Kinetic behavior of Desulfovibrio gigas aldehyde oxidoreductase encapsulated in reverse micelles

Biochemical and Biophysical Research Communications 308 (2003) 73–78We report the kinetic behavior of the enzyme aldehyde oxidoreductase (AOR) from the sulfate reducing bacterium Desulfovibrio gigas (Dg) encapsulated in reverse micelles of sodium bis-(2-ethylhexyl) sulfosuccinate in isooctane using benzaldehyde, octaldehyde, and decylaldehyde as substrates. Dg AOR is a 200-kDa homodimeric protein that catalyzes the conversion of aldehydes to carboxylic acids. Ultrasedimentation analysis of Dg AOR-containing micelles showed the presence of 100-kDa molecular weight species, confirming that the Dg AOR subunits can be dissociated. UV–visible spectra of encapsulated Dg AOR are indistinguishable from the enzyme spectrum in solution, suggesting that both protein fold and metal cofactor are kept intact upon encapsulation. The catalytic constant (kcat) profile as a function of the micelle size W0 (W0 ¼ ½H2O /[AOT]) using benzaldehyde as substrate showed two bell-shaped activity peaks at W0 ¼ 20 and 26. Furthermore, enzymatic activity for octaldehyde and decylaldehyde was detected only in reverse micelles. Like for the benzaldehyde kinetics, two peaks with both similar kcat values and W0 positions were obtained. EPR studies using spin-labeled reverse micelles indicated that octaldehyde and benzaldehyde are intercalated in the micelle membrane. This suggests that, though Dg AOR is found in the cytoplasm of bacterial cells, the enzyme may catalyze the reaction of substrates incorporated into a cell membrane

Subunit separation in reversed micelle system reveals the existence of active centers both on light and heavy γ-glutamyltransferase subunits

Regulation of supra-macromolecular composition and catalytic activity of a heterodimeric enzyme, γ-glutamyltransferase, in the system of Aerosol OT (sodium bis(2-ethylhexyl) sulfosuccinate) reversed micelles in octane were studied. Variation of the surfactant hydration degree (parameter, determining dimensions of the polar inner cavity of the micelle) causes a reversible dissociation of the enzyme to light and heavy subunits. Both enzyme subunits possess catalytic activity. The light and heavy subunits of the enzyme were separated on a preparative scale in a reversed micelle system using ultracentrifugation. The active centers of γ-glutamyltransferase were studied using its irreversible inhibitor - AT-125 (L·(αS.5S)-α-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid). Separation of the γ-glutamyltransferase subunits results in the 'opening' of a new active center located at the heavy subunit. In the dimer form of the enzyme this center is masked and it is not accessible to both substrate and inhibitor molecules. © 1991

Heat Flux Measurement in Shock Heated Combustible Gases and Clarification of Ignition Delay Time

Correct understanding of the ignition and combustion processes in the combustion chambers are critical for modeling advanced schemes of engines of high-speed aircraft and promising spacecraft. Moreover, experimental data on the ignition delay time are a universal basis for the development and testing of combustion kinetic models. Moreover, the higher the temperature of the fuel mixture, the smaller this time value and the more important its correct determination. The use of a thermoelectric detector allows to measure ignition delay times and record heat fluxes with a high time resolution (to tenths of μs) during ignition in propane–air mixtures. Due to the faster response time, the use of it allows refining the ignition delay time of the combustible mixture, and the detector itself can serve as a useful device that allows a more detailed study of the ignition processes

Nanostructured titanium-based materials for medical implants: Modeling and development

Nanostructuring of titanium-based implantable devices can provide them with superior mechanical properties and enhanced biocompatibity. An overview of advanced fabrication technologies of nanostructured, high strength, biocompatible Ti and shape memory Ni–Ti alloy for medical implants is given. Computational methods of nanostructure properties simulation and various approaches to the computational, ‘‘virtual’’testing and numerical optimization ofthese materials are discussed. Applications of atomistic methods, continuum micromechanics and crystal plasticity as well as analytical models to the analysis ofthe reserves ofthe improvement of materials for medical implants are demonstrated. Examples of successful development of a nanomaterial-based medical implants are presented.status: publishe