CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering**

CYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16α-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternative binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that the entrance of water to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5α-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure–function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation.     

Mass transfer in trickle-bed reactors at low reynolds number

Mass transfer rates were determined in a 3.4 cm i.d. trickle-bed reactor in the absence of reaction by absorption measurements and in presence of reaction. Gas flow rates were varied from 0-100 l/h and liquid flow rates from 0-1.5 l/h. The catalyst particles were crushed to an average diameter of 0.054 and 0.09 cm. Mass transfer coefficients remained unaffected by change in gas flow rate but increased with liquid rate. The data from absorption measurements were evaluated with predictions based upon plug-flow and axial dispersion model. Mass transfer coefficients were found greater in case of axial dispersion model than that of plug-flow model specially at low Reynolds number (Re₁ < 1).Hydrogenation of α-methylstyrene to cumene using a Pd/Al₂O₃ catalyst was taken as a model reaction. Intrinsic kinetic studies were made in a laboratory-stirred-autoclave. Mass transfer coefficients were determined using these intrinsic kinetic data from the process kinetic measurements in trickle-bed reactor. Mass transfer coefficients under reaction conditions were found to be considerably higher than those obtained by absorption measurements.Correlations were suggested for predicting mass transfer coefficients at low Reynolds number.The gas to liquid mass transfer coefficients for lower gas and liquid flow rates were determined in a laboratory trickle-bed reactor. The effect of axial dispersion on mass transfer was considered in order to evaluate the experimental data. Three correlations were formulated to calculate the mass transfer coefficients, which included the effect of liquid loading, particle size and the properties of the reacting substances. The gas flow rate influences the gas to liquid mass transfer only in the region of low gas velocities. In the additional investigations of gas to liquid mass transfer without reaction in trickle-bed reactor, the mass transfer coefficients were determined under reaction conditions and the intrinsic kinetics was studied in a laboratory scale stirred autoclave with suspended catalyst. A few correlations are formulated for the mass transfer coefficients. A comparison with the gas-liquid mass transfer coefficient obtained by absorption measurements showed considerable deviations, which were illustrated phenomenologically.     

Induction plasma synthesis of fullerenes and nanotubes using carbon black–nickel particles

The existence of fullerenes (as allotropes of carbon) was established in the mid-1980s and during the last 15–18 years, systematic efforts have been devoted to improve the methods of their synthesis, including plasma-based system methods. The work presented here is focused on the investigation of fullerenes synthesis, using a radio frequency plasma reactor. The main objectives were to explore the use of induction plasma technology for the synthesis in-continuo of carbon fullerenes and to predict their formation conditions through conduct of theoretical studies. Thus, a thermodynamic study was carried out to predict the equilibrium composition of fullerenes produced at several combinations of operating conditions. Additionally, a statistical factorial design experiment, employing four factors at two levels, was also developed, in order to study the influence of the system’s operating parameters on the eventual C₆₀ fullerene yield. The results obtained showed that the reactor pressure, the electrical power and the raw material feed rates all have an important effect on the synthesis of fullerenes. The highest C₆₀ concentration in the products was found to be about 7.7 wt.%. Various other carbon nanostructures, such as nanotubes and nano-onions, were also successfully produced.     

Bestimmung des molekulargewichts von „mikrogelhaltigem”︁ polystyrol mit der ultrazentrifuge (methode archibald)

The weight average molecular weight M_w of a mixture of two polystyrenes with very high and relatively low molecular weights (M_w = 16.1 · 10⁶ and 98 · 10³ g/mole), as a model for polymers containing microgel, was determined by ultracentrifugation with the ARCHIBALD-methode. M_w decreases with increasing number of rotation N. The exact molecular weight can only be determined by extrapolation to N = 0. This value depends nearly completely on the molecular weight and weight percentage of the “microgel”. It is not suitable for the characterization of the whole sample.     

