Reversible sodium dodecyl sulfate activation of latent peach polyphenol oxidase by cyclodextrins.
The reversibility of the SDS-mediated activation of latent peach PPO has been studied using cyclodextrins as strip detergent agent. Cyclodextrins produced a combined inhibitory effect on enzymatic activity of latent peach PPO due to the complexation of detergent and the hydrophobic substrate 4-tert-butylcatechol (TBC) molecules. To study the reversibility of the activation process, this combined effect has to be separated. On the one hand, the enzyme was activated by acid-shocking and the activity was measured in the presence of cyclodextrins, using TBC as substrate. The inhibition curves obtained permitted study of the complexation of TBC into cyclodextrins. On the other hand, the enzyme was activated by SDS and the activity in the presence of cyclodextrins was measured using the highly hydrophilic o-diphenol dopamine as substrate. In this case, the inhibition curves obtained indicated the reversibility of the activation process when SDS was trapped by cyclodextrins. In addition, the complexation constant between SDS and 2-hydroxypropyl-beta-cyclodextrins was calculated by measuring conductivity (K(s) = 3500 M(-1)).
Laveda
F
F
Department of Biochemistry and Molecular Biology-A, University of Murcia, Campus de Espinardo, Murcia, E-30071, Spain.
Núñez-Delicado
E
E
García-Carmona
F
F
Sánchez-Ferrer
A
A
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D004156:Catechol Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002396:Catechols / Q000378:metabolism; D003505:Cyclodextrins / Q000494:pharmacology; D004298:Dopamine / Q000031:analogs & derivatives; D004553:Electric Conductivity; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005638:Fruit / Q000201:enzymology*; D007700:Kinetics; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D047392:beta-Cyclodextrins
D002396:Catechols; D003505:Cyclodextrins; D004791:Enzyme Inhibitors; D047392:beta-Cyclodextrins; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D012967:Sodium Dodecyl Sulfate; D004156:Catechol Oxidase; D004298:Dopamine