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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : 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.
Заглавие : Bacterial Cellulose (BC) and BC Composites: Production and Properties
Колич.характеристики :24 с
Коллективы : Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]
Место публикации : Nanomaterials: MDPI, 2022. - Vol. 12, Is. 2. - Ст.192. - ISSN 2079-4991(eISSN), DOI 10.3390/nano12020192
Примечания : 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.
Предметные рубрики: SILVER NANOPARTICLES
GLUCONACETOBACTER-HANSENII
MICROBIAL CELLULOSE
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.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Volova, Tatiana G., Kiselev, Evgeniy G., Demidenko, Alexey V., Zhila, Natalia O., Nemtsev, Ivan V., Lukyanenko, Anna V.
Заглавие : Production and Properties of Microbial Polyhydroxyalkanoates Synthesized from Hydrolysates of Jerusalem Artichoke Tubers and Vegetative Biomass
Колич.характеристики :25 с
Коллективы : Project "Agropreparations of the new generation: a strategy of construction and realization" [075-15-2021-626]; State Assignment of the Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]
Место публикации : Polymers: MDPI, 2022. - Vol. 14, Is. 1. - Ст.132. - ISSN 2073-4360(eISSN), DOI 10.3390/polym14010132
Примечания : Cited References:93. - This study 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" (polymer synthesis, properties), and by the State Assignment of the Ministry of Science and Higher Education of the Russian Federation No. FSRZ-2020-0006 (films production, surface properties).
Предметные рубрики: GLUCOSE-UTILIZING STRAIN
RALSTONIA-EUTROPHA
ASPERGILLUS-NIGER
ACID
Аннотация: One of the major challenges in PHA biotechnology is optimization of biotechnological processes of the entire synthesis, mainly by using new inexpensive carbon substrates. A promising substrate for PHA synthesis may be the sugars extracted from the Jerusalem artichoke. In the present study, hydrolysates of Jerusalem artichoke (JA) tubers and vegetative biomass were produced and used as carbon substrate for PHA synthesis. The hydrolysis procedure (the combination of aqueous extraction and acid hydrolysis, process temperature and duration) influenced the content of reducing substances (RS), monosaccharide contents, and the fructose/glucose ratio. All types of hydrolysates tested as substrates for cultivation of three strains-C. necator B-10646 and R. eutropha B 5786 and B 8562-were suitable for PHA synthesis, producing different biomass concentrations and polymer contents. The most productive process, conducted in 12-L fermenters, was achieved on hydrolysates of JA tubers (X = 66.9 g/L, 82% PHA) and vegetative biomass (55.1 g/L and 62% PHA) produced by aqueous extraction of sugars at 80 degrees C followed by acid hydrolysis at 60 degrees C, using the most productive strain, C. necator B-10646. The effects of JA hydrolysates on physicochemical properties of PHAs were studied for the first time. P(3HB) specimens synthesized from the JA hydrolysates, regardless of the source (tubers or vegetative biomass), hydrolysis conditions, and PHA producing strain employed, exhibited the 100-120 degrees C difference between the T-melt and T-degr, prevailing of the crystalline phase over the amorphous one (C-x between 69 and 75%), and variations in weight average molecular weight (409-480) kDa. Supplementation of the culture medium of C. necator B-10646 grown on JA hydrolysates with potassium valerate and epsilon-caprolactone resulted in the synthesis of P(3HB-co-3HV) and P(3HB-co-4HB) copolymers that had decreased degrees of crystallinity and molecular weights, which influenced the porosity and surface roughness of polymer films prepared from them. The study shows that JA hydrolysates used as carbon source enabled productive synthesis of PHAs, comparable to synthesis from pure sugars. The next step is to scale up PHA synthesis from JA hydrolysates and conduct the feasibility study. The present study contributes to the solution of the critical problem of PHA biotechnology-finding widely available and inexpensive substrates.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Tomilin F. N., Rogova A. V., Burakova L. P., Tchaikovskaya O. N., Avramov P. V., Fedorov D. G., Vysotski E. S.