Ceramic Composites with Three-Dimensional Architectures Designed to Produce a Threshold Strength—II. Mechanical Observations

Finite element modeling and linear elastic fracture mechanics are used to model the residual stresses and failure stress of ceramic composites consisting of polyhedral alumina cores surrounded by thin alumina/mullite layers in residual compression. This type of composite architecture is expected to exhibit isotropic threshold strength behavior, in which the strength of the composite for a particular assumed flaw will be constant and independent of the orientation of tensile loading. The results of the modeling indicate that the strengths of such architectures will be higher than those of laminates of similar architectural dimensions that were previously found to exhibit threshold strength behavior for a particular flaw type. Flexural testing of the polyhedral architectures reveals that failure is dominated by processing defects found at junctions between the polyhedra. Fractography revealed the interaction of these defects with the residual stresses in the compressive layers that separate the polyhedra.     

Mechanical Behavior of Several Hybrid Ceramic-Matrix-Composite Laminates

Several different hybrid laminated composites comprised of alternating layers of dense ceramic sheets (either SiC or Si₃N₄) and fiber-reinforced ceramic-matrix-composite (CMC) layers (Nicalon fibers with either glass or glass-ceramic matrices) have been fabricated and characterized. The effects of the reinforcement architecture (unidirectional vs cross-ply) and the relative volume fractions of the phases on the tensile and flexural properties have been examined. Comparisons have been made with the properties of the constituent layers. Rudimentary models have been developed to describe the onset of cracking and for the minimum volume fraction of CMC required to develop multiple cracks and thus obtain a high failure strain.     

High-Strength Zirconia Fibers

Fine-grained polycrystalline zirconia fibers have been formed from an acetate precursor. The fibers contained a Y₂O₃ additive, which inhibited grain growth (grain size ≤0.5 μm) and allowed the tetragonal phase to be retained at room temperature. Fibers with diameters in the range 2 to 5 μm had strengths in the range 1.5 to 2.6 GPa.     

Effect of Zwitterionic Surfactants on Interparticle Forces, Rheology, and Particle Packing of Silicon Nitride Slurries

Phosphocholine (PC) zwitterionic surfactants, with different hydrocarbon chain lengths (C₆C₆PC to C₉C₉PC), were absorbed on the surface of silicon nitride near the isoelectric point (pH 6). Adsorption of the surfactants changed the lateral and normal surface forces, the rheology, and the consolidation behavior of the particles. The normal force between two silicon nitride surfaces as a function of separation and the lateral (friction) forces were measured using an atomic force microscope (AFM). These measurements indicated that surfactant adsorption reduced the magnitude of the long-range attractive van der Waals force and produced a repulsive short-range force. Although the adsorbed layers provided a barrier to particle contact, they could be ejected with a critical force that increased with the hydrocarbon chain length. The effect of an adsorbed layer on the viscosity and consolidation of slurries was also measured. The viscosity of all slurries decreased with increasing shear rate, indicative of attractive particle networks. The highest viscosity was observed for slurries formulated at the isoelectric point without added surfactant. Much lower viscosities were observed when the surfactant concentration was greater than the critical micelle concentration (cmc). A relative density of 0.46 was obtained via pressure filtration at 4 MPa without a surfactant, and between 0.46 to 0.59 (C₆C₆PC to C₉C₉PC, respectively) for surfactant concentrations greater than the cmc. Comparing force measurements with rheology and packing density provides a basis for discussing the role of interparticle forces in ceramic powder processing via colloidal routes.     

"Alumina" Surface Modification of Silicon Nitride for Colloidal Processing

Two different methods are used to coat silicon nitride particles with an alumina precursor to make Si₃N₄ behave like Al₂O₃ in aqueous slurries. The first method involves the precipitation of an aluminum hydroxycarbonate from dissolved Al(NO₃)₃ during the decomposition of urea. In the second method, dry silicon nitride powder is reacted with aluminum tri- sec -butoxide in hexane at room temperature. Both methods produce a coated powder in which the electrophoretic and rheological properties of aqueous slurries mimic those of alumina. When salt is added to slurries consisting of coated Si₃N₄ powder, all rheological evidence suggests the presence of a short-range repulsive potential that produces a weakly attractive particle network similar to that previously reported for Al₂O₃ powder. Although electrophoretic and rheological data showed that the coated powder behaved like Al₂O₃, consolidation data indicated that slurries of coated powder with added salt did not pack to high density. In addition, these bodies were not plastic as found for bodies consolidated from dispersed and salt-added Al₂O₃ slurries.