title: Analysis of the activation of acetylcholinesterase by carbon nanoparticles using a monolithic immobilized enzyme microreactor: role of the water molecules in the active site gorge.
abstract: A biochromatographic system was used to study the direct effect of carbon nanoparticles (CNPs) on the acetylcholinesterase (AChE) activity. The AChE enzyme was covalently immobilized on a monolithic CIM-disk via its NH2 residues. Our results showed an increase in the AChE activity in presence of CNPs. The catalytic constant (k(cat)) was increased while the Michaelis constant (K(m)) was slightly decreased. This indicated an increase in the enzyme efficiency with increase of the substrate affinity to the active site. The thermodynamic data of the activation mechanism of the enzyme, i.e. ΔH* and ΔS*, showed no change in the substrate interaction mechanism with the anionic binding site. The increase of the enthalpy (ΔH*) and the entropy (ΔS*) with decrease in the free energy of activation (Ea) was related to structural conformation change in the active site gorge. This affected the stability of water molecules in the active site gorge and facilitated water displacement by substrate for entering to the active site of the enzyme.
authors.affiliation: UFR SMP , Besancon , France.
authors.forename: Firas
authors.initials: F
authors.lastname: Ibrahim
authors.forename: Claire
authors.initials: C
authors.lastname: Andre
authors.forename: Anne
authors.initials: A
authors.lastname: Iutzeler
authors.forename: Yves Claude
authors.initials: YC
authors.lastname: Guillaume
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry / Q000378:metabolism*; D019149:Bioreactors; D002244:Carbon / Q000737:chemistry*; D020134:Catalytic Domain; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D013816:Thermodynamics; D014867:Water / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D014867:Water; D002244:Carbon; D000110:Acetylcholinesterase