Purification, characterization and inactivation kinetics of polyphenol oxidase extracted from Cistanche deserticola

Abstract

PPO was purified from Cistanche deserticola, and its enzymatic characteristics were clarified. It was found that microwave treatment was an efficient way to inactivate PPO. Polyphenol oxidase (PPO) from Cistanche deserticola was obtained and purified through an acetone precipitation and anion exchange column, the enzymatic characteristics and inactivation kinetics of PPO were studied. The specific activity of PPO was 73135.15 ± 6625.7 U/mg after purification, the purification multiple was 48.91 ± 4.43 times, and the recovery was 30.96 ± 0.27%. The molecular weight of the PPO component is about 66 kDa by SDS-PAGE analysis. The optimum substrate of PPO was catechol (V_max = 0.048 U/mL, K_m = 21.70 mM) and the optimum temperature and pH were 30 °C and 7, respectively. When the temperature is above 50 °C, pH < 3 or pH > 10, the enzyme activity can be significantly inhibited. The first-order kinetic fitting shows that microwave inactivation has lesser k values, larger D values and shorter t_1/2. It was found that microwave treatment is considered as an efficient and feasible way to inactive PPO by comparing the Z values and E_a values of the two thermal treatments.

Keywords: Cistanche deserticola; Enzymatic properties; Inactivation of enzyme; Polyphenol oxidase; Purification.