Заглавие : Unusual shift in the visible absorption spectrum of an active ctenophore photoprotein elucidated by time-dependent density functional theory
Место публикации : Photochem. Photobiol. Sci.: Springer Nature, 2021. - Vol. 20, Is. 4. - С. 559-570. - ISSN 1474905X (ISSN), DOI 10.1007/s43630-021-00039-5
Аннотация: Active hydromedusan and ctenophore Ca2+-regulated photoproteins form complexes consisting of apoprotein and strongly non-covalently bound 2-hydroperoxycoelenterazine (an oxygenated intermediate of coelenterazine). Whereas the absorption maximum of hydromedusan photoproteins is at 460–470 nm, ctenophore photoproteins absorb at 437 nm. Finding out a physical reason for this blue shift is the main objective of this work, and, to achieve it, the whole structure of the protein–substrate complex was optimized using a linear scaling quantum–mechanical method. Electronic excitations pertinent to the spectra of the 2-hydroperoxy adduct of coelenterazine were simulated with time-dependent density functional theory. The dihedral angle of 60° of the 6-(p-hydroxy)-phenyl group relative to the imidazopyrazinone core of 2-hydroperoxycoelenterazine molecule was found to be the key factor determining the absorption of ctenophore photoproteins at 437 nm. The residues relevant to binding of the substrate and its adopting the particular rotation were also identified. © 2021, The Author(s), under exclusive licence to European Photochemistry Association,European Society for Photobiology.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Ismagilov, Rinat, Malykhin, Sergei, Puzyr, Aleksey, Loginov, Artem, Kleshch, Victor, Obraztsov, Alexander
Заглавие : Single-Crystal Diamond Needle Fabrication Using Hot-Filament Chemical Vapor Deposition
Колич.характеристики :9 с
Коллективы : Russian Science FoundationRussian Science Foundation (RSF) [19-79-00203]; Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [18-29-19071]
Место публикации : Materials: MDPI, 2021. - Vol. 14, Is. 9. - Ст.2320. - ISSN 1996-1944(eISSN), DOI 10.3390/ma14092320
Примечания : Cited References:32. - This work was supported by the Russian Science Foundation (project no. 19-79-00203) and by the Russian Foundation for Basic Research (grant no. 18-29-19071, in part for PL and Raman inspection).
Аннотация: Single-crystal diamonds in the form of micrometer-scale pyramids were produced using a combination of hot-filament (HF) chemical vapor deposition (CVD) and thermal oxidation processes. The diamond pyramids were compared here with similar ones that were manufactured using plasma-enhanced (PE) CVD. The similarities revealed in the morphology, Raman, and photoluminescent characteristics of the needles obtained using the hot-filament and plasma-enhanced CVD are discussed in connection with the diamond film growth mechanism. This work demonstrated that the HF CVD method has convincing potential for the fabrication of single-crystal diamond needles in the form of regularly shaped pyramids on a large surface area, even on non-conducting substrates. The experimental results demonstrated the ability for the mass production of the single-crystal needle-like diamonds, which is important for their practical application.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kiselev E. G., Vasiliev A. D., Volova T. G.
