Effect of gelatin on properties of <Emphasis Type="Italic">Luciola mingrelica</Emphasis> firefly luciferase

An ATP-reagent containing thermostable mutant of Luciola mingrelica firefly luciferase, MgSO₄, components of buffer solution, and stabilizers, which is widely used to detect nano- and picomolar concentrations of adenosine-5′-triphosphate (ATP) in various biological samples, is been object of this study. The activity, stability, and analytical characteristics of the ATP-reagent have been assessed in the presence and absence of 5% gelatin and in gelatin gel. The solution of ATP-reagent was obtained at 30°C and a gelatin concentration of 5%, while gel formation occurred at 22°C. The gelatin addition decreased the activity and stability of luciferase slightly. The sensitivity of ATP detection (above 0.96) did not depend on the gelatin presence and the aggregate state of the disperse system. Limits of detection were 2 × 10⁻¹², 7 × 10⁻¹³, and 7 × 10⁻¹⁴ M ATP, when the ATP-reagent was used in gelatin films and in the solution in the presence of 5% gelatin and in the absence, respectively. It was shown that the storage of ATP-reagent in gelatin gel not only preserved enzymatic activity, but protected the enzyme from bacterial contamination, which was the cause of the enzyme activity loss.     

New Transformation of 1,3‐Oxathiolane‐2‐thione into 1,3‐Dithiolan‐2‐one via Polymerization and Depolymerization

This article deals with the cationic and anionic depolymerization of polydithiocarbonate, which was synthesized by cationic polymerization of 5‐phenoxymethyl‐1,3‐oxathiolane‐2‐thione ( 1 ) using methyl triflate as the initiator. The cationic depolymerization of the obtained polymer was carried out in the presence of 5–20 mol‐% of methyl triflate or triflic acid catalyst in chlorobenzene at 60 °C for 96 h to afford 4‐phenoxymethyl‐1,3‐dithiolan‐2‐one ( 2 ) in 35–83% yield. The anionic depolymerization of the polymer was carried out in the presence of 5 mol‐% of triethylamine or potassium tert‐butoxide at 20 °C for 24 h to afford 2 in 85–100% yield.     

Synthesis and properties of new aromatic polyamides with redox‐active 2,4‐dimethoxytriphenylamine moieties

A new triphenylamine‐based diamine monomer, 4,4′‐diamino‐2″,4″‐dimethoxytriphenylamine ( 2 ), was synthesized from readily available reagents and was reacted with various aromatic dicarboxylic acids to produce a series of aromatic polyamides ( 4a–h ) containing the redox‐active 2,4‐dimethoxy‐substituted triphenylamine (dimethoxyTPA) unit. All the resulting polyamides were readily soluble in polar organic solvents and could be solution cast into tough and flexible films. These polymers exhibited good thermal stability with glass transition temperatures of 243–289 °C and softening temperatures of 238–280 °C, 10% weight loss temperatures in excess of 470 °C in nitrogen, and char yields higher than 60% at 800 °C in nitrogen. The redox behaviors of the polymers were examined using cyclic voltammetry (CV). All these polyamides showed two reversible oxidation processes in the first CV scan. The polymers also displayed low ionization potentials as a result of their dimethoxyTPA moieties. In addition, the polymers displayed excellent stability of electrochromic characteristics with coloration change from a colorless neutral state to green and blue‐purple oxidized states. These anodically coloring polyamides showed high green coloration efficiency (CE = 329 cm²/C), high contrast of optical transmittance change (ΔT% = 84% at 829 nm), and long‐term redox reversibility. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 3392–3401, 2010     

Elastic properties and mechanical relaxation behaviors of PVDF (poly (vinylidene fluoride)) at temperatures between −20 and 100 °C and at 2 MHz ultrasonic frequency

