Electrochemical determination of estradiol using a poly(l-serine) film-modified electrode

A thin film of poly(l-serine) was prepared via electropolymerization for the determination of trace levels of estradiol. In pH 5.0 phosphate buffer, l-serine was oxidized during the cyclic potential sweeps between −0.60 and 2.0 V, forming a thin film at the electrode surface. The electrochemical behavior of estradiol was investigated. The oxidation peak potential of estradiol shifts negatively at the poly(l-serine) film-coated glassy carbon electrode (GCE) compared with that at the bare GCE. Otherwise, the oxidation peak current greatly increases at the poly(l-serine) film-modified GCE. These phenomena suggest that the poly(l-serine) film exhibits catalytic activity towards the electrochemical oxidation of estradiol. Based on this, a sensitive, rapid and simple electrochemical method was proposed for the determination of estradiol. The limit of detection is evaluated to be 2.0 × 10⁻⁸ mol L⁻¹. Finally, this method was successfully used to determine estradiol in blood serum.     

The total synthesis of dilinoleoylphosphatidylserine and its activity in blood clotting systems

A total synthesis ofdl-phosphatidyl-(dilinoleoyl)-l-serine was achieved by the acylation of the barium salt of the phthalimidomethyl ester of glycerophosphoryl-n-anisyloxycarbonyl-l-serine. The dilinoleoyl intermediate was treated with hydrazine to remove the phthalimidomethyl group and with hydrogen chloride to remove the anisyloxycarbonyl protecting group. The resulting phosphatidylserine was purified by Rouser's methods, solubilized, and tested for biological activity in the antithromboplastin, recalcification, and Hicks-Pitney tests. It was found to have about the same anticoagulant activity as beef brain phosphatidylserine and hence was more active than the less unsaturated phosphatidylserine synthesized earlier. An abstract of the material in this paper appeared in Fed. Proc.24, No. 727 (1965).     

Surface active N-acylglutamate: III. Physicochemical properties of sodium long chain N-acylglutamates

The physicochemical properties of sodium long chain N-acylglutamate (AGS_n) are described. The wetting power, emulsifying power, dispersing power and calcium ion stability were measured. Monosodium N-lauroyl-l-glutamate (l-LGS) had good wetting and emulsifying properties, but disodium N-acylglutamate (AGS₂) was inferior tol-LGS in those properties. In the dispersing power using carbon black as powder,l-LGS was inferior but AGS₂, having a palmitoyl or stearoyl radical, was excellent. The AGS_n with a few exceptions had good calcium ion stabilities.     

Chemo-enzymatic synthesis of amino acid-based surfactants

The application of lipases to the synthesis of amino acid-based surfactants was investigated. Low yields (2–9%) were obtained in the acylation of free amino acids, such as l-serine and l-lysine, as well as their ethyl esters and amides with fatty acids, owing in part to low miscibility of the reactants. When the N-carbobenzyloxy (Cbz)-l-amino acids were used in an effort to improve miscibility of the amino acid derivatives with the acyl donor, a dramatic improvement was observed for N-Cbz-l-serine (92% yield) but not for N _α-Cbz- or N _ζ-Cbz-l-lysine (7 and 2% yield, respectively). As an alternative, and efficient synthesis of N _ζ-acyl-l-lysines was developed, based on the regiospecific chemical acylation of copper(II) lysinate. In pursuit of a general route to amino acid-fatty acid surfactants, the utility of a polyol linker was investigated. Thus, the glycerol ester of N _α′ N _ζ-di-Cbz-l-lysine was prepared and evaluated as a substrate for acylation. As expected, this and other glycer-1-yl esters of N-protected amino acids were excellent substrates for lipase-catalyzed acylation. Their reaction with myristic acid in the presence of Novozyme resulted in the regioselective acylation of the primary hydroxyl group of the glycerol moiety to afford the corresponding 1-O-(N-Cbz-l-aminoacyl)-3-O-myris-toylglycerols with conversions of 50–90%. These were readily deprotected to give a range of 1-O-(aminoacyl)-3-O-myristoyl-glycerols with overall yields of 27–71%.     