Заглавие : Synthesis and characterization of multicomponent PHAs
Место публикации : J. Sib. Fed. Univ. - Biol.: Siberian Federal University, 2021. - Vol. 14, Is. 1. - С. 97-113. - ISSN 19971389 (ISSN), DOI 10.17516/1997-1389-0325
Примечания : Cited By :1
Аннотация: Cupriavidus necator B10646 bacterial cells were cultivated in the mode of synthesis of the reserve polyhydroxyalkanoates (PHAs) in the growth medium that contained, in addition to glucose as the main substrate, precursor substrates of the monomers of various monocarboxylic acids - salts of valeric and hexanoic acids, propionate, and ?-butyrolactone. PHA terpolymers and quaterpolymers with different compositions and proportions of monomers were synthesized, and their physicochemical properties were studied. The terpolymers were composed of monomers of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 4-hydroxybutyrate (4HB), or 3-hydroxyhexanoate (3HHx) and had the following compositions: P(3HB/3HB/4HHx) and P(3HB/3HV/4HHx). The quaterpolymers had the following composition: P(3HB/3HV/4HB/3HHx). All copolymer samples, regardless of the composition and proportions of monomers, had lower molecular weights and higher polydispersity values compared to the highly crystalline 3-hydroxybutyrate homopolymer, but retained thermal stability properties, with a difference between the melting point and thermal degradation of at least 100-110 °C. The inclusion of 3HV, 4HB, and 3HHx monomers in the C-chain of 3HB caused changes in the crystalline to amorphous phase ratio and a significant decrease in the degree of crystallinity (Cx), which depended on the type of monomers and their contents in the copolymer. The maximum decrease in Cx (9-17 %) was detected in the P(3HB/3HV/4HB) terpolymer and the P(3HB/3HV/4HB/3HHx) quaterpolymer (30-36 %). The study confirms that there is the possibility of synthesizing polymers with various compositions, including new ones, which differ significantly in their basic properties. © Siberian Federal University. All rights reserved.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E., Nemtsev I., Lukyanenko A., Vasiliev A., Kiselev E., Sukovatyi A., Volova T.
Заглавие : Polymer Films of Poly-3-hydroxybutyrate Synthesized by Cupriavidus necator from Different Carbon Sources
Место публикации : J. Polym. Environ.: Springer, 2021. - Vol. 29, Is. 3. - С. 837-850. - ISSN 15662543 (ISSN), DOI 10.1007/s10924-020-01924-3
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.
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Вид документа : Статья из сборника (выпуск продолж. издания)
Шифр издания :
Автор(ы) : Kiselev E. G., Zhila N. O., Volova T. G.
Заглавие : Biodegradable polymers - Perspectives and applications in agriculture
Место публикации : 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
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.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Zhila, Natalia O., Sapozhnikova, Kristina Yu, Kiselev, Evgeniy G., Vasiliev, Alexander D., Nemtsev, Ivan, V, Shishatskaya, Ekaterina, I, Volova, Tatiana G.
Заглавие : Properties of Degradable Polyhydroxyalkanoates (PHAs) Synthesized by a New Strain, Cupriavidus necator IBP/SFU-1, from Various Carbon Sources
Колич.характеристики :19 с
Коллективы : 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]
Место публикации : Polymers: MDPI, 2021. - Vol. 13, Is. 18. - Ст.3142. - ISSN 2073-4360(eISSN), DOI 10.3390/polym13183142
Примечания : 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).
Предметные рубрики: RALSTONIA-EUTROPHA
POLY(3-HYDROXYBUTYRATE) PRODUCTION
PLANT OIL
ACID
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.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Chen S. -F., Vysotski E. S., Liu Y. -J.
Заглавие : H2O-Bridged Proton-Transfer Channel in Emitter Species Formation in Obelin Bioluminescence
Место публикации : J Phys Chem B: American Chemical Society, 2021. - Article in press. - ISSN 15206106 (ISSN), DOI 10.1021/acs.jpcb.1c03985
Аннотация: Bioluminescence of a number of marine organisms is conditioned by Ca2+-regulated photoprotein (CaRP) with coelenterazine as the reaction substrate. The reaction product, coelenteramide, at the first singlet excited state (S1) is the emitter of CaRP. The S1-state coelenteramide is produced via the decomposition of coelenterazine dioxetanone. Experiments suggested that the neutral S1-coelenteramide is the primary emitter species. This supposition contradicts with theoretical calculations showing that the anionic S1-coelenteramide is a primary product of the decomposition of coelenterazine dioxetanone. In this study, applying molecular dynamic (MD) simulations and the hybrid quantum mechanics/molecular mechanics (QM/MM) method, we investigated a proton-transfer (PT) process taking place in CaRP obelin from Obelia longissima for emitter formation. Our calculations demonstrate a concerted PT process with a water molecule as a bridge between anionic S1-coelenteramide and the nearest histidine residue. The low activation barrier as well as the strong hydrogen-bond network between the proton donor and the proton acceptor suggests a fast PT process comparable with that of the lifetime of excited anionic S1-coelenteramide. The existence of the PT process eliminates the discrepancy between experimental and theoretical studies. The fast PT process at emitter formation can also take place in other CaRPs. © 2021 American Chemical Society.
