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A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3.
From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 °C. Specific activity of StmPr2 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 17 ± 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2.
This study describes a label-free impedimetric sensor based on short ssDNA recognition elements for the detection of hybridization events. We concentrate on the elucidation of the influence of target length and recognition sequence position on the sensorial performance. The impedimetric measurements are performed in the presence of the redox system ferri-/ferrocyanide and show an increase in charge transfer resistance upon hybridization of ssDNA to the sensor surface. Investigations on the impedimetric signal stability demonstrate a clear influence of the buffers used during the sensor preparation and the choice of the passivating mercaptoalcanol compound. A stable sensor system has been developed, enabling a reproducible detection of 25mer target DNA in the low nanomolar range. After hybridization, a sensor regeneration can be reached with deionized water by adjustment of effective convection conditions, ensuring a sensor reusability. By investigations of longer targets with overhangs exposed to the solution, we can demonstrate applicability of the impedimetric detection for longer ssDNA. However, a decreasing charge transfer resistance change (ΔRct) is found by extending the overhang. As a strategy to increase the impedance change for longer target strands, the position of the recognition sequence can be designed in a way that a small overhang is exposed to the electrode surface. This is found to result in an increase in the relative Rct change. These results suggest that DNA and consequently negative charge near the electrode possess a larger impact on the impedimetric signal than DNA further away.
One of the priority trends of carbon nanotechnology is creation of nanocomposite systems. Such carbon nanostructured composites were produced using - raw materials based on the products of agricultural waste, such as grape stones, apricot stones, rice husk. These products have a - wide spectrum of application and can be obtained in large quantities. The Institute of Combustion Problems has carried out the work on synthesis of the nanostructured carbon sorbents for multiple applications including the field of biomedicine. The article presents the data on the synthesis and physico-chemical properties of carbonaceous sorbents using physicochemical methods of investigation: separation and purification of biomolecules; isolation of phytohormone - fusicoccin; adsorbent INGO-1 in the form of an adsorption column for blood detoxification, oral (entero) sorbent - INGO-2; the study of efferent and probiotic properties and sorption activity in regard to the lipopolysaccharide (LPS), new biocomposites - based on carbonized rice husk (CRH) and cellular microorganisms; the use of CRH in wound treatment. A new material for blood detoxication (INGO-1) has been obtained. Adsorption of p-cresyl sulfate and indoxyl sulfate has shown that active carbon adsorbent can remove clinically significant level of p-cresyl sulfate and indoxyl sulfate from human plasma. Enterosorbent INGO-2 possesses high adsorption activity in relation to Gram-negative bacteria and their endotoxins. INGO-2 slows down the growth of conditionally pathogenic microorganisms, without having a negative effect on bifido and lactobacteria. The use of enterosorbent INGO-2 for sorption therapy may provide a solution to a complex problem - detoxication of the digestive tract and normalization of the intestinal micro ecology. The immobilized probiotic called "Riso-lact" was registered at the Ministry of Health of the Republic of Kazakhstan as a biologically active food additive. The developed technology is patented and provides production of the medicine in the form of freeze-dried biomass immobilized in vials.
Engineering of functional interfaces / Patrick Wagner ; Theodor Doll ; Michael J. Schöning (eds.)
(2014)
A novel photoexcitation method for the light-addressable potentiometric sensor (LAPS) is proposed to achieve a higher spatial resolution of chemical images. The proposed method employs a combined light source that consists of a modulated light probe, which generates the alternating photocurrent signal, and a ring of constant illumination surrounding it. The constant illumination generates a sheath of carriers with increased concentration which suppresses the spread of photocarriers by enhanced recombination. A device simulation was carried out to verify the effect of constant illumination on the spatial resolution, which demonstrated that a higher spatial resolution can be obtained.
Heat production in the windings of the stators of electric machines under stationary condition
(2014)
In electric machines due to high currents and resistive losses (joule heating) heat is produced. To avoid damages by overheating the design of effective cooling systems is required. Therefore the knowledge of heat sources and heat transfer processes is necessary. The purpose of this paper is to illustrate a good and effective calculation method for the temperature analysis based on homogenization techniques. These methods have been applied for the stator windings in a slot of an electric machine consisting of copper wires and resin. The key quantity here is an effective thermal conductivity, which characterizes the heterogeneous wire resin-arrangement inside the stator slot. To illustrate the applicability of the method, the analysis of a simplified, homogenized model is compared with the detailed analysis of temperature behavior inside a slot of an electric machine according to the heat generation. We considered here only the stationary situation. The achieved numerical results are accurate and show that the applied homogenization technique works in practice. Finally the results of simulations for the two cases, the original model of the slot and the homogenized model chosen for the slot (unit cell), are compared to experimental results.
Differentiation between Phaeocystis pouchetii (Har.) Lagerheim and Phaeocystis globosa Scherffel
(1987)
Molecular-genetic identification of emerged novel invasive pathogens of Asiatic Elm Ulmus pumila L
(2014)
The dwarf elm Ulmus pumila L. (Ulmaceae) is one of indigenous species of flora in Kazakhstan and forms a basis of dendroflora in virtually all settlements of the region. In the past decade, multiple outbreaks of previously unknown diseases of the small-leaved elm have been registered. In our study, by the molecular-genetic analysis it was found that the pathogens responsible for the outbreaks are microfungi belonging to the genus Fusarium – F. solani and F. oxysporum. The nucleotide sequences (ITS regions) isolated from the diseased trees showed very high similarity with the GenBank control numbers EU625403.1 and FJ478128.1 (100.0 and 99.0 % respectively). Oncoming research will focus on the search of natural microbial antagonists of the discovered phytopathogens.