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• W1976356985 endingPage "10230" @default.
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• W1976356985 abstract "A partial characterization of polyphenol oxidase (PPO) activity in Ocimum basilicum L. is described. PPO in O. basilicum L. was extracted and purified through (NH4)2SO4 precipitation, dialysis, and a Sepharose 4B−l-tyrosine-p-aminobenzoic acid affinity column. The samples obtained from (NH4)2SO4 precipitation and dialysis were used for the characterization of PPO. At the end of purification by affinity chromatography, 11.5-fold purification was achived. The purified enzyme exhibited a clear single band on sodium dodecyl sulfate−polyacrylamide gel electrophoresis. The molecular mass of the enzyme was estimated to be ∼54 kDa. The contents of total phenolic and protein of O. basilicum L. extracts were determined. The total phenolic content of O. basilicum L. was determined spectrophotometrically according to the Folin−Ciocalteu procedure and was found to be 280 mg 100 g-1 on a fresh weight basis. The protein content was determined according to the Bradford method. The enzyme showed activity to 4-methylcatechol, catechol, and pyrogallol substrates, but not to tyrosine. Therefore, of these three substrates, 4-methylcatecol was the best substrate due to the highest Vmax/Km value, followed by pyrogallol and catechol. The optimum pH was at 6, 8, and 9 for 4-methylcatechol, catechol, and pyrogallol, respectively. The enzyme had an optimum temperature of 20, 40, and 50 °C for 4-methylcatechol, catechol, and pyrogallol, respectively. It was found that optimum temperature and pH were dependent on the substrates studied. The enzyme activity with increasing temperature and inactivation time for 4-methylcatechol, catechol, and pyrogallol substrates decreased due to heat denaturation of the enzyme. Keywords: Polyphenol oxidase; Ocimum basilicum L.; purification; substrate specificity; pH; temperature; heat inactivation" @default.
• W1976356985 created "2016-06-24" @default.
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• W1976356985 date "2005-11-23" @default.
• W1976356985 modified "2023-10-17" @default.
• W1976356985 title "Purification and Characterization of <i>Ocimum basilicum</i> L. Polyphenol Oxidase" @default.
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• W1976356985 doi "https://doi.org/10.1021/jf051646j" @default.
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