyarsk 660036, Russia.
Russian Acad Sci, Siberian Branch, Krasnoyarsk Sci Ctr, Fed Res Ctr, 50 Akademgorodok, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Sch Petr & Gas Engn, 79 Svobodny Pr, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Volova, Tatiana G. G.; Prudnikova, Svetlana V. V.; Kiselev, Evgeniy G. G.; Nemtsev, Ivan V. V.; Vasiliev, Alexander D. D.; Kuzmin, Andrey P. P.; Shishatskaya, Ekaterina I. I.; Kiselev, Evgeniy; Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]

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