title: Purification and characterization of polyphenol oxidase from banana (Musa sapientum L.) pulp.
abstract: Polyphenol oxidase (EC 1.10.3.1, PPO) in the pulp of banana (Musa sapientum L.) was purified to 636-fold with a recovery of 3.0%, using dopamine as substrate. The purified enzyme exhibited a clear single band on polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS)-PAGE. The molecular weight of the enzyme was estimated to be about 41000 and 42000 by gel filtration and SDS-PAGE, respectively. The enzyme quickly oxidized dopamine, and its K(m) value for dopamine was 2.8 mM. The optimum pH was at 6.5, and the enzyme activity was stable in the range of pH 5-11 at 5 degrees C for 48 h. The enzyme had an optimum temperature of 30 degrees C and was stable even after a heat treatment at 70 degrees C for 30 min. The enzyme activity was completely inhibited by L-ascorbic acid, cysteine, sodium diethyldithiocarbamate, and potassium cyanide. Under a low buffer capacity, the enzyme was also strongly inhibited by citric acid and acetic acid at 10 mM.
authors.affiliation: Laboratory of Food Science, Faculty of Agriculture, Saga University, Saga 840-8502, Japan.
authors.forename: C P
authors.initials: CP
authors.lastname: Yang
authors.forename: S
authors.initials: S
authors.lastname: Fujita
authors.forename: M
authors.initials: M
authors.lastname: Ashrafuzzaman
authors.forename: N
authors.initials: N
authors.lastname: Nakamura
authors.forename: N
authors.initials: N
authors.lastname: Hayashi
mesh_terms: D004156:Catechol Oxidase / Q000737:chemistry / Q000302:isolation & purification*; D013379:Substrate Specificity; D019862:Zingiberales / Q000201:enzymology*
chemical_list: D004156:Catechol Oxidase