Kinetic studies on the multi-enzyme solution produced via solid state fermentation of waste bread by Aspergillus awamori

Abstract

The aim of this study was kinetic analysis of the multi-enzyme solution produced from waste bread via solid state fermentation by Aspergillus awamori. It was found that at normal temperature for hydrolysis reactions, 60°C, the activation energies for denaturation of A. awamori glucoamylase, 176.2kJ/mol, and protease, 149.9kJ/mol, are much higher than those for catalysis of bread starch, 46.3kJ/mol, and protein, 36.8kJ/mol. Kinetic studies showed that glucoamylase and protease in the multi-enzyme solution should have at least two conformations under the two temperature ranges: 30–55°C and 60–70°C. Thermodynamic analysis showed that, deactivation of glucoamylase and protease in the multi-enzyme solution can be reversible between 30°C and 55°C, since ΔS is negative and ΔH is positive. On the other hand, for glucoamylase and protease, both ΔS and ΔH are positive between 60°C and 70°C. This means that the deactivation of both enzymes in the multi-enzyme solution is spontaneous in this temperature range. It was also found that the glucoamylase produced in the solid state fermentation of waste bread is more thermally stable than the protease in the mixture. Consequently, the protease had little or no effect on the stability of the glucoamylase. Furthermore, the half-life of the glucoamylase produced from waste bread pieces was much higher than that produced from wheat flour. This is an important finding because the mode of production, via solid state fermentation, appears to have increased the thermostability of the enzyme significantly.