Impacts of high pressure assisted freezing on the denaturation of polyphenol oxidase

The mechanism of enzyme protein denaturation induced by high pressure freezing is complicated and unclear as this process involves Pressure-Factors (pressure and time) and Freezing-Factors (temperature, phase transition, recrystallization, and ice crystal types). In this study, the thermodynamics and conformation changes of mushroom polyphenol oxidase (PPO) under high pressure freezing treatments (HPF, ^{100,150,200,300,400,500MPa}P_-20°C/30min) and high pressure processes (HPP) followed with normal pressure immersion freezing (HPP-IF, ^100-500MPaP_25°C/30min - ^0.1MPaP_-20°C/30min) are investigated as compared with that processed under high pressure processes (HPP, ^100-500MPaP_25°C/30min) and normal pressure immersion freezing process (IF, ^0.1MPaP_-20°C/30min). The results suggested that the treated PPO with the same enzyme activity may have various thermodynamic characteristics and conformations; Pressure-Factors play the main roles in the denaturation of the PPO during the HPF treatment, and Freezing-Factors can weak the effect of Pressure-Factors on PPO denaturation; The treated PPO may be transferred into a partially fold intermediate state.