


Vol 54, No 1 (2018)
- Year: 2018
- Articles: 13
- URL: https://journal-vniispk.ru/0003-6838/issue/view/9113
Article
Regulation of Escherichia coli Biofilm Formation (Review)
Abstract
The review considers the regulatory mechanisms controlling the formation of biofilms by bacteria Escherichia coli. Under harsh conditions, microbial populations transfer to the structured mode of existence by building biofilms. The regulation of biofilm formation is a complex multistage process. Environmental signals are perceived by two-component signaling systems, which fulfill their transduction to the genome. This switches microbial cells from the planktonic motile lifestyle to the sessile.



Study of Structural-Functional Organization of Nucleoside Phosphorylases of Gammaproteobacteria. Special Aspects of Functioning of Uridine Phosphorylase Phosphate-Binding Site
Abstract
Series of mutant genes of prokaryotic uridine phosphorylases (Shewanella oneidensis MR-1, Escherichia coli) were constructed, and the resulting strains-producers of the corresponding proteins were obtained. Proteins were purified, and their physicochemical and fermentative properties were studied. On the basis of the obtained data, the role of individual amino acid residues of the polypeptide chain of uridine phosphorylases in the formation and functioning of the phosphate-binding site in these proteins was shown. The assumption of independent binding of two substrates, ion of inorganic phosphate (Pi) and uridine (Urd), by nucleoside phosphorylases, was made.



Vanillin Resistance Induced by BssS Overexpression in Escherichia coli
Abstract
Overexpression of the BssS gene, a biofilm formation regulator, in planktonic Escherichia coli cells has been shown to confer the vanillin-resistant phenotype Vanr to the bacteria. The MG1655PL-tac-bssS strain started growing in liquid aerated LB medium with 2 g/L vanillin after a lag phase of 17 ± 2 h, whereas the original MG1655 strain did not grow under these conditions. The role of aldehyde reductase YqhD, a vanillin- degrading enzyme, in Vanr phenotype formation has been assessed. However, the Vanr trait in the MG1655PL-tac-bssS strain primarily depended on autoinducer-2 (AI-2), which formed in E. coli cells with an intact luxS gene. We supposed that BssS acts together with autoinducer-2 (which presumably accumulated during the prolonged lag phase) to induce vanillin resistance determined by changes in the expression of a range of genes.



Methemoglobin Reductase of Bacteria and Bacteroids Bradyrhizobium lupini: Purification and Properties
Abstract
Reductase capable of reducing hemoglobin-like proteins was isolated from nodule bacteria Bradyrhizobium lupini and bacteroids of lupine root nodules. It is similar in some properties to many known methemoglobin reductases reducing animal and plant hemoglobins. It is a NADH-dependent FAD-containing flavoprotein with molecular weight of 87 kDa without metals. The presence of such enzymes in prokaryotes could be an explanation for the physiological activity of both bacterial and eukaryotic hemoglobins expressed in bacterial cells.



Isolation and Properties of Succinate Dehydrogenase Isozymes from Maize Scutellum (Zea mays L.)
Abstract
Four electrophoretically homogeneous succinate dehydrogenase isoenzymes with specific activities of 0.041, 0.110, 0.030 and 0.037 U/mg protein were isolated and purified from maize scutellum (Zea mays L.). Their catalytic properties and pH optimums were studied.



Catalytic Properties of Lipase Entrapped as Lysates of Recombinant Strain-Producer rEscherichia coli/lip into Nanocarbon-in-Silica Composites in the Bioconversion of Triglycerides and Fatty Acids
Abstract
Composite multi-component biocatalysts were prepared by entrapping lysates of a recombinant rE. coli/lip strain producing Thermomyces lanuginosus lipase into composite nanocarbon-containing matrices based on a SiO2 xerogel. The dependence of the lipase activity and operational stability on the type of the carbon component (nanotubes or nanospheres of different diameters) was studied in the bioconversion of triglycerides (hydrolysis and interesterification), as well as in the esterification of saturated fatty acids—namely, butyric (C4:0), capric (C10:0), and stearic (C18:0) acids—with isoamyl alcohol. It was shown that the biocatalytic properties were determined by both the texture parameters of the nanostructured carbon included and the type of enzymatic reaction performed. Biocatalysts without a nanocarbon component had the highest operational stability in the batch process of interesterification of sunflower oil with ethyl acetate; the half-life time was found to be 720 h at 40°C. Biocatalysts containing carbon nanotubes of ~21 nm in diameter were five to six times more active in the batch esterification process than biocatalysts without a nanocarbon component. Biocatalysts containing carbon nanotubes catalyzed the synthesis of esters in a binary organic solvent (hexane and diethyl ether) without a loss of activity for more than 500 h at 40°C.



