Pulsed Light Inactivation of Mushroom Polyphenol Oxidase: a Fluorometric and Spectrophotometric Study

Polyphenol oxidase (PPO) is one of the most important food enzymes; it is responsible for the browning of many foods. Pulsed light (PL) is a non-thermal method of food preservation that is able to inactivate PPO. The aim of this work was to gain insight into the mechanism of PPO inactivation by PL. To this, the kinetics of PPO inactivation by PL was measured, together with associated changes in tryptophan fluorescence, KI fluorescence quenching, and turbidity; and results were analyzed by parameter A and phase diagram methods. Enzyme inactivation followed the Weibull model. Tryptophan fluorescence decreased during PL treatment, as well as the parameter A, while Stern-Volmer constants increased and turbidity was constant. The phase diagram showed only two populated states. There was a high correlation between the loss of activity and parameter A. Results indicate that under the experimental conditions, the inactivation of PPO by PL is an all-or-none process where the enzyme progressively unfolds with no evidence of aggregation.