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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Zhila N. O., Sapozhnikova K. Yu., Kiselev E. G., Vasiliev A. D., Nemtsev I. V., Shishatskaya E. I., Volova T. G.
Заглавие : Properties of degradable polyhydroxyalkanoates (Phas) synthesized by a new strain, cupriavidus necator ibp/sfu-1, from various carbon sources
Место публикации : Polym.: MDPI, 2021. - Vol. 13, Is. 18. - Ст.3142. - ISSN 20734360 (ISSN), DOI 10.3390/polym13183142
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.
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11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lisitsa A. E., Sukovatyi L. A., Bartsev S. I., Deeva A. A., Kratasyuk V. A., Nemtseva E. V.
Заглавие : Mechanisms of viscous media effects on elementary steps of bacterial bioluminescent reaction
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2021. - Vol. 22, Is. 16. - Ст.8827. - ISSN 16616596 (ISSN), DOI 10.3390/ijms22168827
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.
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12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lisitsa, Albert E., Sukovatyi, Lev A., Bartsev, Sergey, I, Deeva, Anna A., Kratasyuk, Valentina A., Nemtseva, Elena, V
Заглавие : Mechanisms of Viscous Media Effects on Elementary Steps of Bacterial Bioluminescent Reaction
Колич.характеристики :19 с
Коллективы : 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]
Место публикации : Int. J. Mol. Sci.: MDPI, 2021. - Vol. 22, Is. 16. - Ст.8827. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms22168827
Примечания : 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.
Предметные рубрики: FLAVIN INTERMEDIATE
REDUCED FLAVIN
RATE CONSTANTS
LUCIFERASE
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.
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13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva E. V., Jiang T., Malikova N. P., Li M., Vysotski E. S.
Заглавие : Bioluminescent properties of semi-synthetic obelin and aequorin activated by coelenterazine analogues with modifications of C-2, C-6, and C-8 substituents
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2020. - Vol. 21, Is. 15. - Ст.5446. - С. 1-21. - ISSN 16616596 (ISSN), DOI 10.3390/ijms21155446
Аннотация: Ca2+-regulated photoproteins responsible for bioluminescence of a variety of marine organisms are single-chain globular proteins within the inner cavity of which the oxygenated coelenterazine, 2-hydroperoxycoelenterazine, is tightly bound. Alongside with native coelenterazine, photoproteins can also use its synthetic analogues as substrates to produce flash-type bioluminescence. However, information on the effect of modifications of various groups of coelenterazine and amino acid environment of the protein active site on the bioluminescent properties of the corresponding semi-synthetic photoproteins is fragmentary and often controversial. In this paper, we investigated the specific bioluminescence activity, light emission spectra, stopped-flow kinetics and sensitivity to calcium of the semi-synthetic aequorins and obelins activated by novel coelenterazine analogues and the recently reported coelenterazine derivatives. Several semi-synthetic photoproteins activated by the studied coelenterazine analogues displayed sufficient bioluminescence activities accompanied by various changes in the spectral and kinetic properties as well as in calcium sensitivity. The poor activity of certain semi-synthetic photoproteins might be attributed to instability of some coelenterazine analogues in solution and low efficiency of 2-hydroperoxy adduct formation. In most cases, semi-synthetic obelins and aequorins displayed different properties upon being activated by the same coelenterazine analogue. The results indicated that the OH-group at the C-6 phenyl ring of coelenterazine is important for the photoprotein bioluminescence and that the hydrogen-bond network around the substituent in position 6 of the imidazopyrazinone core could be the reason of different bioluminescence activities of aequorin and obelin with certain coelenterazine analogues. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Volova T., Sapozhnikova K., Zhila N.