Nitrocellulose Degradation by the Fungus Fusarium solani
Abstract
The degradation of native and pretreated nitrocellulose (NC) by the microscopic fungus Fusarium solani VKM F-819 and a mixed culture of the fungus with a sulfate-reducing bacterium Desulfovibrio desulfuricans VKM B-1388 has been studied. It has been shown that NC pretreatment with UV radiation and ozone promoted its subsequent biodegradation. The degradation of the thus treated NC by a mixed culture of F. solani and D. desulfuricans was the most effective as compared to all other treatment options. The NC nitrogen content decreased from 13.38 to 10.03%; the number average (Mn) and weight average (Mw) molecular masses decreased by three and two times, respectively. These magnitudes were achieved after 5 days of incubation of the pretreated NC. The obtained data can be used to further develop NC degradation technology.



The Influence of Chemical Mutagenesis on the Properties of the Cyclosporine a High-Producer Strain Tolypocladium inflatum VKM F-3630D
Abstract
The effects of treatment with the chemical mutagen N-nitroso-N-methylurea (NMU) on the survival rate, morphological variability, and the level of production of cyclosporine A (CyA) in the Tolypocladium inflatum subsporum blastosporum strain VKM F-3630D were studied. The range of exposure to the dose of the mutagen (incubation time and concentration of HMU) was determined. The lethal and sublethal doses, as well as the conditions under which the highest variability in the levels of CyA production was observed, were determined. It was shown that the most efficient method to further increase CyA production in the studied high-producer T. inflatum strain is mutagenic treatment with 0.1% HMU for 2 h. In this case, we observed a relatively high level of survival of clones (80–85%), a low level of their morphological variability (10–15%), and a wide range in the CyA production level (80–170%) as compared to the original strain. This enabled us to select a number of clones with an increased CyA production level. The yield of the target metabolite was maximal in clone no. 13-27, where the CyA production level increased by 1.6–1.7 times.



Fermentation of Detoxified Acid-Hydrolyzed Pyrolytic Anhydrosugars into Bioethanol with Saccharomyces cerevisiae 2.399
Abstract
Pyrolysate obtained from the pyrolysis of waste cotton is a source of fermentable sugars that could be fermented into bioethanol fuel and other chemicals via microbial fermentation. However, pyrolysate is a complex mixture of fermentable and non-fermentable substrates causing inhibition of the microbial growth. The aim of this study was to detoxify the hydrolysate and then ferment it into bio-ethanol fuel in shake flasks and fermenter applying yeast strain Saccharomyces cerevisiae 2.399. Pyrolysate was hydrolyzed to glucose with 0.2 M sulfuric acid, neutralized with Ba(OH)2 followed by treatment with ethyl acetate and activated carbon to remove fermentation inhibitors. The effect of various fermentation parameters such as inoculum concentration, pH and hydrolysate glucose was evaluated in shake flasks for optimum ethanol fermentation. With respect to inoculum concentration, 20% v/v inoculum i.e. 8.0 × 108–1.2 × 109 cells/mL was the optimum level for producing 8.62 ± 0.33 g/L ethanol at 9 h of fermentation with a maximum yield of 0.46 g ethanol/g glucose. The optimum pH for hydrolysate glucose fermentation was found to be 6.0 that produced 8.57 ± 0.66 g/L ethanol. Maximum ethanol concentration, 14.78 g/L was obtained for 4% hydrolysate glucose concentration after 16 h of fermentation. Scale-up studies in stirred fermenter produced much higher productivity (1.32 g/L/h–1) compared to shake flask fermentation (0.92 g/L/h–1). The yield of ethanol reached a maximum of 91% and 89% of the theoretical yield of ethanol in shake flasks and fermenter, respectively. The complex of integrated models of development was applied, that has been successfully tested previously for the mathematical analysis of the fermentation processes.