The elastic properties of PVDF have been investigated as a function of temperature. The propagation velocity and absorption of longitudinal and transverse ultrasonic waves have been measured at a constant frequency of 2 MHz and temperatures between –20 and 100 °C. Hence, the temperature dependences of storage and loss elastic constants have been obtained for temperatures between –20 and 100 °C. It has been seen that the relaxation behavior is affected from the form of mechanical disturbance. For the longitudinal mode, only one relaxation peak at 42 °C, but for transverse mode three relaxation peaks at 28 °C, 60 °C, and 94 °C have been observed. The results have been compared with the literature values obtained previously for PVDF. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 2862–2873, 2005     

Ligand‐Free Copper‐Catalyzed Negishi Coupling of Alkyl‐, Aryl‐, and Alkynylzinc Reagents with Heteroaryl Iodides

Reported herein is an unprecedented ligand‐free copper‐catalyzed cross‐coupling of alkyl‐, aryl‐, and alkynylzinc reagents with heteroaryl iodides. The reaction proceeds at room temperature for the coupling of primary, secondary, and tertiary alkylzinc reagents with heteroaryl iodides without rearrangement. An elevated temperature (100 °C) is required for aryl–heteroaryl and alkynyl–heteroaryl couplings.     

Aromatic polyamides. III. Copoly(4,4′-oxanilideterephthalamide—4,4′-phenyleneterephthalamide): Synthesis,wet-spinning,fiber thermal characterization,and stability in sulfuric acid

Copoly(4,4′-oxanilideterephthalamide—4,4′-phenyleneterephthalamide) (A-202/PPD) was synthesized by reaction of 4,4′-diaminooxanilide, p-phenylenediamine, and terephthaloyl chloride in organic solvents. Copolymer inherent viscosities in H₂SO₄ as high as 10.3 were obtained. Isotropic copolymer solutions (4%—5% concentration) of A-202/40%–80% PPD were spun to fibers with tenacity/elongation/modulus at 1% extension in the 13–14 gpd/1.5%–2%/700–1000 gpd range. Oxamide and amide stabilities in 98–100% H₂SO₄ and 20% oleum were compared. Poly(4,4′-oxanilideterephthalamide) (A-202), A-202/PPD copolymers, and poly(4,4′-phenyleneterephthalamide) (PPT) were unstable in 20% oleum, but all proved relatively stable in 100% H₂SO₄. However, the oxamide linkage proved less stable than the amide linkage in 98% H₂SO₄. A-202 and A-202/PPD copolymers formed stable anisotropic spinning solutions in 1% oleum at 10–20% concentrations. Dynamic mechanical analyses (Vibron) showed no glass transition temperature (T_g) below 200°C. Dilatometric measurement of A-202/50% PPD revealed a T_g at 257°C. Differential thermal analyses of A-202/40–80% PPD exhibited endotherms at 470–480°C. Thermogravimetric analyses showed no significant weight loss below 400°C.     

(Thio)Amidoindoles and (Thio)Amidobenzimidazoles: An Investigation of Their Hydrogen‐Bonding and Organocatalytic Properties in the Ring‐Opening Polymerization of Lactide

The mechanism of the ring‐opening polymerization (ROP) of lactide catalyzed by two partner hydrogen‐bonding organocatalysts was explored. New amidoindoles 4 a , c , thioamidoindoles 4 b , d , amidobenzimidazoles 5 a , c , and thioamidobenzimidazoles 5 b , c were synthesized and used as activators of the monomer. In the solid state and in solution, compounds 4 and 5 showed a propensity for self‐association, which was evaluated. (Thio)Amides 4 and 5 do catalyze the ROP of lactide in the presence of a cocatalyst, tertiary amine 3 a or 3 b , which activates the growing polymer chain through hydrogen‐bonding. Reactions were conducted in 2–24 h at 20 °C; conversion yields ranged between 22 and 100 %. A detailed study of the intermolecular interactions undertaken between the participating species showed that, as expected, simultaneous weak hydrogen bonds do exist to activate the reagents. Moreover, interactions have been revealed between the partner catalysts 4 / 5 + 3 . ROP catalyzed by these partner activators is thus governed by multiple dynamic equilibria. The latter should be judiciously adjusted to fine‐tune the catalytic properties of (thio)amides and organocatalysts, more generally.     

