Exploration of GH94 Sequence Space for Enzyme Discovery Reveals a Novel Glucosylgalactose Phosphorylase Specificity

The substantial increase in DNA sequencing efforts has led to a rapid expansion of available sequences in glycoside hydrolase families. The ever-increasing sequence space presents considerable opportunities for the search for enzymes with novel functionalities. In this work, the sequence-function space of glycoside hydrolase family 94 (GH94) was explored in detail, using a combined approach of phylogenetic analysis and sequence similarity networks. The identification and experimental screening of unknown clusters led to the discovery of an enzyme from the soil bacterium Paenibacillus polymyxa that acts as a 4-O-β-d -glucosyl-d -galactose phosphorylase (GGalP), a specificity that has not been reported to date. Detailed characterization of GGalP revealed that its kinetic parameters were consistent with those of other known phosphorylases. Furthermore, the enzyme could be used for production of the rare disaccharides 4-O-β-d -glucosyl-d -galactose and 4-O-β-d -glucosyl-l -arabinose. Our current work highlights the power of rational sequence space exploration in the search for novel enzyme specificities, as well as the potential of phosphorylases for rare disaccharide synthesis.     

Synthesis and properties of YBa2Cu3O7-δ – Y2Ba4CuWO10.8 superconducting composites

YBa₂Cu₃O_7-δ (YBCO) is a well-known high-temperature superconductor. However, its critical current density and thus maximum trapped magnetic field can be improved significantly by introducing the secondary phases (artificial pinning centers). In this contribution, we successfully prepared YBCO single-grain bulks with Y₂Ba₄CuWO_10.8 phase serving as a source of pinning centers. This phase was prepared by solid-state reaction and further refined by milling. In the next step single-grain YBCO bulks with homogeneously distributed Y₂Ba₄CuWO_10.8 were prepared by top-seeded melt growth. Precursors as well as the final product were characterized by XRD, SEM and EDS. The phase composition of YBCO bulks containing Y₂Ba₄CuWO_10.8 was analyzed using Rietveld analysis. Thermal stability of YBCO bulk was studied by STA. Furthermore, PPMS was used to study electrical resistivity and critical current density. Bulk superconducting properties such as levitation force and trapped field ability were also measured.     

Extraction of Bitter Acids from Hops and Hop Products Using Pressurized Solvent Extraction (PSE)

The EBC method 7.7, currently used for determination of bitter acids in hop products, is a time‐consuming and laborious extraction technique. In this paper, our aim was to propose a new extraction method based on Pressurized Solvent Extraction (PSE) sometimes also called Pressurized Fluid Extraction (PFE) or Accelerated Solvent Extraction (ASE). Compared to conventional extractions, PSE offers a number of important benefits. PSE on OnePSE^® automated extractor was used for extraction of α‐ and β‐acids from hops and hop products and the parameters influencing extraction efficiency and the influence of the sample preparation method were studied. The quantitative determination of α‐ and β‐acids in the extracts was accomplished by using an HPLC apparatus equipped with diode array detector. The experimental results were compared with those obtained by the standard EBC 7.7 method and the two methods were found to be fully compatible     

Using Faster Gas Chromatography Analyses in Brewing Analytics

Dominant trends in analytical chemistry include miniaturization in sample preparation techniques and faster run times to provide high‐throughput screening, fast process monitoring and fast method development. This study focused on the application of narrow bore gas chromatographic capillary columns, 0.18 mm internal diameter, for brewing analyses. On these capillary columns, faster analyses could be performed compared to conventional GC capillary columns using 0.32 to 0.53 mm internal diameters. The robustness of the state‐of‐the‐art faster capillary gas chromatography, without compromising resolution, has been demonstrated with the analyses of beer flavour compounds such as lower and higher alcohols, esters and other volatile compounds such as acetaldehyde and dimethyl sulphide in beer. These methods were able to reduce sample run times by 60%.     

Structural Study of the Partially Disordered Full‐Length δ Subunit of RNA Polymerase from Bacillus subtilis

The partially disordered δ subunit of RNA polymerase was studied by various NMR techniques. The structure of the well‐folded N‐terminal domain was determined based on inter‐proton distances in NOESY spectra. The obtained structural model was compared to the previously determined structure of a truncated construct (lacking the C‐terminal domain). Only marginal differences were identified, thus indicating that the first structural model was not significantly compromised by the absence of the C‐terminal domain. Various ¹⁵N relaxation experiments were employed to describe the flexibility of both domains. The relaxation data revealed that the C‐terminal domain is more flexible, but its flexibility is not uniform. By using paramagnetic labels, transient contacts of the C‐terminal tail with the N‐terminal domain and with itself were identified. A propensity of the C‐terminal domain to form β‐type structures was obtained by chemical shift analysis. Comparison with the paramagnetic relaxation enhancement indicated a well‐balanced interplay of repulsive and attractive electrostatic interactions governing the conformational behavior of the C‐terminal domain. The results showed that the δ subunit consists of a well‐ordered N‐terminal domain and a flexible C‐terminal domain that exhibits a complex hierarchy of partial ordering.     

