Myasoedova N.M.,Skryabin Institute of Biochemistry and Physiology of Microorganisms |
Gasanov N.B.,Pushchino State Institute of Natural science |
Chernykh A.M.,Skryabin Institute of Biochemistry and Physiology of Microorganisms |
Kolomytseva M.P.,Skryabin Institute of Biochemistry and Physiology of Microorganisms |
Golovleva L.A.,Skryabin Institute of Biochemistry and Physiology of Microorganisms
Applied Biochemistry and Microbiology | Year: 2015
The effects of a number of culture medium components, such as peptone, yeast extract, mono- and disaccharides, copper ions, 2,6-dimethylphenol, and polycaproamide fiber, on the laccase activity dynamics in the culture liquid and laccase isoform production by the Lentinus strigosus 1566 fungus were studied. It was demonstrated that some saccharides selectively induced or inhibited the synthesis of different laccase isoforms. Similar action was exerted by copper ions, 2,6-dimethylphenol, and polycaproamide fiber, as well as by their combination. Selective in vivo regulation of the production of certain laccase isoforms by basidial fungi by means of altering the culturing medium composition can be utilised for various biotechnological purposes. © Pleiades Publishing, Inc., 2015.
Methyloligella halotolerans gen. nov., sp. nov. and Methyloligella solikamskensis sp. nov., two non-pigmented halotolerant obligately methylotrophic bacteria isolated from the Ural saline environments
Doronina N.V.,Russian Academy of Sciences |
Poroshina M.N.,Pushchino State Institute of Natural science |
Kaparullina E.N.,Russian Academy of Sciences |
Ezhov V.A.,Russian Academy of Sciences |
Trotsenko Y.A.,Russian Academy of Sciences
Systematic and Applied Microbiology | Year: 2013
Two newly isolated halotolerant obligately methylotrophic bacteria (strains C2T and SK12T) with the serine pathway of C1 assimilation are described. The isolates are strictly aerobic, Gram negative, asporogenous, non-motile rods, forming rosettes, multiplying by binary fission. Mesophilic and neutrophilic, accumulate intracellularly compatible solute ectoine and poly-β-hydroxybutyrate. The novel strains are able to grow at 0 up to 16% NaCl (w/v), optimally at 3-5% NaCl. The major cellular fatty acids are C18:1ω7c and C19:0cyc and the prevailing quinone is Q-10. The predominant phospholipids are phosphatidylcholine, phosphatidylglycerol, phosphatidyldimethylethanolamine and phosphatidylethanolamine. Assimilate NH4 + by glutamate dehydrogenase and via the glutamate cycle (glutamine synthetase and glutamate synthase). The DNA G+C contents of strains C2T and SK12T are 60.9 and 60.5mol% (Tm), respectively. 16S rRNA gene sequence similarity between the two new isolates are 99% but below 94% with other members of the Alphaproteobacteria thus indicating that they can be assigned to a novel genus Methyloligella. Rather low level of DNA-DNA relatedness (53%) between the strains C2T and SK12T indicates that they represent two separate species of the new genus, for which the names Methyloligella halotolerans gen. nov., sp. nov. and Methyloligella solikamskensis sp. nov. are proposed. The type strain of Methyloligella halotolerans is C2T (=VKM B-2706T=CCUG 61687T=DSM 25045T) and the type strain of Methyloligella solikamskensis is SK12T (=VKM B-2707T=CCUG 61697T=DSM 25212T). © 2012 Elsevier GmbH.
Fedorov D.N.,Russian Academy of Sciences |
Zamakhaeva S.A.,Pushchino State Institute of Natural science |
Ezhov V.A.,Russian Academy of Sciences |
Doronina N.V.,Pushchino State Institute of Natural science |
Trotsenko Y.A.,Pushchino State Institute of Natural science
Applied Biochemistry and Microbiology | Year: 2014
The effect of the increased copy number of polyhydroxybutyrate (PHB) biosynthesis genes in pink-pigmented methylobacterium Methylobacterium extorquens G10 on properties of the biopolymer was studied. The activity of poly-3-hydroxybutyrate-synthase (PHB-synthase) was shown to increase and the molecular weight of synthesized PHB decreases twofold (150 → 79 kDa) after insertion of extra copies of phaC and phaCAB genes into cells of the producer strain, whereas the physicochemical properties of the plastic changed insignificantly. White mutant M. extorquens G10-W with disrupted synthesis of the carotenoid pigment (defect by the crtI gene, which codes for phytoene desaturase) was established to have the same rate of growth and level of PHB accumulation as the initial strain G10. The G10-W strain is a promising producer of PHB, with decreased expenses for purification and PHB biosynthesis. © 2014 Pleiades Publishing, Inc.