Self-organization processes and synthesis of nanostructured silver microparticles in AgNO<Subscript>3</Subscript> solutions with variable pH values

The morphology of micrometer-sized silver particles obtained by liquid-phase chemical reduction of silver nitrate with ascorbic acid depends appreciably on the solution pH. The synthesis carried out at 100°C for 20 min at pH < 4 or pH > 9 yields anisotropic faceted nanocrystalline particles, while the synthesis at pH = 5–8 results in self-assembly to give microspheres representing close-packed aggregates of a huge number of silver nanoparticles with a cauliflower structure.     

Purification and Characterization of Hyaluronate Lyase from <Emphasis Type="Italic">Arthrobacter globiformis</Emphasis> A152

A hyaluronate lyase was obtained by cultivating Arthrobacter globiformis strain A152. The enzyme was purified to homogeneity from the supernatant by ammonium sulfate fractionation, Q Sepharose Fast Flow, and Sephadex G-100 chromatography. The purification resulted in a 32.78-fold increase in hyaluronate lyase activity with specific activity of 297.2 U/mg. The molecular weight of the enzyme determined by SDS-PAGE was approximately 73.7 kDa. Using hyaluronic acid (HA) as a substrate, the maximal reaction rate (V_max) and the Michaelis–Menten constant (K_m) of hyaluronate lyase were found to be 4.76 μmol/min/ml and 0.11 mg/ml, respectively. The optimum pH and temperature values for hyaluronate lyase activity were pH 6.0 and 42 °C, respectively. This enzyme was stable at pH 4–10, 5–7, and 5–7 at 4, 37, and 42 °C, respectively. Investigation about temperature effects on hyaluronate lyase displayed that it was stable at 30–37 °C and also showed high activity at 37 °C. The enzymatic activity was enhanced by Ca²⁺ and was strongly inhibited by Cu²⁺ and SDS. These properties suggested that the hyaluronate lyase in this study could bring promising prospects in medical and industry applications.     

Dielectric relaxation in ethylene–methacrylic acid copolymers and their salts

Dielectric relaxation data have been obtained for two ethylene–methacrylic acid copolymers (containing about 4 mole-% methacrylic acid units and about 8 mole-% methacrylic acid units, respectively) and the lithium, sodium, and calcium salts prepared by partial neutralization of the polyacids. The frequency range employed was from 50 Hz to 10 kHz and the temperature range was from ?130°C to 100°C. Attention is focused on three dielectric loss regions labeled β, β and α in order of increasing temperature. The β′ process (?10°C at 100 Hz in the salts only) correlates with a mechanical loss process previously reported and is attributed to microbrownian motion taking place in an amorphous hydrocarbon phase. The β′ process (20°C at 100 Hz) has also been observed mechanically and is attributed to the same mechanism as the β process. The higher temperature of this relaxation compared to the β relaxation is attributed to the presence of acid groups which form crosslinks composed of interchain hydrogen bonds. The α process (>50°C at 100 Hz in the salts only) correlates with dielectric and NMR data previously reported for a sodium salt and is assigned to motions within ionic domains formed by the clustering of salt groups.     

