Effect of temperature and pressure on yeast invertase stability: a kinetic and conformational study

Abstract

Kinetics of the temperature- or pressure-induced denaturation of invertase from Saccharomyces cerevisiae were obtained in the temperature range 45-70 degrees C and in the pressure range 500-650 MPa. The investigation was done by measuring the residual activities after cooling or pressure release and the intrinsic fluorescence of aromatic amino-acids (tyrosine and tryptophan) upon excitation at 277 nm. The residual activity decreased exponentially as a function of time incubation according to a biphasic model either with pressure or temperature, whereas the fluorescence emission indicated a difference between these two parameters. When the enzyme was subjected to thermal treatment, the fluorescence of tyrosine and tryptophan decreased slowly, while after high-pressure treatment, these aromatic residues become more exposed to the aqueous solvent during unfolding, giving rise to a large decrease in fluorescence in the 330-340 nm region. Moreover, in the latter case, an enhancement of light scattering intensity showed changes in protein-protein interactions.