Continuous production of acyl <Emphasis Type="SmallCaps">l</Emphasis>-ascorbates using a packed-bed reactor with immobilized lipase

Saturated acyl (6-O-caproyl, lauroyl, and myristoyl) and unsaturated acyl (6-O-oleoyl, linoleoyl, and arachidonoyl) l-ascorbates were continuously synthesized at 50°C using a system where a column packed with ascorbic acid powder and a packed-bed reactor with an immobilized lipase from Candida antarctica were connected in series. A productivity of 1.6–1.9 kg/L reactor·d was achieved for at least 11 d. The surface tension of the caproyl or lauroyl l-ascorbate in aqueous solution was measured at various temperatures and pH to estimate the critical micelle concentration (CMC) of the acyl l-ascorbate. The CMC values were independent of temperature but dependent on the pH. The value of the caproyl ascorbate increased with an increase in pH.     

Enzymatic method of increasing phosphatidylcholine content of lecithin

An enzymatic method was established to increase the phosphatidylcholine (PC) contents of soybean and egg lecithins. Other phospholipids of lecithin were phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidic acid (PA). Seven preparations of phospholipase D (PLD), PLD-1 to PLD-6 ofStreptomyces origin and PLD-7 of cabbage origin, were tested for their ability to increase PC by transphosphatidylation in the presence of choline chloride (CC). The reactions were carried out at 30 C in a biphasic system that consisted of an aqueous phase containing PLD along with a buffer (optimum pH) having desired concentration of CC and Ca²⁺ and an ethyl acetate phase containing lecithin phospholipids. Intermitttent samples were extracted and analyzed by HPLC. Four of six PLD’s ofStreptomyces origin showed good transphosphatidylation (increase of PC contents of soybean lecithin from approximately 35% to 60–70% on a phospholipid basis) at 2.5 M CC, but the other two microbial PLD’s completely hydrolyzed the phospholipids to PA. Cabbage PLD-7 showed poor transphosphatidylation. PLD-3 gave the highest PC contents (70%) at 1.75 M CC. One hundred percent transphosphatidylation of pure PE to PC was achieved with PLD-3. PI was inert to the attack of most PLD preparations examined with the exception that PLD-3 hydrolyzed PI significantly. Purified PI could not be transphosphatidylized to PC; 100% PA was formed. Soybean lecithin containing about 80% PC and purified egg yolk lecithin with 75% PC could be converted to products having 95% PC and almost 100% PC, respectively, by PLD-3 at 1.75M CC. Studies on Enzymatic Conversion of Phospholipids (v)     

Preparation of calcium sulfate dihydrate and calcium sulfate hemihydrate with controllable crystal morphology by using ethanol additive

Calcium sulfate hemihydrate (CSH) with controlled crystal morphology has attracted broad interests due to its superior physical and chemical properties, as well as excellent biological performance. In this study, calcium sulfate dehydrate (CSD) was firstly synthesized via the reaction of H₂SO₄ and Ca(OH)₂ using ethanol as morphology modifier. The prepared CSD was then converted to CSH through a hydrothermal method. It was found that the precipitation time of CSD powders was dramatically shortened and the morphology of CSD crystals was changed from thick tabular to short-rod with the increment in ethanol addition. The finally-obtained CSH crystals were found to have hexagonal prisms shape with smaller aspect ratios. The CSH powder with the desired crystal morphology would provide improved setting behavior and biological performance of the CSH bone cement.     

Surface active N-acylglutamate: V. Application of N-acylglutamates to detergent bars

An application of sodium N-acyl-l-glutamate for detergent bars is described. The acyl radicals of cocoyl, hydrogenated tallowyl, semihydrogenated tallowyl, and lauroyl were used in the glutamates. The powder of sodium N-acyl-l-glutamate was mixed with a suitable amount of water and milled; then the mixture was extruded and stamped into a bar. Bars were examined relating composition of acyl radicals of sodium N-acyl-l-glutamates and properties of the bars or milling and molding properties. The other characteristics of the detergent bars were examined. It was found that attractive detergent bars can be made by using monosodium N-acyl-l-glutamate as a component; that hardness, solubility, and lather characteristics of bars can be controlled by blending various monosodium N-acyl-l-glutamates and adding moisture; that adjuvant can be used to improve the milling and stamping properties and characteristics of the monosodium N-acyl-l-glutamate bar; and that the bars are less irritating and leave the skin feeling good.     

