Enzyme stabilization: state of the art

Enzyme stabilization is one of the most important fields in basic and applied enzymology. In basic enzymology, it is of particular relevance to understand enzyme stabilization principles first elucidating how and why the enzymes lose their biological activity and then deriving structure-stability relationships existing in enzymatic molecules. In applied enzymology, the most significant goal is to achieve useful compounds by biocatalysis. Enzymes are good catalysts in terms of high catalytic and specific activity with ability to function under mild conditions. However, they are not always ideal catalysts for practical applications because they are generally unstable and they inactivate rapidly through several mechanisms. In order to enhance enzyme stability, many strategies have been pursued in recent years. The present article is an attempt to provide detailed information about these strategies.     

New experimental technique for detecting the effect of low-frequency electric fields on enzyme structure

A new experimental approach has been developed to determine kinetic and thermodynamic parameters of the inactivation of an enzyme under labile conditions both with and without exposure to electrical currents as sources of perturbation. Studies were undertaken to investigate if low-frequency electric currents can accelerate the thermal inactivation of an enzyme through interactions with dipole moments in enzymatic molecules and through related mechanical stresses. The experiments were conducted with the enzyme acid phosphatase. The enzyme was exposed to a 50-Hz current at different densities (10 to 60 mA/cm2 rms) or to a sinusoidal or square-wave current at an average density of 3 mA/cm2 and frequencies from, respectively, 50 Hz to 20 kHz and 500 pulses per second (pps) to 50,000 pps. Positive-control experiments were performed in the presence of a stabilizer or a deactivator. The results indicate that the technique is sensitive to conformational changes that otherwise may be impossible to detect. However, exposure to electric currents under the experimental conditions described herein showed no effects of the currents.     

Spontaneous and mitomycin-C-induced micronuclei in human lymphocytes exposed to extremely low frequency pulsed magnetic fields

The cytokinesis block micronucleus method, a very sensitive cytogenetic assay, was used to ascertain the possible genotoxic effects of extremely low frequency pulsed magnetic fields in phytohemagglutinin-stimulated human lymphocytes cultures from 16 healthy donors. Four conditions were studied: i) lymphocytes not exposed to the field (control cultures); ii) lymphocytes exposed to the field; iii) lymphocytes treated with mitomycin-C and not exposed to the field; iv) lymphocytes treated with mitomycin-C and exposed to the field. Mitomycin-C-treated cultures were used as control for the micronucleus method, because it is known that mitomycin-C is a potent genotoxic agent, capable of inducing micronuclei. The frequency of micronuclei in field-exposed cultures was similar to the spontaneous frequency observed in control unexposed-cultures. Moreover, the exposure to pulsed magnetic fields did not affect the frequency of micronuclei induced by mitomycin-C, suggesting that, in the experimental conditions used, this kind of field neither affected the integrity of chromosomes nor interfered with the genotoxic activity of mitomycin-C.