simple graphical method for the determination of reversible inhibition type,inhibitio constant(Ki) and estimation of fifty per cent of inhibition I50 of an enzyme reaction is described.The method consists of plotting experimental data as “degree ofinhibition” versus the inhibitor concentration at two or more concentrations of substrate.Diagnosis of inhibition type is based on determination of I50 and the observation of the shift of the inhibition curves.Relationship between I50 and inhibition constant Ki was discussed. As implified hyperbolae equation of degree of inhibition showing kinetic orders of 1 and zero at low and high concentrations of inhibitors respectively is proposed.The relative error of inhibitor concentration increased drastically when degree of inhibition reached values of 90%.Examples of published inhibition reports as well as an experimental example of amperometric biosensor based on tyrosinase inhibition by benzoic acid were in agreemen with the proposed theoretical approach.
Amine, A., El Harrad, L., Arduini, F., MOSCONE DINIA, D., Palleschi, G. (2014). Analytical aspects of enzyme reversible inhibition. TALANTA, 118, 368-374 [10.1016/j.talanta.2013.10.025].
Analytical aspects of enzyme reversible inhibition
ARDUINI, FABIANA;MOSCONE DINIA, DANILA;PALLESCHI, GIUSEPPE
2014-01-01
Abstract
simple graphical method for the determination of reversible inhibition type,inhibitio constant(Ki) and estimation of fifty per cent of inhibition I50 of an enzyme reaction is described.The method consists of plotting experimental data as “degree ofinhibition” versus the inhibitor concentration at two or more concentrations of substrate.Diagnosis of inhibition type is based on determination of I50 and the observation of the shift of the inhibition curves.Relationship between I50 and inhibition constant Ki was discussed. As implified hyperbolae equation of degree of inhibition showing kinetic orders of 1 and zero at low and high concentrations of inhibitors respectively is proposed.The relative error of inhibitor concentration increased drastically when degree of inhibition reached values of 90%.Examples of published inhibition reports as well as an experimental example of amperometric biosensor based on tyrosinase inhibition by benzoic acid were in agreemen with the proposed theoretical approach.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
