title: Kinetic study of hydroxytyrosol oxidation and its related compounds by Red Globe grape polyphenol oxidase.
abstract: Red Globe grape polyphenol oxidase, partially purified using phase partitioning with Triton-X114, was used to study the oxidation of hydroxytytosol (HT) and its related compounds tyrosol (TS), tyrosol acetate (TSA), and hydroxytyrosol acetate (HTA). The enzyme showed activity toward both monophenols (monophenolase activity) and o-diphenols (diphenolase activity) with a pH optimum (pH 6.5) that was independent of the phenol used. However, the optimal temperature for diphenolase activity was substrate-dependent, with a broad optimum of 25-65 °C for HT, compared with the maximum obtained for HTA (40 °C). Monophenolase activity showed the typical lag period, which was modulated by pH, substrate and enzyme concentrations, and the presence of catalytic amounts of o-diphenols. When the catalytic power (Vmax/K(M)) was determined for both activities, higher values were observed for o-diphenols than for monophenols: 9-fold higher for the HT/TS pair and 4-fold higher for HTA/TSA pair. Surprisingly, this ratio was equally higher for TSA (2.2-fold) compared with that of TS, whereas no such effect was observed for o-diphenols. This higher efficiency of TSA could be related to its greater hydrophobicity. Acetyl modification of these phenols not only changes the kinetic parameters of the enzyme but also affects their antioxidant activity (ORAC-FL assays), which is lower in HTA than in HT.
authors.affiliation: Department of Biochemistry and Molecular Biology-A, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia , Campus Espinardo, E-30100 Murcia, Spain.
authors.forename: María Inmaculada
authors.initials: MI
authors.lastname: García-García
authors.forename: Samanta
authors.initials: S
authors.lastname: Hernández-García
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Sánchez-Ferrer
authors.forename: Francisco
authors.initials: F
authors.lastname: García-Carmona
mesh_terms: D004156:Catechol Oxidase / Q000737:chemistry*; D007700:Kinetics; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010626:Phenylethyl Alcohol / Q000031:analogs & derivatives* / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry*; D027843:Vitis / Q000737:chemistry / Q000201:enzymology*
chemical_list: D010940:Plant Proteins; C005975:3,4-dihydroxyphenylethanol; D004156:Catechol Oxidase; D010626:Phenylethyl Alcohol