No abstract

The influence of polyhydric alcohols and carbohydrates on the thermostability, i.e., the heat inactivation kinetics, of Bacillus licheniformis alpha-amylase was studied in the temperature range 96 degrees to 130 degrees C. High concentrations (from 9 to 60 weight percent) of glycerol, sorbitol, mannitol, sucrose, or starch can markedly decrease the inactivation rate constant, k, and in the studied cases, this stabilizing effect grows stronger with increasing additive concentration. Statements about stabilization should, however, be specified carefully with respect to temperature, because E(A) is mostly altered likewise. For dissolved enzyme E(A) was almost always decreased in the presence of polyol or carbohydrate, whereas for immobilized enzyme it was augmented in each studied instance. The inactivation of dissolved enzyme can, in all the studied cases, be adequately described as a first-order process. Immobilized enzyme, however, shows biphasic then first-order inactivation kinetics, depending on the additive concentration and temperature.

Measurements of residual enzyme activity and FTIR-spectroscopy revealed that mushroom polyphenoloxidase is a thermosensitive enzyme readily inactivated by temperatures exceeding 50ЊC. The enzyme is, however, very pressure stable. At room temperature no enzyme inactivation occurred at pressures up to 6 kbar. FTIR-spectroscopy revealed that a modification in enzyme conformation occurred below 9 kbar. Kinetic studies showed that pressure and temperature did not always act synergistically with respect to enzyme inactivation. FTIR-spectroscopy showed that pressure-or temperature-induced changes were considerably different. Both temperature and pressure-temperature inactivation were only slightly influenced by pH (in the pH-range 5.5-7.5).

In this review, current methods used to evaluate the integrated impact of time and temperature upon preserving a food product by a heat treatment are considered. After identifying the basic premise any preservation scheme shall meet, the central role of a feasible description for the heat activation kinetics of microorganisms, their spores, and other quality attributes are stressed. Common concepts to quantify a thermal process are presented. Shortcomings of the prevalent evaluation methods are highlighted and attention is given to the development, restrictions, and possibilities of time-temperature-integrators as "new" evaluation tools to measure the impact of a "classical" in-pack heat treatment and more modern heating techniques such as continuous processing of solid/liquid mixtures on foods.

In view of a possible application of the alpha-amylase from Bacillus licheniformis as a time-temperature integrator for evaluation of heat processes,(11) thermal inactivation kinetics of the dissolved and covalently immobilized enzyme were studied in the temperature range 90-108 degrees C. The D-values (95 degrees C) for inactivation of alpha-amylase, dissolved in tris-HCl buffer, ranged from 6 to 157 min, depending on pH, ionic strength, and Ca(2+) and enzyme concentration. The z-value fluctuated between 6.2 and 7.6 degrees C. On immobilization of the alpha-amylase by covalent coupling with glutaraldehyde to porous glass beads, the thermoinactivation kinetics became biphasic under certain circumstances. For immobilized enzyme, the D-values (95 degrees C) ranged between 17 and 620 min, depending largely on certain environmental conditions. The z-value fluctuated between 8.1 and 12.9 degrees C. In each case of biphasic inactivation, the z-value of the stable fraction (with the higher D-values) was lower than the z-value of the labile fraction.