Certification by the Karl Fischer method of the water content in SRM 2890, Water Saturated 1-Octanol, and the analysis of associated interlaboratory bias in the measurement process

The calibration of Karl Fischer instruments and reagents and the compensation for instrumental bias are essential to the accurate measurement of trace levels of water in organic and inorganic chemicals. A stable, non-hygroscopic standard, Water Saturated Octanol, which is compatible with the Karl Fischer reagents, has been prepared. This material, Standard Reference Material (SRM) 2890, is homogeneous and is certified to contain 39.24 ± 0.85 mg water/mL (expanded uncertainty) of solution (47.3 ± 1.0 mg water/g solution, expanded uncertainty) at 21.5?°C. The solubility of water in 1-octanol has been shown to be nearly constant between 10?°C and 30?°C (i.e., within 1% of the value at 21.5?°C). The results of an interlaboratory comparison exercise illustrate the utility of SRM 2890 in assessing the accuracy and bias of Karl Fischer instruments and measurements.     

Continuous Flow Reduction of Artemisinic Acid Utilizing Multi‐Injection Strategies—Closing the Gap Towards a Fully Continuous Synthesis of Antimalarial Drugs

One of the rare alternative reagents for the reduction of carbon–carbon double bonds is diimide (HN?NH), which can be generated in situ from hydrazine hydrate (N₂H₄ ? H₂O) and O₂. Although this selective method is extremely clean and powerful, it is rarely used, as the rate‐determining oxidation of hydrazine in the absence of a catalyst is relatively slow using conventional batch protocols. A continuous high‐temperature/high‐pressure methodology dramatically enhances the initial oxidation step, at the same time allowing for a safe and scalable processing of the hazardous reaction mixture. Simple alkenes can be selectively reduced within 10–20 min at 100–120 °C and 20 bar O₂ pressure. The development of a multi‐injection reactor platform for the periodic addition of N₂H₄ ? H₂O enables the reduction of less reactive olefins even at lower reaction temperatures. This concept was utilized for the highly selective reduction of artemisinic acid to dihydroartemisinic acid, the precursor molecule for the semisynthesis of the antimalarial drug artemisinin. The industrially relevant reduction was achieved by using four consecutive liquid feeds (of N₂H₄ ? H₂O) and residence time units resulting in a highly selective reduction within approximately 40 min at 60 °C and 20 bar O₂ pressure, providing dihydroartemisinic acid in ≥93 % yield and ≥95 % selectivity.     

Iron‐catalyzed Cross‐Coupling of Propargyl Carboxylates and Grignard Reagents: Synthesis of Substituted Allenes

Presented herein is a mild, facile, and efficient iron‐catalyzed synthesis of substituted allenes from propargyl carboxylates and Grignard reagents. Only 1–5 mol % of the inexpensive and environmentally benign [Fe(acac)₃] at ?20 °C was sufficient to afford a broad range of substituted allenes in excellent yields. The method tolerates a variety of functional groups.     

Kinetic behavior of ethylene/1‐hexene copolymerization in slurry and solution reactors

The copolymerization of ethylene and 1‐hexene over a spherical polymer/MgCl₂‐supported TiCl₄ catalyst was studied as a function of the polymerization temperature from 40 to 100 °C in a slurry reactor and from 120 to 200 °C in a solution reactor with triethylaluminum (TEA) as a cocatalyst (1.0–6.8 mmol). The activities increased from 40 to 80 °C and then declined monotonically with increases in the temperature during the slurry and solution polymerizations. The kinetic behavior in the slurry and solution operations was described by the same rate expression. The modeling results indicated that the catalyst had at least two different types of catalytic sites; one site was responsible for the acceleration–decay nature of the activity profiles, whereas the second site resulted in long‐term activity. The apparent activation energy for site activation in the slurry operation was 69.9 kJ/mol; no activation energies for site activation could be estimated for the solution operation because the activation process was essentially instantaneous at the higher temperatures. The activation energies for deactivation were 100.3 kJ/mol for the slurry operation and 31.2 kJ/mol for the solution operation. The responses to TEA were similar for the slurry and solution operations; the rates increased with increasing amounts of TEA between 1.0 and 3.4 mmol and then decreased with larger amounts of TEA. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 2248–2257, 2005     

