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Fractional purification and kinetic parameters (Km and Vmax) of polyphenol oxidase extracted from three segments of Solanummelongenas and Musa sapientum fruits

P.C.Chikezie, A.R.Akuwudike, C.M.Chikezie, C.O.AndIbegbulem

Polyphenol oxidase (PPO) was extracted from three segments of Solanummelongenas andMusa sapientumfruits and partially purified. The specific activity of PPO was measured at each purification step to ascertain level of enzyme purity. In all cases, PPO conformed to Michaelis- Menten kinetics, showing different values of kinetics parameters.Michaelis- Menten constant for PPO (PPOKm) of S. melongenas mid-section and anterior segments showed no significant difference (p< 0.05), whereas the posterior gave PPOKm = 4.6±0.49 mM (p > 0.05).Maximum PPO activity (PPOVmax) was highest in the posterior segment: PPOVmax= 0.602±0.09 U. Mid-section of M. sapientum exhibited the highest Km value (PPOKm = 5.8±0.69 mM) compared with the anterior (PPOKm = 3.9±0.69 mM) (p> 0.05) and posterior PPOKm = 4.9±0.11mM segments (p< 0.05). Overall, M. sapientumPPOKm valueswere relatively higher than those of S.melongenas. Posterior S. melongenas exhibited the highest PPOVmax = 0.602±0.09 U, whereas the lowest value was registered in the anterior segment of M. sapientumPPOVmax = 0.234±0.09 U. Substrate specificity for PPO (PPOVmax/ Km) extracted from various segments of S. melongenas was in the increasing order of Mid-section > Posterior > Anterior, whereas that of M. sapientum wasMid-section >Anterior > Posterior. PPOVmax/Kmbetween the two fruits showed strong positive correlation (r = 0.862339). Catechol was a better substrate for PPOS. melongenas than PPOM. sapientum.The experimentally observed kinetic parameters of S. melongenas and M. sapientum signified the presence of PPO isoenzymes and non-uniform distribution of PPO in the two fruits.