Purification and partial biochemical characterization of polyphenol oxidase from mango (Mangifera indica cv. Manila)

Polyphenol oxidase (PPO) is an enzyme widely distributed in the plant kingdom that has been detected in most fruits and vegetables. PPO was extracted and purified from Manila mango (Mangifera indica), and its biochemical properties were studied. PPO was purified 216-fold by hydrophobic interaction and ion exchange chromatography. PPO was purified to homogeneity, and the estimated PPO molecular weight (MW) by SDS-PAGE was ≈31.5 kDa. However, a MW of 65 kDa was determined by gel filtration, indicating a dimeric structure for the native PPO. The isolated PPO showed the highest affinity to pyrogallol (Km = 2.77 mM) followed by 4-methylcatechol (Km = 3.14 mM) and catechol (Km = 15.14 mM). The optimum pH for activity was 6.0. PPO was stable in the temperature range of 20-70 °C. PPO activity was completely inhibited by tropolone, ascorbic acid, sodium metabisulfite, and kojic acid at 0.1 mM. © 2014 American Chemical Society.

enzymatic browning; inhibitors; Manila mango (Mangifera indica); polyphenol oxidase; purification Purification; Biochemical characterization; Enzymatic browning; Mangifera indica; Polyphenol oxidase; Corrosion inhibitors; 4-methylcatechol; catechol; catechol derivative; catechol oxidase; enzyme inhibitor; pyrogallol; vegetable protein; antagonists and inhibitors; enzyme specificity; enzyme stability; enzymology; gel chromatography; ion exchange chromatography; isolation and purification; Mangifera; metabolism; molecular weight; pH; temperature; Catechol Oxidase; Catechols; Chromatography, Gel; Chromatography, Ion Exchange; Enzyme Inhibitors; Enzyme Stability; Hydrogen-Ion Concentration; Mangifera; Molecular Weight; Plant Proteins; Pyrogallol; Substrate Specificity; Temperature

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