The influence of the support nature on the kinetics parameters, inhibition constants and reactivation of immobilized acetylcholinesterase

Acetylcholinesterase (AChE) was immobilized on two different composite membranes constituted by a chemically modified poly-acrylonitrile (PAN) membrane plus a layer of tethered chitosan of different molecular weight, 10 kDa or 400 kDa. AChE was also directly immobilized on a chemically modified PAN membrane with NaOH and ethylenediamine (EDA) without chitosan. To know how the different supports affected the enzyme activity and the kinetic parameters, the AChE activity was studied in the soluble form and in the insoluble form with all the three types of modified PAN membranes. The best performance was obtained by the modified PAN membrane having the chitosan with the lower molecular weight.The results concerning the AChE inhibition by methyl-paraoxon and the subsequent reactivation by pyridine-2-aldoxime methochloride (2-PAM) are presented and discussed. The composite membrane having chitosan with the lower molecular weight appeared to be potentially useful for applications in the field of biosensors.