Polyphenol oxidase (PPO) -catalyzed browning reactions are of significant importance in the fruit and vegetable industry. These reactions proceeding in many foods of plant origin cause deterioration and loss of food quality. A better knowledge of the factors that influence the action of PPO is imperative in order to control and manipulate its detrimental activity in plant products. This paper presents an overview of the current understanding of the reaction properties, biochemical characteristics and potential physiological roles of PPO in plants. Reaction properties will include general PPO reactions, specificities and molecular mechanisms of these reactions, and methods available to assess PPO activity. Physicochemical properties will evaluate substrate specificity, environmental influences such as pH and temperature, multiplicity, latency, and activators and inhibitors of PPO. The discussion will conclude with potential physiological roles of PPO in plants. 362 R. YORUK and M.R. MARSHALL PROPERTIES AND ROLE OF PPO 363will evaluate substrate specificity, environmental influences such as pH and temperature, multiplicity, latency, and activators and inhibitors of PPO. IMPORTANCE OF PPO IN THE FOOD INDUSTRY ProblemThe catalytic action of PPO has an enormous impact on the quality of several fruit and vegetable crops and results in alteration of color, flavor, texture, and nutritional value (Vamos-Vigyazo 1981). It is a limiting factor in the handling and technological processing of crops as peeled, sliced, bruised or diseased tissues rapidly undergo browning. Some commercially important edible plant products susceptible to adverse browning reactions include fruits such as apple (Harel er al.

The potential mode of inhibition for oxalic acid on polyphenol oxidase (PPO) was investigated. The extent of inhibition was influenced not only by oxalic acid concentration but also by pH. Inhibition was most prominent at pH 4.0 where complete inhibition occurred at the 4-mM oxalic acid concentration and was less evident at higher pH values. Inhibition of PPO by oxalic acid was due to its binding with copper to form an inactive complex, and the inhibition was characterized as noncompetitive. Oxalic acid diminished the catecholquinone product formation, and no quinone bleaching was observed. Oxalic acid was a more potent inhibitor of PPO compared with other structurally related acids.

Relative antibrowning potency of oxalic acid on banana and apple slices was investigated using a machine vision system. Degree of browning on fresh-cut surfaces was evaluated visually and quantitatively by observing changes of CIE L* values and evaluating temporal changes in color spectra based on experimental variables, oxalic acid concentration, and storage time. Browning inhibition was most prominent on banana and apple slices treated with oxalic acid solutions at concentrations of 60 and 5 mM, respectively. Oxalic acid was a more potent antibrowning agent compared with other structurally related acids. Average residual oxalic acid levels in the tissues for an effective antibrowning activity were measured.

A new natural apple polyphenol oxidase (PPO) inhibitor(s) from housefly (Musca domestica L.) was discovered. Crude inhibitor(s) isolated by buffer extraction, heat treatment, and dialysis from housefly pupae inhibited the activity of apple PPO up to 90% at pH values above 5.0. Inhibition was strictly pH-dependent. The inhibitor(s) was further characterized by employing heat, freezing and thawing, irradiation, pH adjustment, and ultrafiltration studies. The potential PPO inhibitor(s) was stable to heating at 100 °C for 1 h, repeated freezing and thawing, and irradiation. The inhibitor(s) was most stable at pH around 5.0 and least stable at alkaline pH. The PPO inhibition profile of housefly during metamorphosis also was evaluated.

The importance of maintaining stable pH when studying antibrowning activity of oxalic acid in a catechol‐polyphenol oxidase model system was demonstrated. Enzyme inhibition in the reaction medium showing a decrease in pH caused by the addition of increasing oxalic acid concentrations was compared with the inhibition of the same oxalic acid concentrations at a stable pH of 5.0. Relative antibrowning activity of oxalic acid was significantly lower at stable pH conditions than at variable pH conditions. The main reason for this was because of the very slight alterations around the pH used for the experiments, which were sufficient to induce significant differences in the catalytic activity of the enzyme. Oxalic acid at pH 5.0 was found to be a noncompetitive inhibitor of the mixed‐type with an inhibitor constant (Ki) of 0.8 mM. PRACTICAL APPLICATIONS Fruits and vegetables are prone to polyphenol oxidase (PPO)‐catalyzed browning reactions during handling and processing, and the food industry is interested in methods to prevent browning for minimally processed crops. Although not an approved food additive, oxalic acid is a common component of several foods from plant origin. There are few reports on the inhibition of PPO by oxalic acid in reaction model systems. The importance of maintaining a stable pH when studying the antibrowning activity of oxalic acid in a catechol‐PPO model system was demonstrated. Small additions of oxalic acid into the assay system induced significant pH variation. Minute changes in the degree of acidity severely affected PPO activity and resulted in significant errors in measuring inhibition. Identifying how environmental changes in assay conditions affect activity and inhibition are important in interpreting how this enzyme is controlled on foods by inhibitors.