Enzymatic synthesis of l-menthyl esters in organic solvent-free system

l-Menthol has been widely used as a food additive and an ingredient of cosmetics, and it is esterified to moderate the strong flavor. We attempted esterification of l-menthol with long-chain unsaturated fatty acid in an organic solvent-free enzymatic system. Commercially available lipases were screened, and Candida rugosa lipase was selected as a catalyst. Several factors affecting the esterification were investigated, and the reaction conditions were determined as follows: A reaction mixture of l-menthol/fatty acid (1:3, mol/mol), 30% water, and 700 units of the lipase per gram of reaction mixture was incubated at 30°C with stirring. After 24 h under these conditions, the esterification extents of l-menthol with oleic, linoleic, and α-linolenic acids reached 96, 88, and 95%, respectively. The structure of the esterified product was confirmed by mass, infrared, and nuclear magnetic resonance spectroscopies. Bacause Candida lipase acted strongly on l-menthol and very weakly on d-menthol, dl-menthol was esterified with oleic acid under the same conditions. The reaction showed high enantioselectivity; the enantiomeric ratio (E) was 31, and enantiomeric excess (ee) of l-menthyl oleate reached 88% after 32 h.     

A Novel Photocatalytic Conversion of Tryptophan to Kynurenine Using Black Light as a Light Source

Abstract  

The photocatalytic conversion of an aqueous solution of l-tryptophan (Trp) to kynurenine (KN) was investigated under the illumination of different light sources. Results show that Trp converted to KN with a selectivity of 64% under the illumination of a medium pressure (MP) Hg lamp. KN selectivity was increased to >90% when black light (BL) was used a light source. The novel use of BL in the photocatalytic conversion of Trp to KN significantly reduces the energy consumption compared with MP light.     

Olive oil microemulsions as a biomimetic medium for enzymatic studies: Oxidation of oleuropein

Microemulsions consisting of olive oil as the non-polar solvent, lecithin as surfactant, 1-propanol as cosurfactant, and water were prepared. The choice of the compositions of the microemulsions used was based on the pseudo-ternary phase diagrams of the system determined at 30°C, for different weight ratios of lecithin/olive oil. Lecithin solubilization and water incorporation in these microemulsion systems was limited. Tyrosinase, an oxidizing enzyme present in olives, was successfully incorporated in the water core of these microemulsions. Enzymatic oxidation of oleuropein, the most abundant olive phenolic compound, in the restricted aqueous environment of olive oil microemulsions was studied. Formation of oleuropein oxidation products was followed spectrophotometrically at 30°C for several minutes. An absorption maximum was observed at 415 nm. When the enzymatic reaction was considered at different tyrosinase and oleuropein concentrations, a rapid inactivation of the enzyme was observed. Addition of l-proline as a coupling reactor did not succeed in preventing enzyme inactivation in the microemulsions, probably owing to substrate localization and product accumulation around the entrapped enzyme molecules in the micellar interface.     

Purification of l-α-Glycerylphosphorylcholine from the Enzyme Reaction Solutions by Resin Column Chromatography

Colorless l-α-glycerylphosphorylcholine (l-α-GPC) was obtained at 98.8% purity, and with a specific rotation of −2.5° via a four-step procedure. l-α-GPC was first produced by phospholipase A₁ hydrolysis of phosphatidylcholine in an aqueous medium. Almost all the impurities were then removed using two successive D001 cation and D301-111 anion exchange resin column chromatography procedures, followed by decoloration with active carbon to remove minor pigments and remaining impurities. D001 resin and D301-111 resin showed remarkable ability for l-α-GPC isolation after being used eight times. Thus, this study presents a simple and cost-effective method for preparing l-α-GPC with high purity, low costs, and environmental friendliness, and encourages future investigation into its adaptation for industrial applications.     

Optimization of parameters through Taguchi method for enhanced corrosion inhibition of poly (vinyl alcohol-serine)

Parametric optimization was carried out for the mixture of polyvinyl alcohol (PVA) and l-serine using Taguchi statistical approach. L₉ orthogonal array of experiments were designed and the results were analyzed with Qualitek software. Synergistic effect of the mixture was enhanced at the Qualitek predicted optimized conditions. The graft polymer, poly(vinyl alcohol-serine) (PVAS), was synthesized using free radical-initiated reactions of l-serine with PVA in order to achieve effective inhibition. PVAS was characterized using Fourier transform infrared spectroscopy, scanning electron microscopy, and powder X-ray diffraction analysis. The thermal properties of PVAS were studied by thermogravimetry, differential thermal analysis, and differential scanning calorimetry. The corrosion inhibition property of PVAS on mild steel corrosion was continuously monitored using electrochemical techniques. Temperature effect on the inhibition properties of PVAS was studied using electrochemical and gravimetric measurements. AFM studies confirmed the formation of polymeric film on mild steel surface which inhibits the rate of corrosion. Theoretical calculations using Hyperchem 7.0 proved the better corrosion mitigation properties of PVAS compared to PVA. The experimental results and surface analysis confirmed the effective inhibition process of the graft polymer which was also supported by the theoretical calculations. A good correlation was obtained between the results of the electrochemical measurements, conventional gravimetric measurement, surface analysis, and theoretical calculations. All the studies proved the enhanced corrosion inhibition properties of PVA through grafting of polyserine.     