Bis(1,2-dimethoxyethan-O,O′)lithium-phosphanid, -arsanid und -chlorid – drei neue Vertreter des Bis(1,2-dimethoxyethan-O,O′)lithium-bromid-Typs

Metal Derivatives of Molecular Compounds. IX. Bis(1,2-dimethoxyethane- O,O′ )lithium Phosphanide, Arsanide, and Chloride – Three New Representatives of the Bis(1,2-dimethoxyethane- O,O′ )lithium Bromide Type Experiments to obtain thermally unstable lithium silylphosphanide at –60 °C from a 1,2-dimethoxyethane solution resulted in the isolation of its dismutation product bis(1,2-dimethoxyethane-O,O′)lithium phosphanide ( 1 ). The homologous arsanide 2 precipitated after a frozen solution of arsane in the same solvent had been treated with lithium n-butanide at –78 °C. Unexpectedly, too, the analogous chloride 3 and bromide 4 were formed in reactions of 1-chloro-2,2-bis(trimethylsilyl)-1λ³-phosphaethene with (1,2-dimethoxyethane-O,O′)lithium bis(trimethylsilyl)stibanide and of lithium 1,2,3,4,5-pentaphenyl-2,3-dihydro-1λ³-phosphol-3-ide with ω-bromostyrene, respectively. The monomeric complexes 1 {–100 ± 3 °C; a = 1391.1(4); b = 809.8(2); c = 1249.1(3) pm; β = 102.84(2)°}, 2 {–100 ± 3 °C; a = 1398.3(4); b = 819.8(3); c = 1258.5(4) pm; β = 103.35(2)°} and 3 {–100 ± 3 °C; a = 1308.4(2); b = 788.2(1); c = 1195.6(1) pm; β = 95.35(1)°} crystallize in the monoclinic space group C2/c with four solvated ion pairs in the unit cell; they are isotypic with bis(1,2-dimethoxyethane-O,O′)lithium bromide ( 4 ) {–73 ± 2 °C; a = 1319.0(2); b = 794.1(1); c = 1214.3(2) pm; β = 96.22(1)°}, already studied by Rogers et al. [13] at room temperature. The neutral complexes show a trigonal bipyramidal configuration of symmetry C₂, pnicogenanide or halide anions occupying equatorial sites {Li–P 260.4(4); Li–As 269.8(6); Li–Cl 238.6(7); Li–Br 256.3(10) pm} and the chelate ligands spanning equatorial and axial positions {Li–O_eq 205.4(4) to 207.4(4); Li–O_ax 208.9(3) to 215.5(2) pm}. The coordination within the (dme)₂Li fragment, the Li–X distances (X = P, As, Cl, Br), the structure of the chelate rings, and the packing of the neutral complexes are discussed in detail.     

Über Chalkogenolate. 175. Reaktion von Acetamidin mit Kohlenstoffdisulfid 4. Darstellung und Charakterisierung der N-Acetimidoyldithiocarbamidsäure

On Chalcogenolates. 175. Reaction of Acetamidine with Carbon Disulfide. 4. Synthesis and Characterization of N-Acetimidoyl Dithiocarbamic Acid Orange colored N-acetimidoyl dithiocarbamic acid has been prepared by reaction of the acetimidinium salt in aqueous solution with hydrochloric acid at 0°C. The properties and the thermal behaviour of the acid have been described, its decomposition in solution has been studied at 20°C kinetically.     

Synthesis and characterization of phenyl-pendant aromatic polythiazoles from bis-α-bromophenylacetyl compounds and dithioamides

Novel phenyl-pendant aromatic polythiazoles having inherent viscosities of 0.3–1.3 dL/g were synthesized by the solution polycondensation of bis[4-(α-bromophenylacetyl)phenyl] ether with aromatic dithioamides or dithiooxamide in dimethylformamide at 60°C. The polythiazole having m-phenylene linkage was readily soluble in chloroform and m-cresol, and transparent flexible film could be cast from the chloroform solution. Glass transition temperatures of these polythiazoles were in the range of 210–250°C. They started to decompose at about 500°C in air with 10% weight loss being recorded at around 570°C.     