Study of the Protective Activity of Chitosan Hydrolyzate Against Septoria Leaf Blotch of Wheat and Brown Spot of Tobacco
Abstract
Chitosan hydrolyzate containing low-molecular-weight chitosan (≤24 kDa) and its oligomers (≤1.2 kDa) has been obtained via chemical depolimerization of high-molecular-weight chitosan by nitric acid. The fractions of the obtained chitosan hydrolyzate have been characterized by high performance gel permeation chromatography and proton magnetic resonance. The test performed on detached leaves of wheat has shown that the hydrolyzate completely inhibits the development of Stagonospora nodorum, a casual agent of the septoria leaf blotch, at a concentration of 200 μg/mL. A similar test with detached tobacco leaves has shown that the hydrolyzate at a concentration of 100 μg/mL also inhibits the development of Alternaria longipes, which causes brown spot of tobacco, by 75%.



Hydrolysis of Soybean Proteins with Kamchatka Crab Hepatopancreas Enzyme Complex
Abstract
To perform hydrolysis with the enzyme complex from the hepatopancreas of the Kamchatka crab, a protein mixture was isolated from soybean meal by extraction at alkaline pH values. Extractable low-molecular impurities were removed by ultrafiltration and precipitation of proteins with alcohol. The amino acid composition of the obtained protein extract turned out to be similar to the composition of the fish meal traditionally used in the production of fish feeds. Analysis of the products of fermentolysis by DDS-electrophoresis, HPLC, and mass spectrometry showed a high degree of hydrolysis of soybean proteins. Depending on the time of fermentolysis, the hydrolysates contained up to 60% (18 h of hydrolysis) of free amino acids (the fraction of the weight of the hydrolyzed protein mixture) and short peptides (2–20 amino acid residues).



Physicochemical Properties and Biological Activity of the Peptide–Protein Complex from Bovine Scleral Tissue
Abstract
The peptide–protein complex from bovine sclera was studied. It is shown that it contained a protein with a molecular weight of 66387 Da with the partial N-terminal amino acid sequence DTHKSEIAHRFKDLG-, which is homologous to the mature molecule of bovine serum albumin, and polypeptides with molecular weights of 1300–5080 Da. With a model of the organotypic cultivation of posterior eye tissues of the newt Pleurodeles waltl in vitro, it was shown that the effect of this peptide–protein complex in low doses increased the viability of scleral fibroblasts.



Effect of the Compositions of Sample and Polymer Sorbents on the Extraction of Volatile Compounds by Solid-Phase Microextraction
Abstract
A comparative study was performed by solid phase microextraction and capillary gas chromatography to establish the ability of four polymer sorbents of different compositions to extract and concentrate volatile organic compounds from the gas phase above an aqueous solution. All polymer sorbents sorbed nonpolar monoterpene hydrocarbons via a cooperative mechanism with almost equal and high efficiency. Sorbents based on polymethyl disiloxane and its mixture with divinylbenzene were more effective in extracting acetates and sesquiterpenes. As the concentration of these compounds in the gas phase increased, their binding by sorbents decreased. It was found that the determination of polar compounds depended on the presence of a solvent in the system. Compounds that are highly soluble in water (alcohols, ketones, etc.) had low coefficients of distribution between gas and water phases. Consequently, their sorption to any of the polymer sorbents was negligible. In the absence of the solvent, the degree of their extraction from the gas phase above the sample was high. It was shown that the actual composition of compounds in the initial mixture of essential oils could significantly differ from their composition in the gas phase. This method is convenient and informative for the purpose of studying the composition of volatile compounds in the gas phase that determine the flavor of the product.


