Full Thermal Switching of Enzymes by Thermoresponsive Poly(2-oxazoline)-Based Enzyme Inhibitors

Controlling the activity of enzymes is an important feature for many processes in medicine, bioanalytics, and biotechnology. So far, it has not been possible to fully switch biocatalysts on and off by thermoresponsive enzyme inhibitors. Herein, we present poly(2-oxazoline)s with iminodiacetic acid end groups (POx-IDA) that are lower critical solution temperature (LCST) polymers and thus thermosensitive. They are capable of reversibly inhibiting the activity of horse radish peroxidase and laccase by more than 99 %. Increasing the temperature makes the POx-IDA precipitate, which leads to 100 % recovery of the enzyme activity. This switching cycle is fully reversible. The LCST of the POx-IDA can be tuned by varying the polymer composition to generate a wide range of switching windows.     

Reconstruction of the (111)-surfaces of homopolar semiconductors: A nonlinear polarizability model

The phases of the (111)-surface of C, Si and Ge are studied. In the top two layers harmonic forces and bond polarizabilities are considered. The calculated 7×7 reconstruction of Si shows good agreement with experiments. The resulting 2×1 structure is compared with that predicted by other treatments. Our model gives a transition from reconstructed to 1×1 disordered phases at finite temperatures as observed in Si and Ge.     

Wall Recombination in Glow Discharges

We derived a model describing the radial distribution of the ion and electron densities and the radial electric field strength in the positive column of a glow discharge. The set of equations related to the plasma consists of the equations for particle and momentum conservation for the ions and electrons and the Poisson-equation. In a novel approach, the necessary boundary conditions in our model result from a system of balance equations for the charge carriers on the insulated wall surrounding the positive column.     

Surface-induced hydrogelation

The first azo dye that gels from its aqueous solution was synthesized and its moist hydrogel was investigated with electron microscopy and atomic force microscopy; the anionic azo dye in aqueous solution forms a hydrogel on cationic surfaces even at concentrations 50 times below the minimal gelation concentration.     

Measurements of the solubilities of derivatized amino acids in supercritical carbon dioxide

The solubilities of phenylalanine and tyrosine in supercritical carbon dioxide (SCCO₂) were measured after derivatization as the N-acetyl amino acid ethyl ester, N-carbobenzoxy amino acid and N-acetyl amino acid. Using an SCCO₂ flow system, a measuring method of the saturated solubilities of the derivatized amino acids was established in which the contact height of the extraction cell, i.e. a packed column, is increased till the concentration of a derivatized amino acid at the exit of the cell reaches a plateau. The solubilities of N-acetyl phenylalanine ethyl ester (APEE) exceeded 0.001 mole fraction, which is higher than those of caffeine produced in industrial SCCO₂ processes. A possible way of separating the amino acid mixtures using polarity differences in different amino acid side chains was demonstrated using the solubility data of the N-acetyl-amino acid ethyl esters in SCCO₂, as the solubilities of APEE are higher than those of N-acetyl tyrosine ethyl ester by two orders of magnitude.     

Theoretical and experimental studies of biotin analogues that bind almost as tightly to streptavidin as biotin

We have used a newly developed qualitative computational approach, PROFEC (Pictorial Representation of Free Energy Changes), to visualize the areas of the ligand biotin where modifications of its structure might lead to tighter binding to the protein streptavidin. The PROFEC analysis, which includes protein flexibility and ligand solvation/desolvation, led to the suggestion that the pro-9R hydrogen atom of biotin, which is in alpha-position to the CO(2)(-) group, might be changed to a larger group and lead to better binding with streptavidin and avidin. Free energy calculations supported this suggestion and predicted that the methyl analogue should bind approximately 3 kcal/mol more tightly to streptavidin, with this difference coming exclusively from the relative desolvation free energy of the ligand. The PROFEC analysis further suggested little or no improvement for changing the pro-9S hydrogen atom to a methyl group, and great reduction in changing the ureido N-H groups to N-CH(3). Stimulated by these results, we synthesized 9R-methylbiotin and 9S-methylbiotin, and their binding free energies and enthalpies were measured for interaction with streptavidin and avidin, respectively. In contrast to the calculated results, experiments found both 9-methylbiotin isomers to bind more weakly to streptavidin than biotin. The calculated preference for the binding of the 9R- over the 9S-stereoisomer was observed. In addition, 9-methylbiotin is considerably less soluble in water than biotin, as predicted by the calculation, and the 9R isomer is, to our knowledge, thus far the tightest binding analogue of biotin to streptavidin. Subsequently, X-ray structures of the complexes between streptavidin and both 9R- and 9S-methylbiotin were determined, and the structures were consistent with those used in the free energy calculations. Thus, the reason for the discrepancy between the calculated and experimental binding free energy does not lie in unusual binding modes for the 9-methylbiotins.     

Synthese der citromycinone stereoselektive öffnung eines epoxids zum cis-diol

The citromycinone-derivatives $\text{14}$ and $\text{15}$ have been obtained by intramolecular Marschalk cyclisation of α-hydroxyaldehyde $\text{12}$. An unexpected opening of epoxide $\text{16}$ to $\text{cis}$-diol $\text{14}$ was observed. By exclusion of the alternative formula $\text{2}$ structure $\text{3}$ is proposed for α-citromycinone.     

A solid phase linker strategy for the direct synthesis of EDANS-labelled peptide substrates

A novel linker strategy for the efficient synthesis of peptides C-terminally labelled with the EDANS fluorophore is described. Using this support, FRET peptide substrates bearing EDANS/Dabcyl fluorescent donor/acceptor groups can be readily prepared using standard Fmoc solid phase methods.     

Experimental design-based development and single laboratory validation of a capillary zone electrophoresis method for the determination of the artificial sweetener sucralose in food matrices

A capillary zone electrophoresis (CZE) method, optimised chemometrically, underwent a complete in-house validation protocol for the qualification and quantification of sucralose in various foodstuffs. Separation from matrix components was obtained in a dinitrobenzoic acid (3 mM)/sodium hydroxide (20 mM) background electrolyte with a pH of 12.1, a potential of 0.11 kV cm^–1 and a temperature of 22 °C. Detection was achieved at 238 nm by indirect UV. Screening, optimisation and robustness testing were all carried out with the aid of experimental design. Using standard addition calibration, the CZE method has been applied to still, carbonated and alcoholic beverages, yoghurts and hard-boiled candy. The method allows the detection of sucralose at >30 mg kg^–1, with a linearity range of 50–500 mg kg^–1, making it suitable for implementation of the recently amended Sweeteners for use in foodstuffs Directive (European Parliament and Council (2003) Off J L237:3–12), which set maximum usable doses of sucralose for many foodstuffs, most ranging from 200 mg kg^–1 to 450 mg kg^–1.