sodium-dodecyl-sulfate and imidazole

A novel peroxidase-like artificial enzyme, named "caseoperoxidase", was biomimetically designed using a nano artificial amino acid apo-protein hydrophobic pocket. This four-component nano artificial enzyme containing heme-imidazole-β-casein-SDS exhibited high activity growth and k(cat) performance toward the native horseradish peroxidase demonstrated by the steady state kinetics using UV-vis spectrophotometry. The hydrophobicity and secondary structure of the caseoperoxidase were studied by ANS fluorescence and circular dichroism spectroscopy. Camel β-casein (Cβ-casein) was selected as an appropriate apo-protein for the heme active site because of its innate flexibility and exalted hydrophobicity. This selection was confirmed by homology modeling method. Heme docking into the newly obtained Cβ-casein structure indicated one heme was mainly incorporated with Cβ-casein. The presence of a main electrostatic site for the active site in the Cβ-casein was also confirmed by experimental methods through Wyman binding potential and isothermal titration calorimetry. The existence of Cβ-casein protein in this biocatalyst lowered the suicide inactivation and provided a suitable protective role for the heme active-site. Additional experiments confirmed the retention of caseoperoxidase structure and function as an artificial enzyme.

Topics: Binding Sites; Biocatalysis; Biomimetics; Caseins; Catalytic Domain; Circular Dichroism; Hemin; Horseradish Peroxidase; Hydrophobic and Hydrophilic Interactions; Imidazoles; Kinetics; Models, Molecular; Multiprotein Complexes; Nanoparticles; Protein Binding; Protein Conformation; Protein Structure, Secondary; Sodium Dodecyl Sulfate; Spectrophotometry

Environmental remediation is a highly pressing issue in society. Here we demonstrate that autonomous self-propelled millimeter sized capsules can sense the presence of pollutants, mark sites for visible identification and remove the contamination, while navigating in a complex environment of interconnected channels, the maze. Such long-range self-powered capsules propelled by the Marangoni effect are capable of releasing chemicals to alter the pH and induce aggregation during pollutant flocculation at a faster rate than convection or diffusion. These devices are foreseen to have real-world environmental applications in the near future.

Topics: Capsules; Chemical Precipitation; Dimethylformamide; Energy Transfer; Environmental Monitoring; Environmental Restoration and Remediation; Equipment Design; Flocculation; Hydrogen-Ion Concentration; Imidazoles; Materials Testing; Metal Nanoparticles; Microfluidic Analytical Techniques; Nanocapsules; Polymers; Printing, Three-Dimensional; Sodium Dodecyl Sulfate; Sulfones; Surface-Active Agents; Video Recording; Water Pollutants

A new cationic dansyl derivative-based (DIlSD) fluorescence probe was designed and synthesized. Its combination with anionic surfactant SDS assemblies shows enhanced fluorescence intensity and blue-shifted maximum wavelength. Its fluorescence can be slightly quenched by Cu(2+); however, the fluorescence quenching efficiency by Cu(2+) is highly increased upon titration of arginine (Arg). As a result, the ternary system containing the cationic fluorophore, anionic surfactant, and Cu(2+) functions as a highly sensitive and selective sensor to Arg. The optimized sensor system displays a detection limit of 170 nM, representing the highest sensitivity to Arg in total aqueous solution by a fluorescent sensor. Control experiments reveal that the imidazolium groups in the fluorophore, the anionic surfactant, and Cu(2+) all play important roles in the process of sensing Arg. The electrostatic interaction between the cationic fluorophore and anionic surfactants facilitates the binding of imidazolium rings with Cu(2+), the surfactant surface-anchored Cu(2+) is responsible for further binding of Arg, and the electrostatic interaction between anionic surfactants and positively charged amino acids accounts for the selective responses to Arg.