Заглавие : Cupriavidus necator B-10646 growth and polyhydroxyalkanoates production on different plant oils
Место публикации : Int. J. Biol. Macromol.: Elsevier B.V., 2020. - Vol. 164. - С. 121-130. - ISSN 01418130 (ISSN), DOI 10.1016/j.ijbiomac.2020.07.095
Аннотация: The study addresses the growth of the wild-type strain Cupriavidus necator B-10646 and synthesis of polyhydroxyalkanoates by this strain on media containing plant oils with different compositions of fatty acids: palm, Siberian oilseed, and refined and unrefined sunflower seed oils. The study showed that the best carbon substrate was palm oil. Comparison of fatty acid compositions of the starting oils and unutilized residual substrates showed that C. necator B-10646 cells consumed the fatty acids from palm oil evenly while in experiments with other oils, they utilized polyenoic fatty acids first. Higher production parameters of the culture were obtained by preparation of emulsified oil medium using Tween 80 and sodium cocoyl glutamate as emulsifiers. All polyhydroxyalkanoate specimens were terpolymers that contained 3-hydroxybutyrate as the major component and minor amounts of 3-hydroxyvalerate (0.9–1.9 mol%) and 3-hydroxyhexanoate (0.5–1.1 mol%). Molecular weight of polyhydroxyalkanoate specimens depended on the type of plant oil and emulsifier. © 2020
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15.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva, Elena, V, Jiang, Tianyu, Malikova, Natalia P., Li, Minyong, Vysotski, Eugene S.
Заглавие : Bioluminescent Properties of Semi-Synthetic Obelin and Aequorin Activated by Coelenterazine Analogues with Modifications of C-2, C-6, and C-8 Substituents
Колич.характеристики :21 с
Коллективы : RFBRRussian Foundation for Basic Research (RFBR); NSFCNational Natural Science Foundation of China (NSFC) [20-54-53011]; Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [18-44-242001]; Krasnoyarsk Regional Fund of Science; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81874308]; Shandong Natural Science FoundationNatural Science Foundation of Shandong Province [ZR2018ZC0233]; Government of Krasnoyarsk Territory
Место публикации : Int. J. Mol. Sci.: MDPI, 2020. - Vol. 21, Is. 15. - Ст.5446. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms21155446
Примечания : Cited References:50. - The reported study was funded by RFBR and NSFC according to the research project No. 20-54-53011 (E.V.E. and N.P.M.), Russian Foundation for Basic Research (No. 18-44-242001), Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science (E.S.V.), the National Natural Science Foundation of China (No. 81874308), and the Shandong Natural Science Foundation (No. ZR2018ZC0233) (M.L.).
Предметные рубрики: CA2+-REGULATED PHOTOPROTEINS
SPECTROSCOPIC PROPERTIES
Аннотация: Ca2+-regulated photoproteins responsible for bioluminescence of a variety of marine organisms are single-chain globular proteins within the inner cavity of which the oxygenated coelenterazine, 2-hydroperoxycoelenterazine, is tightly bound. Alongside with native coelenterazine, photoproteins can also use its synthetic analogues as substrates to produce flash-type bioluminescence. However, information on the effect of modifications of various groups of coelenterazine and amino acid environment of the protein active site on the bioluminescent properties of the corresponding semi-synthetic photoproteins is fragmentary and often controversial. In this paper, we investigated the specific bioluminescence activity, light emission spectra, stopped-flow kinetics and sensitivity to calcium of the semi-synthetic aequorins and obelins activated by novel coelenterazine analogues and the recently reported coelenterazine derivatives. Several semi-synthetic photoproteins activated by the studied coelenterazine analogues displayed sufficient bioluminescence activities accompanied by various changes in the spectral and kinetic properties as well as in calcium sensitivity. The poor activity of certain semi-synthetic photoproteins might be attributed to instability of some coelenterazine analogues in solution and low efficiency of 2-hydroperoxy adduct formation. In most cases, semi-synthetic obelins and aequorins displayed different properties upon being activated by the same coelenterazine analogue. The results indicated that the OH-group at the C-6 phenyl ring of coelenterazine is important for the photoprotein bioluminescence and that the hydrogen-bond network around the substituent in position 6 of the imidazopyrazinone core could be the reason of different bioluminescence activities of aequorin and obelin with certain coelenterazine analogues.
