Active Site Crowding of Cytochrome P450 3A4 as a Strategy To Alter Its Selectivity

Substrate‐promiscuous enzymes are a promising starting point for the development of versatile biocatalysts. In this study, human cytochrome P450 3A4, known for its ability to metabolise hundreds of drugs, was engineered to alter its regio‐ and stereoselectivity. Rational mutagenesis was used to introduce steric hindrance in a specific manner in the large active site of P450 3A4 and to favour oxidation at a more sterically accessible position on the substrate. Hydroxylation of a synthetic precursor of (R)‐lisofylline, a compound under investigation for its anti‐inflammatory properties, was chosen as a first proof‐of‐principle application of our protein engineering strategy. In a second example, increasing active site crowding led to an incremental shift in the selectivity of oxidation from an internal double bond to a terminal phenyl group in a derivative of theobromine. The same correlation between crowding and selectivity was found in a final case focused on the hydroxylation of the steroid sex hormone progesterone.     

Collective dynamics modeling of polydisperse particulate systems via Markov chains

This paper develops an efficient approach to modeling and analyzing the overall dynamics of polydisperse particulate systems, exemplified using a rotating drum with horizontal axis, under both constant and time-varying operating conditions. This approach captures the collective dynamics using stochastic models in the form of Markov chains. The characteristics of such dynamics can be obtained from the Markov chains operator. It provides a systematic way to the analysis of collective dynamical features of particle movements. The obtained operators are used to estimate the spatial particle distribution and the degree of particulate mixing as examples of collective dynamic features of polydisperse particulate systems. In this paper, Markov chains models were developed from discrete element method simulation results to show the effectiveness of the proposed approach.     

Sensitiveness of Porous Silicon‐Based Nano‐Energetic Films

Nanoporous silicon (pSi) films on a silicon wafer were loaded with sodium perchlorate and perfluoropolyether (PFPE) oxidizing agents. Sensitiveness to impact, friction and electrostatic discharge (ESD) of the resulting energetic thin films were investigated. It was observed that pSi loaded with perchlorate was sensitive at the lowest limit of detection for the available equipment (<4.9 J impact energy, <5 N friction force, and <45 mJ ESD spark energy). When loaded with PFPE the material was very sensitive to impact (<4.9 J), moderately sensitive to ESD (between 45 and 100 mJ) and insensitive to friction (>360 N). pSi loaded with either perchlorate or PFPE displayed behavior during sensitiveness testing similar to other primary explosive materials.     

Dual‐Layer Hollow Fibers for Sulfur Removal from Fuels

Emission of sulfur compounds to the atmosphere is universally recognized as one key target to be reduced. For membrane pervaporation which is considered as a potential purification process of fuels, dual‐layer polyurethane (PU)/polyethersulfone hollow‐fiber membranes were prepared. A novel fabrication technique is proposed using a quadruple spinneret to produce the fiber with such morphology by simultaneous spinning of two polymer solutions in the presence of two corresponding precipitation media. Activated carbon was added into the PU solution to improve the transport properties of the selective layer. Resulting hollow‐fiber membranes showed very good adhesion between the selective layer and its support, in addition to an effective removal of a sulfur compound such as 2‐methyl thiophene from a typical model fuel, an indication of good prospects for both the fabrication technique and for sulfur removal by pervaporation of fuels.     

Cuticular Hydrocarbons as Sex Pheromone of the Bee <Emphasis Type="Italic">Colletes cunicularius</Emphasis> and the Key to its Mimicry by the Sexually Deceptive Orchid, <Emphasis Type="Italic">Ophrys exaltata</Emphasis>

Male Colletes cunicularius bees pollinate the orchid, Ophrys exaltata, after being sexually deceived by the orchid’s odor-mimicry of the female bee’s sex pheromone. We detected biologically active volatiles of C. cunicularius by using gas chromatographic–electroantennographic detection (GC-EAD) with simultaneous flame ionization detection. After identification of the target compounds by coupled gas chromatography–mass spectrometry (GC-MS), we performed behavioral tests using synthetic blends of the active components. We detected 22 EAD active compounds in cuticular extracts of C. cunicularius females. Blends of straight chain, odd-numbered alkanes and (Z)-7-alkenes with 21–29 carbon atoms constituted the major biologically active compounds. Alkenes were the key compounds releasing mating behavior, especially those with (Z)-7 unsaturation. Comparison of patterns of bee volatiles with those of O. exaltata subsp. archipelagi revealed that all EAD-active compounds were also found in extracts of orchid labella. Previous studies of the mating behavior in C. cunicularius showed linalool to be an important attractant for patrolling males. We confirmed this with synthetic linalool but found that it rarely elicited copulatory behavior, in accordance with previous studies. A blend of active cuticular compounds with linalool elicited both attraction and copulation behavior in patrolling males. Thus, linalool appears to function as a long-range attractant, whereas cuticular hydrocarbons are necessary for inducing short-range mating behavior.     

Analytical investigation of the composition of plasma-induced functional groups on carbon nanotube sheets

To increase the applicability of carbon nanotubes (CNTs) oxygen-containing functional groups were generated on their widely inert surface by using glow-discharge plasmas. CNT-sheets (bucky papers) produced from the powder-like raw material were used as substrates allowing for a more defined characterization of one and the same surface by different analytical techniques. The plasma composition was analyzed by optical emission spectroscopy. Since the actual composition of the plasma-induced functional groups has still not been completely resolved, we performed an in-depth characterization of the treated samples by X-ray photoelectron (XPS) and Raman spectroscopy as well as electron spin resonance measurements. To overcome limitations of the XPS-analysis in distinguishing between groups featuring nearby binding energies, alcohol-, keto-/aldehyde-, and carboxyl-groups were tagged by derivatization techniques using fluorine-containing reagents (trifluoroaceticanhydride, trifluoromethylphenylhydrazine, and trifluoroethanol). Differential spectra were calculated to enhance the accuracy of the deconvolution of the XPS-spectra. This enabled us to determine dependencies of the plasma parameters, i.e. treatment time, process pressure, and gas composition (mixtures of Ar, O₂, H₂O, and H₂), on the composition of the generated functional groups as well as an up to 6-fold enhancement in derivatizable groups for switching process gas from Ar/O₂ to Ar/H₂O.     

Method for Accurate Surface Temperature Measurements During Fast Induction Heating

A robust method is proposed for the measurement of surface temperature fields during induction heating. It is based on the original coupling of temperature-indicating lacquers and a high-speed camera system. Image analysis tools have been implemented to automatically extract the temporal evolution of isotherms. This method was applied to the fast induction treatment of a 4340 steel spur gear, allowing the full history of surface isotherms to be accurately documented for a sequential heating, i.e., a medium frequency preheating followed by a high frequency final heating. Three isotherms, i.e., 704, 816, and 927°C, were acquired every 0.3 ms with a spatial resolution of 0.04 mm per pixel. The information provided by the method is described and discussed. Finally, the transformation temperature Ac1 is linked to the temperature on specific locations of the gear tooth.