Partial purification of latent polyphenol oxidase from peach (Prunus persica L. Cv. Catherina). Molecular properties and kinetic characterization of soluble and membrane-bound forms.
This paper analyzes the kinetic and structural characteristics of polyphenol oxidase (PPO) from peach cv. Catherina. The PPO was obtained in a latent state in both the soluble and membrane-bound forms, and both forms were activated by acid shock and the detergent SDS. Plant defense is the main function assigned to PPO, which would be activated by the acid environment resulting from tissue damage. On the other hand, it has been suggested that, physiologically, the role played by SDS may be fulfilled by lipids. Native isoelectric focusing identified two acid isoforms of pI 5.7 and 5.8 for the soluble form and one isoform with pI 5.7 for the membrane-bound form. A partially denaturing SDS-PAGE revealed two very close bands of activity in both cases, but the Western blot performed on a totally denaturing SDS-PAGE, using polyclonal antibodies against bean PPO, revealed a single band in the membrane-bound fraction with a molecular mass of 60 kDa.
Cabanes
Juana
J
Departamento de Bioquímica y Biología Molecular A, Facultad de Biología, Universidad de Murcia, Campus de Espinardo, CP 30100 Murcia, Spain. jcabanes@um.es
Escribano
Josefa
J
Gandía-Herrero
Fernando
F
García-Carmona
Francisco
F
Jiménez-Atiénzar
Mercedes
M
D004156:Catechol Oxidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005638:Fruit / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D027861:Prunus / Q000201:enzymology*; D012995:Solubility
D004156:Catechol Oxidase