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16.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Volova, Tatiana, Sapozhnikova, Kristina, Zhila, Natalia
Заглавие : Cupriavidus necator B-10646 growth and polyhydroxyalkanoates production on different plant oils
Колич.характеристики :10 с
Коллективы : Project "Agropreparations of the new generation: a strategy of construction and realization" [074-02-2018-328]; Government of the Russian Federation; State assignment of the Ministry of Science and Higher Education of the Russian Federation [FSRZ-2020-0006]
Место публикации : Int. J. Biol. Macromol.: ELSEVIER, 2020. - Vol. 164. - С. 121-130. - ISSN 0141-8130, DOI 10.1016/j.ijbiomac.2020.07.095. - ISSN 1879-0003(eISSN)
Примечания : Cited References:52. - This studywas financially supported by Project "Agropreparations of the new generation: a strategy of construction and realization" (Agreement No 074-02-2018-328) in accordance with Resolution No 220 of the Government of the Russian Federation of April 9, 2010, "On measures designed to attract leading scientists to the Russian institutions of higher learning" (polymer synthesis fromplant oils), and by the State assignment of the Ministry of Science and Higher Education of the Russian Federation No. FSRZ-2020-0006 (polymer properties).
Предметные рубрики: FATTY-ACID-COMPOSITION
PHA SYNTHASE GENE
PALM KERNEL OIL
Аннотация: The study addresses the growth of the wild-type strain Cupriavidus necator B-10646 and synthesis of polyhydroxyalkanoates by this strain on media containing plant oils with different compositions of fatty acids: palm, Siberian oilseed, and refined and unrefined sunflower seed oils. The study showed that the best carbon substrate was palm oil. Comparison of fatty acid compositions of the starting oils and unutilized residual substrates showed that C. necator B-10646 cells consumed the fatty acids from palm oil evenly while in experiments with other oils, they utilized polyenoic fatty acids first. Higher production parameters of the culture were obtained by preparation of emulsified oil medium using Tween 80 and sodium cocoyl glutamate as emulsifiers. All polyhydroxyalkanoate specimens were terpolymers that contained 3-hydroxybutyrate as the major component and minor amounts of 3-hydroxyvalerate (0.9-1.9 mol%) and 3-hydroxyhexanoate (0.5-1.1 mol%). Molecular weight of polyhydroxyalkanoate specimens depended on the type of plant oil and emulsifier. (C) 2020 Elsevier B.V. All rights reserved.
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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya, Ekaterina, Nemtsev, Ivan, Lukyanenko, Anna, Vasiliev, Alexander, Kiselev, Evgeniy, Sukovatyi, Aleksey, Volova, Tatiana
Заглавие : Polymer Films of Poly-3-hydroxybutyrate Synthesized byCupriavidus necatorfrom Different Carbon Sources
Колич.характеристики :14 с
Коллективы : 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]
Место публикации : J. Polym. Environ.: SPRINGER, 2020. - Article in press. - ISSN 1566-2543, DOI 10.1007/s10924-020-01924-3. - ISSN 1572-8919(eISSN)
Примечания : 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.
Предметные рубрики: CHEMOMECHANICAL PROPERTIES
RALSTONIA-EUTROPHA
SURFACE-ROUGHNESS
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.
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva, Elena V., Vysotski, Eugene S.