Shubina V.S.,Russian Academy of Sciences |
Shatalina Yu.V.,Pushchino State Institute of Natural science
Journal of Applied Spectroscopy | Year: 2013
We have used absorption spectroscopy to study the acid-base and metal-binding properties of two structurally similar flavanones: taxifolin and naringenin. We have determined the acid dissociation constants for taxifolin (pKa1 = 7.10 ± 0.05, pKa2 = 8.60 ± 0.09, pKa3 = 8.59 ± 0.19, pKa4 = 11.82 ± 0.36) and naringenin (pKa1 = 7.05 ± 0.05, pKa2 = 8.85 ± 0.09, pKa3 = 12.01 ± 0.38). The appearance of new absorption bands in the visible wavelength region let us determine the stoichiometric composition of the iron (II) complexes of the flavanones. We show that at pH 5, in solution there is a mixture of complexes between taxifolin and iron (II) ions in stoichiometric ratio 2:1 and 1:2, while at pH 7.4 and pH 9, we detect a 1:1 taxifolin:Fe(II) complex. We established that at these pH values, naringenin forms a 2:1 complex with iron (II) ions. We propose structures for the complexes formed. Comprehensive study of the acid-base properties and the metal-binding capability of the two structurally similar flavanones let us determine the structure-properties relation and the conditions under which antioxidant activity of the polyphenols appears, via chelation of variable-valence metal ions. © 2013 Springer Science+Business Media New York.
Kirkby N.S.,Queens Medical Research Institute |
Kirkby N.S.,Imperial College London |
Duthie K.M.,Queens Medical Research Institute |
Miller E.,Queens Medical Research Institute |
And 6 more authors.
Cardiovascular Research | Year: 2012
Aims The potent vasoconstrictor endothelin-1 (ET-1), acting on the endothelin-A (ETA) receptor, promotes intimal lesion formation following vascular injury. The endothelin-B (ETB) receptor, which mediates nitric oxide release and ET-1 clearance in endothelial cells, may moderate lesion formation, but this is less clear. We used selective ET receptor antagonists and cell-specific deletion to address the hypothesis that ETB receptors in the endothelium inhibit lesion formation following arterial injury. Methods and results Neointimal proliferation was induced by wire or ligation injury to the femoral artery in mice treated with selective ETA (ABT-627) and/or ETB antagonists (A192621). Measurement of lesion formation by optical projection tomography and histology indicated that ETA blockade reduced lesion burden in both models. Although ETB blockade had little effect on ligation injury-induced lesion formation, after wire injury, blockade of the ETB receptor increased lesion burden (184 of vehicle; P < 0.05) and reversed the protective effects of an ETA antagonist. Selective deletion of ETB receptors from the endothelium, however, had no effect on neointimal lesion size. Conclusion These results are consistent with ETB receptor activation playing an important role in limiting neointimal lesion formation following acute vascular injury, but indicate that this protective effect is not mediated by those ETB receptors expressed by endothelial cells. These data support the proposal that selective ETA antagonists may be preferable to mixed ETA/ETB antagonists for targeting the arterial response to injury. Published on behalf of the European Society of Cardiology. All rights reserved. © 2012 The Author.
Semenov M.V.,Moscow State University |
Stolnikova E.V.,Russian Academy of Sciences |
Ananyeva N.D.,Russian Academy of Sciences |
Ivashchenko K.V.,Pushchino State Institute of Natural science
Biology Bulletin | Year: 2013
The structure of the microbial community (the fungi-to-bacteria ratio) has been assessed by selective inhibition of the substrate-induced respiration (SIR) using streptomycin sulfate and cycloheximide antibiotics in the gray forest soil of eluvial, transite, transite-accumulative, and accumulative (meadow alluvial) parts of slope landscape on the right bank of the Oka River (near Pushchino, Moscow oblast) which represents an fallow, small-leaved wood, spruce forest, and meadow. The concentrations of bactericide and fungicide were selected experimentally for each landscape parts which provide the greatest SIR inhibition of the soil upon their individual application and in combination. Fungi were established to be predominant in the contribution to the total SIR which was found to be 82-97%. A dependence between the structure of the microbial community and the C/N ratio and pH of the soil was shown. © 2013 Pleiades Publishing, Ltd.
