Tyrosinase kinetics: discrimination between two models to explain the oxidation mechanism of monophenol and diphenol substrates.
The kinetic behaviour of tyrosinase is very complex because the enzymatic oxidation of monophenol and o-diphenol to o-quinones occurs simultaneously with the coupled non-enzymatic reactions of the latter. Both reaction types are included in the kinetic mechanism proposed for tyrosinase (Mechanism I [J. Biol. Chem. 267 (1992) 3801-3810]). We previously confirmed the validity of the rate equations by the oxidation of numerous monophenols and o-diphenols catalysed by tyrosinase from different fruits and vegetables. Other authors have proposed a simplified reaction mechanism for tyrosinase (Mechanism II [Theor. Biol. 203 (2000) 1-12]), although without deducing the rate equations. In this paper, we report new experimental work that provides the lag period value, the steady-state rate, o-diphenol concentration released to the reaction medium. The contrast between these experimental data and the respective numerical simulations of both mechanisms demonstrates the feasibility of Mechanism I. The need for the steps omitted from Mechanism II to interpret the experimental data for tyrosinase, based on the rate equations previously deduced for Mechanism I is explained.
Fenoll
L G
LG
GENZ-Grupo de Investigación Enzimologi;a, Departamento de Bioquímica y Biología Molecular-A, Facultad de Biología, Universidad de Murcia, A. Correos 4021 E-30080, Murcia, Spain
Peñalver
M J
MJ
Rodríguez-López
J N
JN
Varón
R
R
García-Cánovas
F
F
Tudela
J
J
D000363:Agaricales / Q000201:enzymology*; D001671:Biochemistry / Q000379:methods*; D002849:Chromatography, Gas; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry; D007700:Kinetics; D013058:Mass Spectrometry; D008956:Models, Chemical; D014442:Monophenol Monooxygenase / Q000737:chemistry* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D019800:Phenol / Q000737:chemistry*; D013053:Spectrophotometry
D019800:Phenol; D014442:Monophenol Monooxygenase; D010100:Oxygen