Topics: Arginine; Chemistry Techniques, Analytical; Copper; Dansyl Compounds; Fluorescent Dyes; Imidazoles; Limit of Detection; Sodium Dodecyl Sulfate; Solutions; Surface-Active Agents; Water

An automated methodology for the kinetic study of β-galactosidase activity in sodium dodecylsulfate (SDS)/ionic liquid (IL) mixed micelles was developed. The main objective of the work was the evaluation of mixed micelles as reaction media for the industrial synthesis of glyco-oligossacharides. Enzyme activity was evaluated by means of a model reaction with the fluorescent substrate 4-methylumbelliferyl-α-d-galactopyranoside (MUG). The assay was implemented in a sequential injection analysis (SIA) system and enzyme activity was studied in SDS/bmim [BF(4)] and SDS/hmim [Cl] mixed micelles with variable concentrations of both components. In order to perform a critical evaluation of the obtained results, CMC and average micellar size of SDS/hmim [Cl] mixed micelles were evaluated by fluorescence and dynamic light scattering, respectively. In the micelle characterization assays it was observed that the CMC of the mixed micelles increased with hmim [Cl] concentration up to 1molL(-1). In the presence of higher IL concentrations there were no evidences of micelle formation. Regarding micellar size, it was maximum for an IL concentration of 0.09molL(-1). The kinetic assays evidenced that SDS/bmim [BF(4)] and SDS/hmim [Cl] mixed micellar systems can led to an increase of enzyme activity. This increase is dependent on the variation of the average micellar size that occurs with the increase of IL concentration up to 0.09molL(-1). It was also noticed that the most promising systems are those incorporating SDS and IL in concentrations under 50mmolL(-1) and 0.5molL(-1), respectively(.) These results evidenced that the studied ILs can modify the physico-chemical properties of the surfactant solution in a favourable way regarding β-galactosidase activity being an important achievement for the future implementation of industrial processes catalyzed by this enzyme, mainly the synthesis of glyco-oligossacharides. Indeed, surfactant/IL mixed micelles proved to be an interesting alternative to conventional organic solvents in this field enabling the implementation of the processes in a relatively hydrophobic media with enhanced enzyme activity.

Topics: Aspergillus oryzae; beta-Galactosidase; Biotechnology; Enzyme Assays; Flow Injection Analysis; Imidazoles; Ionic Liquids; Kinetics; Micelles; Sodium Dodecyl Sulfate

A biomimetic was designed for the construction of a new efficient peroxidase-like nano artificial enzyme with a heme-imidazole component complexed with gemini 12-2-12/SDS supramolecules. The presence of a simple surfactant mixture (SDS/gemini 12-2-12 at a particular concentration) provided an apoprotein-like hydrophobic pocket for the heme-imidazole moiety, which produced a peroxidase active site containing positive and negative charges distributed on the colloidal surface. Vesicular structures that stabilized the heme-imidazole complexes formed multienzyme advanced colloids. The enzymatic activation parameters indicated that the catalytic efficiency of the novel nano artificial enzyme was 27% as efficient as the native horseradish peroxidase (HRP). The imidazole moiety, which functionally corresponded to the histidine ligand in the native HRP, increased the reactivity and catalytic efficiency of the artificial enzyme. The nano biocatalyst did not exhibit suicide inactivation until high concentrations of hydrogen peroxide, indicating that the vesicle hydrophobic pocket effectively shielded the active site, thereby controlling the concentration of hydrogen peroxide at the heme moiety and enabling high rates of enzymatic turnover.

Topics: Apoproteins; Biocatalysis; Biomimetic Materials; Calcitriol; Catalytic Domain; Hemin; Horseradish Peroxidase; Hydrophobic and Hydrophilic Interactions; Imidazoles; Kinetics; Micelles; Nanotechnology; Sodium Dodecyl Sulfate; Spectrophotometry, Ultraviolet

The active site of several oxygen binding proteins can be mimicked with the ferric iron protoporphyrin IX derivative hemin, coordinating two imidazole molecules and embedded in sodium dodecyl sulfate (SDS) micelles; the detergent simulates the hydrophobic cavity of heme proteins. We studied the low-frequency vibrational modes of the porphyrin-iron-imidazole bonding in infrared absorbance spectra. Assignment of the metal-ligand vibrations to signals at 396, 387, and 378 cm(-1) was performed by isotope labeling of the imidazole ligand. These modes were also found to be temperature-dependent and to display a linear increase of signal intensity between 25 and 150 K and, with a different slope, between 150 and 300 K. The modes at 396 and 399 cm(-1) show for 25 K an up-shift about 4 cm(-1) and the signal at 378 cm(-1) a small downshift, indicating the involvement of antisymmetric stretching modes and, in the latter, of bending motions. Anharmonic couplings to doming modes are discussed, and the doming mode and hydrogen-bonding signature spectral range between 300 and 100 cm(-1) is presented.