Agafonova N.V.,Pushchino State Institute of Natural science |
Doronina N.V.,Russian Academy of Sciences |
Trotsenko Y.A.,Russian Academy of Sciences
Applied Biochemistry and Microbiology | Year: 2016
The influence of colonization of the pea (Pisum sativum L.) by aerobic methylobacteria of five different species (Methylophilus flavus Ship, Methylobacterium extorquens G10, Methylobacillus arboreus Iva, Methylopila musalis MUSA, Methylopila turkiensis Side1) on plant resistance to paraquat-induced stresses has been studied. The normal conditions of pea colonization by methylobacteria were characterized by a decrease in the activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidases) and in the concentrations of endogenous H2O2, proline, and malonic dialdehyde, which is a product of lipid peroxidation and indicator of damage to plant cell membranes, and an increase in the activity of the photosynthetic apparatus (the content of chlorophylls а, b and carotenoids). In the presence of paraquat, the colonized plants had higher activities of antioxidant enzymes, stable photosynthetic indices, and a less intensive accumulation of the products of lipid peroxidation as compared to noncolonized plants. Thus, colonization by methylobacteria considerably increased the adaptive protection of pea plants to the paraquat-induced oxidative stress. © 2016, Pleiades Publishing, Inc.
PubMed | Pushchino State Institute of Natural science, Russian Academy of Sciences, Pushchino State University and RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry
Type: Journal Article | Journal: FEBS letters | Year: 2015
The bacterial cell wall muramyl dipeptides MDP and glucosaminyl-MDP (GMDP) are powerful immunostimulators but their binding target remains controversial. We previously reported expression cloning of GMDP-binding polypeptides and identification of Y-box protein 1 (YB-1) as their sole target. Here we show specific binding of GMDP to recombinant YB-1 protein and subcellular colocalization of YB-1 and GMDP. GMDP binding to YB-1 upregulated gene expression levels of NF-B2, a mediator of innate immunity. Furthermore, YB-1 knockdown abolished GMDP-induced Nfkb2 expression. GMDP/YB-1 stimulation led to NF-B2 cleavage, transport of activated NF-B2 p52 to the nucleus, and upregulation of NF-B2-dependent chemokine Cxcr4 gene expression. Therefore, our findings identify YB-1 as new target for muramyl peptide signaling.
PubMed | Pushchino State Institute of Natural science and Russian Academy of Sciences
Type: Journal Article | Journal: Microorganisms | Year: 2016
We have expressed the l-malate dehydrogenase (MDH) genes from aerobic methanotrophs Methylomicrobium alcaliphilum 20Z and Methylosinus trichosporium OB3b as his-tagged proteins in Escherichia coli. The substrate specificities, enzymatic kinetics and oligomeric states of the MDHs have been characterized. Both MDHs were NAD-specific and thermostable enzymes not affected by metal ions or various organic metabolites. The MDH from M. alcaliphilum 20Z was a homodimeric (2 35 kDa) enzyme displaying nearly equal reductive (malate formation) and oxidative (oxaloacetate formation) activities and higher affinity to malate (Km = 0.11 mM) than to oxaloacetate (Km = 0.34 mM). The MDH from M. trichosporium OB3b was homotetrameric (4 35 kDa), two-fold more active in the reaction of oxaloacetate reduction compared to malate oxidation and exhibiting higher affinity to oxaloacetate (Km = 0.059 mM) than to malate (Km = 1.28 mM). The kcat/Km ratios indicated that the enzyme from M. alcaliphilum 20Z had a remarkably high catalytic efficiency for malate oxidation, while the MDH of M. trichosporium OB3b was preferable for oxaloacetate reduction. The metabolic roles of the enzymes in the specific metabolism of the two methanotrophs are discussed.
PubMed | Pushchino State Institute of Natural science and RAS Engelhardt Institute of Molecular Biology
Type: Journal Article | Journal: Molekuliarnaia biologiia | Year: 2015
The aim of this study was to estimate the efficacy of mesenchymal stem cell-based suicide gene therapy in mice bearing murine melanoma B16F10. Adipose mesenchymal stem cells (MSCs) were transfected with plasmid constructs expressing cytosine deaminase fused with uracil phosphoribosyltransferase (CDA/UPRT) or CDA/UPRT fused with HSV-1 tegument protein VP22 (CDA/UPRT/VP22). In this study, we demonstrate that direct intratumoral transplantation of MSCs expressing CDA/UPRT or CDA/UPRT/VP22 followed by systemic administration of 5-fluorocytosine (5-FC) results in a significant inhibition of tumor growth. There was a 53% reduction in tumor volume in mice treated with CDA/UPRT-MSCs and 58% reduction in tumor volume in mice treated with CDA/UPRT/VP22-MSCs as compared with control animals transplanted with B16F10 melanoma alone. Injection of CDA/UPRT-MSC and CDA/UPRT/VP22-MSC prolonged the life span of mice bearing B16F10 melanoma by 15 and 26%, respectively. The data indicate that in murine B16F10 melanoma model, MSCs encoding CDA/UPRT suicide gene have a significant antitumor effect.