The stress-related production of the activePhotinus pyralis and luciolaMingrelica firefly luciferases inEscherichia coli

The kinetics ofPhotinus pyralis andLuciola mingrelica luciferase gene expression was studied on plasmids with the thermoinducible λP_r promoter inEscherichia coli by SDS-gel electrophoresis of cell lysates to follow luciferase protein-synthesized, enzyme immunoassay (EIA) to follow native enzyme conformer, and the luciferase activity assay.E. coli cells were cultivated at temperature schemes 28–42–21°C or 28–21°C, or at alkali pH shift. In the cases of thermoinduction and pH shift, the luciferase expressions have similar features. The 3-h thermoinduction (42°C) followed by the incubation at 21°C, for 10 h resulted in the maximal amount of the luciferase protein of 4–5% of the total cell proteins. The yield did not change further. The amount of native luciferase conformer and the luciferase activity started to grow after incubation for 10 h at 21°C and reached the maximum after 50–60 h when the synthesized luciferase protein adopted the native-like conformation. At the same time, only 50% of the latter appeared to be catalytically active. An increase in the enzymatic activity correlates with an increase in the intracellular pH and ATP content. Intracellular metabolic reactions were shown to play a role in the conformational changes of the enzyme in a postthermoinduction period, and a possible mechanism of this effect is proposed.     

Cross‐linked Polymer‐Blend Gate Dielectrics through Thermal Click Chemistry

New cross‐linking reagents were synthesized and mixed with polystyrene (PS) in solution to form a blend. Thin‐films were spin‐coated from the blend and then cross‐linked by thermal activation at relatively low temperature (100 °C) to form cross‐linked gate dielectrics. This new method is compatible with plastic substrates in flexible electronics. The azide and alkyne cross‐linking reagents are kinetically stable at room temperature, so any premature cross‐linking is avoided during processing. This method also significantly improved the dielectric performances of PS thin films. Solution‐processed top‐gate organic field‐effect transistor devices with indacenodithiophene–benzothiadiazole copolymer as semiconductor layer and the cross‐linked PS blend as dielectric layer showed improved performances with lower gate leakages and higher operation stabilities than devices with neat PS film as dielectric layer.     

Co oxidation with oxygen in the presence of hydrogen on CoO/CeO<Subscript>2</Subscript> and CuO/CoO/CeO<Subscript>2</Subscript> catalysts

The catalytic activity of the CoO/CeO₂ and CuO/CoO/CeO₂ systems in selective CO oxidation in the presence of hydrogen at 20–450°C ([CuO] = 1.0–2.5%, [CoO] = 1.0–7.0%) is reported. The maximum CO conversion (X) decreases in the following order: CuO/CoO/CeO₂ (X = 98–99%, T = 140–170°C) > CoO/CeO₂ (X = 67–84%, T = 230–240°C) > CeO₂ (X = 34%, T = 350°C). TPD, TPR, and EPR experiments have demonstrated that the high activity of CuO/CoO/CeO₂ is due to the strong interaction of the supported copper and cobalt oxides with cerium dioxide, which yields Cu-Co-Ce-O clusters on the surface. The carbonyl group in the complexes Co^δ+-CO and Cu⁺-CO is oxidized by oxygen of the Cu-Co-Ce-O clusters at 140–160°C and by oxygen of the Co-Ce-O clusters at 240°C. The decrease in the activity of the catalysts at high temperatures is due to the fact that hydrogen reduces the clusters on which CO oxidation takes place, yielding Co⁰ and Cu⁰ particles, which are inactive in CO oxidation. The hydrogenation of CO into methane at high temperatures is due to the appearance of Co⁰ particles in the catalysts.