Polyphenol oxidases in plants

Polyphenol oxidases and peroxidases are among the most studied enzymes in fruits and vegetables. Owing to the deleterious effects of discoloration and off-flavor formation induced by their actions, these enzymes have not ceased to be a matter of concern to food technologists, while their versatility as catalyst and their diversity as protein present a challenge to the biochemist. This article gives an account on the present state of knowledge in this field. The occurrence of polyphenol oxidases and peroxidases in food and food raw materials, and their role and importance in food processing are briefly outlined. Results of biochemical research including catalytic properties, substrate specificity, susceptibility towards pH and temperature, action of inhibitors, isolation, purification, and characteristics of the enzymes are given, with special emphasis on recent achievements based on high resolution separation and isoenzyme techniques. Finally, the behavior of polyphenol oxidase and peroxidase in selected major groups of fruits and vegetables is discussed. Some contradictions found in the literature are pointed out and some questions that have not been given the necessary attention by researchers so far are mentioned.

Polyphenol oxidase and peroxidase in fruits and vegetables.

This review examines the parameters of enzymatic browning in apple and apple products that is, phenolic compounds, polyphenoloxidases, and other factors (ascorbic acid and peroxidases), both qualitatively and quantitatively. Then the relationships between intensity of browning and the browning parameters are discussed, including a paragraph on the methods used for browning evaluation. Finally, the different methods for the control of browning are presented.

Enzymatic browning reactions in apple and apple products

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.

Physicochemical properties and function of plant polyphenol oxidase: a review'

The polyphenol oxidase system in clingstone peach (Prunus persica) was investigated. Polyacrylamide disc-gel electrophoresis indicated four bands with polyphenol oxidase activity in extracts from acetone powder of clingstone peach. These four isozymes were then isolated from a buffer extract of peach acetone powder by cold acetone precipitation, followed by diethylaminoethyl cellulose column chromatography. All isozymes had different heat stabilities. At 55 C, polyphenol oxidases A, B, and D had half-lives of 5.4, 14.6, and 14.1 minutes, respectively. Polyphenol oxidase C was stable over a period of 50 minutes of incubation at 55 C, but had a half-life of 2.2 minutes at 76 C. None of the isozymes had monophenolase activity, and they varied in their specificity for several diphenols. The following values were found for polyphenol oxidases A, B, C, and D, respectively, with catechol as substrate: optimal pH: 6.8, 6.5, 7.2, and 7.0; Michaelis constant: 6.6, 4.2, 7.0, and 36 mm; Vmax/(E0): 4.95, 39.4, 2.16, and 80.0 (ΔA min−1 mg−1). Each isozyme showed a different amount of inhibition by NaHSO3, NaCl, NaCN, l-ascorbic acid, glutathione, ethylenediaminetetraacetate, and sodium diethyldithiocarbamate.

Isolation and characterization of polyphenol oxidase isozymes of clingstone peach.

SUMMARY— The phenoloxidase system in the tissues of mushrooms, potatoes, and apples was investigated using a polyacrylamide electrophoretic technique. The enzyme system was shown to exhibit the phenomenon of multiple forms. The multiple form pattern obtained was highly characteristic for each individual species and variety studied, and substrate specificity was evident. The mushroom phenoloxidase system (Agaricus campestris) was shown to consist of at least nine distinct dl-dopa-reactive multiple forms, and at least three forms reacting with I-tyrosine. Potatoes (var. Rural Russet) showed at least 11 bands of dl-dopa activity, while apples (var. Golden Delicious) had at least three multiple forms of dl-dopa activity. By introducing the “multiphase” gel electrophoretic technique a better resolution of the multiple forms was obtained. A group of closely related dl-dopa multiple forms in mushrooms had the unique ability to withstand the temperature of 70°C for one hour. Sulfite, ethylenediamine-tetraacetic acid (EDTA) and other treatments affected the multiple forms differently. Each multiple form behaved as an individual entity upon repeated elutions and electrophoreses.

S. M. Constantinides

Papers

Multiple Forms of Phenoloxidase