Заглавие : Exploring Bioluminescence Function of the Ca2+-regulated Photoproteins with Site-directed Mutagenesis
Колич.характеристики :16 с
Коллективы : Russian Foundation for Basic Research [17-04-00764]; Russian Academy of Sciences [0356-2017-0017]
Место публикации : Photochem. Photobiol.: WILEY, 2019. - Vol. 95, Is. 1. - С. 8-23. - ISSN 0031-8655, DOI 10.1111/php.12945. - ISSN 1751-1097(eISSN)
Примечания : Cited References:88. - This work was supported by grant 17-04-00764 of Russian Foundation for Basic Research and the state budgetallocated to the fundamental research at the Russian Academy of Sciences (project 0356-2017-0017).
Предметные рубрики: CALCIUM-BINDING PHOTOPROTEIN
GREEN-FLUORESCENT PROTEIN
JELLYFISH
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--aequorin--obelin--clytin--mitrocomin--ef-hand protein
Аннотация: Site-directed mutagenesis is a powerful tool to investigate the structure-function relationship of proteins and a function of certain amino acid residues in catalytic conversion of substrates during enzymatic reactions. Hence, it is not surprising that this approach was repeatedly applied to elucidate the role of certain amino acid residues in various aspects of photoprotein bioluminescence, mostly for aequorin and obelin, and to design mutant photoproteins with altered properties (modified calcium affinity, faster or slower bioluminescence kinetics, different emission color) which would either allow the development of novel bioluminescent assays or improvement of characteristics of the already existing ones. This information, however, is scattered over different articles. In this review, we systematize the findings that were made using site-directed mutagenesis studies regarding the impact of various amino acid residues on bioluminescence of hydromedusan Ca2+-regulated photoproteins. All key residues that have been identified are pinpointed, and their influence on different aspects of photoprotein functioning such as active photoprotein complex formation, bioluminescence reaction, calcium response and light emitter formation is discussed.
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19.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Menzyanova N. G., Pyatina S. А., Nikolaeva E. D., Shabanov A. V., Nemtsev I. V., Stolyarov D. P., Dryganov D. B., Sakhnov E. V., Shishatskaya E. I.
Заглавие : Screening of biopolymeric materials for cardiovascular surgery toxicity—Evaluation of their surface relief with assessment of morphological aspects of monocyte/macrophage polarization in atherosclerosis patients
Место публикации : Toxicol. Rep.: Elsevier Inc., 2019. - Vol. 6. - С. 74-90. - ISSN 22147500 (ISSN) , DOI 10.1016/j.toxrep.2018.11.009
Ключевые слова (''Своб.индексиров.''): atherosclerosis--cell morphology--intravascular stenting--macrophages--monocytes--polyhydroxyalkanoates
Аннотация: The morphotypes of human macrophages (MPh) were studied in the culture on nano-structured biopolymer substrates, made from polyhydroxyalcanoates (PHAs) of five various monomer compositions, followed by the solvent evaporation. Its surface relief, which was further in direct contact with human cells in vitro, was analyzed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). It was shown, that the features of the micro/nano relief depend on the monomeric composition of the polymer substrates. Monocytes (MN) of patients with atherosclerosis and cardiac ischemia, undergoing stenting and conventional anti-atherosclerotic therapy, were harvested prior and after stenting. MN were isolated and cultured, with the transformation into MPh in direct contact with biopolymer culture substrates with different monomer composition and nano-reliefs, and transformed into MPh, in comparison with the same process on standard culture plastic. Sub-populations of cells with characteristic morphology in each phenotypic class were described, and their quantitative ratios for each sample of polymers were counted as an intermediate result in the development of “smart” material for cardiovascular devices. The results obtained allow us to assume, that the processes of MPh differentiation and polarization in vitro depend not only on the features of the micro/nano relief of biopolymer substrates, but also on the initial state of MN in vivo and general response of patients. © 2018 The Authors
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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Volova T., Kiselev E., Zhila N., Shishatskaya E.
Заглавие : Synthesis of Polyhydroxyalkanoates by Hydrogen-Oxidizing Bacteria in a Pilot Production Process
Место публикации : Biomacromolecules: American Chemical Society, 2019. - Article in press. - ISSN 15257797 (ISSN) , DOI 10.1021/acs.biomac.9b00295
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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