A new approach to the synthesis of nanosized powder CaO and its application as precursor for the synthesis of calcium borates

A procedure for the synthesis of calcium oxide has been developed, which consists in heat treatment of an aqueous solution of calcium acetate and d-glucose at 350 and then at 700 °C. The process parameters have been determined. It has been shown that when d-glucose is used and the reaction mixture is heat treated at 700 °C, highly dispersed calcium oxide with an average particle size of 77 nm is formed. CaO formed has been used as a precursor for the synthesis of priceite (Ca₂(B₅O₇)(OH)₅ · H₂O) during hydrothermal treatment of CaO in an aqueous solution of boric acid. It has been found that the optimal conditions for the synthesis of monophase priceite under hydrothermal conditions include the temperature range of 170–200 °C and the heating time of 12 h. When heating priceite at 800 °C in air for 1 h, highly dispersed powder calcium bis(borate) Ca(BO₂)₂ has been isolated as the final product. It is shown that the use of synthesized powder CaO leads to the formation of fine-crystalline powders of calcium borates. Samples obtained at each stage of the proposed synthesis of highly dispersed calcium oxide and calcium borates, using the example of priceite and calcium bis(borate), have been studied by powder X-ray diffraction, SEM, TEM, BET, IR spectroscopy, and DTA.     

Ceramic matrix composites in the alumina/5-30 vol.% YAG system

The preparation technique of the particulate composite materials in the alumina/YAG system was elaborated. Within alumina particles suspension yttria precursor was precipitated with ammonium carbonate. Drying and calcination at 600 °C resulted in the mixture of alumina and yttria particles, the latter being much finer than alumina particles. This mixture was additionally homogenized by short attrition milling in an aqueous suspension. Sintering of such powders results in the materials composed of YAG inclusions of sizes smaller than shown by alumina grains and evenly distributed within the matrix. YAG particles result from the reaction of Y₂O₃ with Al₂O₃ during heat treatment. YAG inclusions limit effectively grain growth of the alumina matrix. Hardness, fracture toughness, strength, Young modulus and wear susceptibility of composites and pure alumina were measured. Composites show higher hardness and in some cases higher fracture toughness and wear resistance than pure alumina polycrystals.     

Mechanochemical Synthesis of Hydroxyapatite from Calcium Oxide and Brushite

Fine hydroxyapatite (HA) powders were prepared by mechanically activating a mixture of calcium oxide and brushite powders in a high-energy shaker mill. A defective HA phase was formed when the starting powder mixture was activated for ≤20 h. When the mixture was calcined at 800°C, it was converted to β-tricalcium phosphate. In contrast, a nanocrystalline HA phase was formed when the mechanical activation was extended to 30 h. The material was transformed to a HA compound (Ca₁₀(PO₄)₆(OH)₂) of high crystallinity when it was calcined at 800°C.     

Influence of pH and Ionic Impurities on the Adsorption of Poly(acrylic) Dispersant onto a Zinc Oxide Surface

We have studied the properties of aqueous suspensions of ZnO powders with different purities. Our results suggest that powder purity determines the amount of dispersant necessary to form a stable aqueous suspension as well as the maximum adsorption capacity of the dispersant: the higher the positive surface charge of the as-received ZnO powders, the higher the amount of dispersant adsorbed onto the metal oxide surface. The surface charge of the ZnO particles in suspension is affected by the concentration of zinc as well as sulfate ions, which are the major impurities in the supernatant. The pH of the aqueous ZnO suspensions increases with increasing concentration of poly(acrylic) dispersant until the maximum adsorption capacity is attained. Further additions of dispersant do not increase the pH because of a buffer formation with impurity ions.     