Topics: Biomimetic Materials; Heme; Hydrogen Bonding; Imidazoles; Iron; Isotope Labeling; Ligands; Metalloproteins; Micelles; Protoporphyrins; Sodium Dodecyl Sulfate; Spectrophotometry, Infrared; Surface-Active Agents; Temperature

A simple micellar electrokinetic chromatography (MEKC) method with UV detection at 200 nm for analysis of piracetam in plasma and in cerebrospinal fluid (CSF) by direct injection without any sample pretreatment is described. The separation of piracetam from biological matrix was performed at 25 degrees C using a background electrolyte consisting of Tris buffer with sodium dodecyl sulfate (SDS) as the electrolyte solution. Several parameters affecting the separation of the drug from biological matrix were studied, including the pH and concentrations of the Tris buffer and SDS. Under optimal MEKC condition, good separation with high efficiency and short analyses time is achieved. Using imidazole as an internal standard (IS), the linear ranges of the method for the determination of piracetam in plasma and in CSF were all between 5 and 500 microg/mL; the detection limit of the drug in plasma and in CSF (signal-to-noise ratio=3; injection 0.5 psi, 5s) was 1.0 microg/mL. The applicability of the proposed method for determination of piracetam in plasma and CSF collected after intravenous administration of 3g piracetam every 6h and oral administration 1.2g every 6h in encephalopathy patients with aphasia was demonstrated.

Topics: Adult; Chromatography, Micellar Electrokinetic Capillary; Humans; Hydrogen-Ion Concentration; Imidazoles; Piracetam; Reproducibility of Results; Sodium Dodecyl Sulfate; Tromethamine

Topics: Acrylic Resins; Electrophoresis, Polyacrylamide Gel; Imidazoles; Sensitivity and Specificity; Sodium Dodecyl Sulfate; Staining and Labeling; Zinc Sulfate

Film-entrapped myoglobin exhibits well-defined electrochemistry which, upon ligand binding, displays a titratable redox potential shift. This effect has been observed to be highly dependent on the charged state of involved films. We have demonstrated that this approach may act as a model system for studies of molecular recognition between proteins and ligands.

Topics: Cetrimonium; Cetrimonium Compounds; DNA; Electrochemistry; Imidazoles; Ligands; Macromolecular Substances; Membranes, Artificial; Myoglobin; Oxidation-Reduction; Phosphatidylcholines; Polyethyleneimine; Protein Binding; Sodium Dodecyl Sulfate

The sensitivity, simplicity, and relative rapidity of Coomassie blue staining have made this technique the method of choice for routine detection and quantitative analysis of gel electrophoresis-separated protein bands in many applications. To extend the usefulness of this technique, we have developed a new double-staining method for visualizing SDS-PAGE-separated protein bands that were undetected by Coomassie blue staining of the gel. Coomassie blue-stained gels are washed in distilled water (15 min, two times) and then subjected to imidazole-zinc reverse staining. As a result of the method, a homogeneous white-stained background is generated and two types of protein bands can be observed: (a) typical Coomassie blue-stained bands, which appear superposed on larger transparent bands; and (b) reverse-stained (transparent) bands, which were previously undetected by the Coomassie blue staining. The method is rapid, simple, and reproducible and double-staining gels can be kept in distilled water for months without loss of the protein pattern. The overall sensitivity is high (e.g., 1.6 ng for recombinant streptokinase, 47 kDa) over a wide range of protein molecular weights (10 to 100 kDa) and independent of the degree of Coomassie blue destaining of the gel. Furthermore, a mechanism offering a consistent explanation for the role of imidazole, SDS, and zinc in the reverse staining of gels, particularly after Coomassie blue staining is proposed.

Topics: Electrophoresis, Polyacrylamide Gel; Imidazoles; Proteins; Rosaniline Dyes; Sodium Dodecyl Sulfate; Staining and Labeling; Zinc

A reverse staining procedure is described for the detection of proteins in acrylamide and agarose gels with and without SDS. Protein detection occurs a few minutes after electrophoresis. The sensitivity on acrylamide gels is higher than that of Coomassie blue staining either on acrylamide gels or on electrotransferred membranes. Sequencing of protein bands only detected by reverse staining on the gel and not by Coomassie blue is demonstrated.

Topics: Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Imidazoles; Proteins; Sodium Dodecyl Sulfate; Staining and Labeling; Zinc