Integrated material flow and radioactivity distribution methodology within the calculation tool for evaluation of parameters for decommissioning of nuclear installations

Decommissioning of nuclear facilities becomes an important issue in all areas of nuclear technology, mainly in their energetic applications. Decommissioning process has to be planned in the safe, ecological and economic manner. It determines the requirements on appropriate evaluation of needed technologies, media, amount of solid materials released into the environment, radioactivity of effluents, amount of radioactive waste for disposal, number and exposure of personnel and finally the financial demands. A detailed evaluation of these parameters may be done by analytical calculation approach. This approach models a real process of decommissioning with its individual basic activities. The methodology of integrated material flow and radioactivity distribution within this calculation evaluation tool is applied and implemented to describe the real decommissioning activities and their mutual relations to obtain more accurate outputs.     

Antifungal activity of lauric acid derivatives against Aspergillus niger

The antifungal effects of lauric acid and four lauric acid derivatives (monolauroylglycerol, D-laurate A, T-laurate A, 6-O-lauroysucrose) were tested on the spore germination and the growth rate of Aspergillus niger DMF 0801. The results showed that the tested substances varied in their antifungal activity and they also confirmed the relation of the structure of tested substances and their antifungal effects. Monolauroylglycerol at concentration level between 0.2 and 1.8 mmol l^-1 caused the highest inhibition of spore outgrowths whereas 6-O-lauroysucrose at concentration between 0.05 and 1.8 mmol l^-1 did not exhibit inhibition of spore germination. The inhibition of colony growth rate was detected mainly at presence of 6-O-lauroysucrose.     

Acid-catalyzed decomposition of stable 1-(2,1-benzisothiazol-3-yl)-3-phenyltriazenes

The acid-catalyzed decomposition of unusually stable 1-(2,1-benzisothiazol-3-yl)-3-phenyltriazenes in either aqueous perchloric acid or an aqueous mixture of perchloric and acetic acid was studied under pseudo-first order reaction conditions at 25 °C. Different products were obtained according to substitution on nitrogen N-3. For a triazene carrying hydrogen, the corresponding 3-amino-2,1-benzisothiazole and benzenediazonium salts were formed whereas in the case of substitution by an alkyl group (methyl and n-butyl) the 2,1-benzisothiazole-3-diazonium salt and N-alkylaniline were obtained. The observed rate constant (k_obs) of the acid-catalyzed decomposition increased, initially, nonlinearly with increasing concentration of acid. Subsequently, k_obs decreased slightly and at high acid concentration, increased steeply once again. An A-S_E2 mechanism in which protonation of the triazene nitrogen proceeds simultaneously with cleavage of the N–N bond is proposed. Tautomerism of 1-(2,1-benzisothiazol-3-yl)-3-phenyltriazene was investigated using ¹H NMR spectroscopy.     

Determination of Heat of Adsorption on Clean Solid Surfaces

One of the basic parameters characterizing the bond between a chemisorbed atom (or molecule) and the surface is its dissociation energy. It is difficult, however, to determine the bond energy and therefore it becomes necessary to turn to some other quantity, which is sufficiently close to the bond energy and which can be determined without much difficulty. Such a quantity is the heat of adsorption. Heat of adsorption is defined as the difference between the total energy of the adsorption system prior to and after the adsorption process, i.e., it equals the sum of all energy changes connected with adsorption. Thus the experimentally determined value for the heat of adsorption is obviously closely related to the bond energy of the chemisorbed particle, though this relation is not quite simple [1,2].     

Emission Spectra of a Two-level Atom in a Kerr-like Medium

Abstract

We have investigated the spectrum of light emitted by a single atom interacting with a single mode of the radiation field in an ideal cavity filled with a Kerr-like medium. It is shown that owing to the Kerr-like nonlinearity in the system the spectrum of the emitted light exhibits a single-peaked structure for sufficiently high intensities of the initial coherent field instead of the triplet structure in the case of the standard Jaynes-Cummings model.