Proteolytic activation of latent Paraguaya peach PPO. Characterization of monophenolase activity.
The kinetics of the activation process of latent peach PPO by trypsin was studied. By coupling this activation process to the oxidation of 4-tert-butylcatechol (TBC) to its corresponding quinone, it was possible to evaluate the specific rate constant of active PPO formation, k(3), which showed a value of 0.04 s(-1). This proteolytic activation of latent peach PPO permitted us to characterize the monophenolase activity of peach PPO for the first time using p-cresol as substrate, and it showed the characteristic lag period of the kinetic mechanism of monophenols hydroxylation, which depended on the enzyme and substrate concentration, the pH and the presence of catalytic amounts of o-diphenol (4-methylcatechol). The enzyme activation constant, k(act), was 2 microM.
Department of Biochemistry and Molecular Biology-A, Faculty of Biology, University of Murcia, Campus de Espinardo, E-30071 Murcia, Spain.
F
F
Laveda
E
E
Núñez-Delicado
F
F
García-Carmona
A
A
Sánchez-Ferrer
D004156:Catechol Oxidase / Q000378:metabolism*; D002396:Catechols / Q000378:metabolism; D004789:Enzyme Activation; D005638:Fruit / Q000201:enzymology*; D007700:Kinetics; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D002396:Catechols; C038096:tert-butylcatechol; D010088:Oxidoreductases; D004156:Catechol Oxidase; C412191:monophenolase; D014357:Trypsin