Advantages of continuous over batch reactors for the kinetic analysis of enzymes inhibited by an unknown substrate impurity.
A new experimental technique, employing a continuous stirred-tank reactor, for studying enzyme kinetics in the presence of inhibitor-contaminated substrate is described. The proposed method is simulated mathematically for competitive, uncompetitive, and mixed-type noncompetitive inhibition. The step-by-step experimental procedure is described, as is the necessary data analysis for determining the kinetic parameters. Differences in system response for enzyme inhibition by excess substrate and by an impurity are illustrated, and a stability analysis of the system is performed.
Gallifuoco
Alberto
A
Dipartimento di Chimica, Ingegneria Chimica e Materiali, Università di L'Aquila, 67040 Monteluco di Roio, AQ, Italy. gallifuo@ing.univaq.it
Alfani
Francesco
F
Cantarella
Maria
M
D019149:Bioreactors; D003198:Computer Simulation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000378:metabolism*; D004798:Enzymes / Q000378:metabolism*; D008954:Models, Biological; D008956:Models, Chemical; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D004798:Enzymes