A novel ∈-lysine acylase from <Emphasis Type="Italic">Streptomyces mobaraensis</Emphasis> for synthesis of <Emphasis Type="Italic">N∈</Emphasis>-acyl-<Emphasis Type="SmallCaps">l</Emphasis>-lysines

A novel ∈-lysine acylase (N ₆-acyl-l-lysine amidohydrolase; EC 3.5.1.17) was isolated from Streptomyces mobaraensis and purified to homogeneity by SDS-PAGE from the culture broth. The purified enzyme was monomeric, with a molecular mass of approximately 60 kDa. The enzyme was inactivated by the presence of 1,10-phenanthroline and activated in the presence of Co²⁺ and Zn²⁺. The enzyme showed a pH optimum of 8.0 and was stable at temperatures of up to 50°C for 1 h at pH 8.0. The enzyme specifically catalyzed the hydrolysis of the amide bond of various N∈-acyl-l-lysines. Furthermore, the enzyme efficiently catalyzed the synthesis of N∈-acyl-l-lysines with fatty and aromatic acyl groups in an aqueous buffer. In the syntheses of N∈-decanoyl-l-lysine, N∈-lauroyl-l-lysine, and N∈-myristoyl-l-lysine, the product precipitated and the yield was 90% or higher using 10 mM FA and 0.5 M l-lysine as the substrate.     

Kinetics of formation of fluorescent products from hexanal andl-lysine in a two-phase system

Kinetics of formation of fluorescent condensation products from hexanal andl-lysine (or itsN-acetylated forms) including mass-transfer has been studied in a two-phase system consisting of lysine (or lysine derivative) in a aqueous phosphate buffer and a 1-octanol solution of hexanal as model for formation of fluorophores between protein and carbonyl compounds in peroxidizing biological systems. The initial rate of formation of fluorescent products in the aqueous phase was found to be proportional to the concentration of hexanal and lysine and to increase in both phases with increasing pH in the aqueous phase, in contrast to a higher-order dependence on hexanal in the octanol phase. At pH=6.8, the temperature dependence of the appearance of fluorescent products corresponds to apparent energies of activation of 63 kJ·mol⁻¹ and 87 kj·mol⁻¹ in the aqueous phase and the octanol phase, respectively. Fluorescent condensation products appeared faster in the octanol phase. However, by a kinetic analysis, the fluorescent products were shown to be formed in the aqueous phase, corresponding to the lower energy of activation and to the simple second-order kinetics, and subsequently distributed between the aqueous phase and the octanol phase.l-Lysine reacted faster thanN _α-acetyl-l-lysine which reacted faster thanN _ε-acetyl-l-lysine. Using fluorescence quantum yields, determined to be 1.4·10⁻² in octanol and 8·10⁻³ in water at pH 6.8, an apparent partition coefficient of 17 (octanol/water) was determined for the condensation product ofl-lysine. The steady-state fluorescence in the octanol phase was attributed to two components with fluorescence life-time at 25°C of 0.7±0.05 ns and 5.1±0.2 ns, assigned to hexanal and the condensation product, respectively. The emission spectra, were resolved in the two components using phase-sensitive detection, and the condensation product had emission maximum at 405 nm.     

Enzymatic synthesis of N-acyl-l-amino acids in a glycerol-water system using acylase I from pig kidney

N-Medium- and long-chain acyl-l-amino acids were enzymatically synthesized from the corresponding l-amino acids and fatty acids using a reverse hydrolysis. Enzymes that are suitable for the synthetic reaction of N-acyl-l-amino acids were screened on the basis of hydrolytic activity toward N-lauroyl-l-glutamic acid as an indicator. Acylase I from pig kidney (EC 3.5.1.14) showed the highest N-acyl-l-amino acid hydrolytic activity among 57 commercially available enzymes tested. Acylase I effectively catalyzed the synthesis of N-lauroyl-l-amino acids except for N-lauroyl-l-proline and N-lauroyl-l-tyrosine in a glycerol-water system. Under the optimized reaction conditions, N-lauroyl-l-arginine and N-lauroyl-l-glutamic acid were obtained in conversions of 82 and 44%, respectively. The equilibrium constants calculated from the conversion obtained were 5.6, 15.4, 18.0, and 39.4 for the syntheses of N-lauroyl-l-glutamic acid, N_α-lauroyl-l-lysine, N-lauroyl-l-glutamine, and N-lauroyl-l-methionine, respectively. N-Acyl-l-arginines with myristic acid and palmitic acid as the fatty acid were also synthesized using acylase I.