title: Electrogravimetric real-time and in situ michaelis-menten enzymatic kinetics: progress curve of acetylcholinesterase hydrolysis.
abstract: A piezoelectric detection of enzyme-modified surface was performed under Michaelis-Menten presumptions of steady-state condition. The approach herein presented showed promise in the study of enzymatic kinetics by measuring the frequency changes associated with mass changes at the piezoelectric crystal surface. Likewise, real-time frequency shifts, that is, dΔf/dt, indicated the rate of products formation from enzymatic reaction. In this paper, acetylcholinesterase was used as the enzymatic model and acetylcholine as substrate. The enzymatic rate has its maximum value for a short time during the kinetic reaction, for instance, during the first ten minutes of the reaction time scale. The values found for the kinetic constant rate and Michaelis-Menten constant were (1.4 ± 0.8) 10(5) s(-1) and (5.2 ± 3) 10(-4) M, respectively, in agreement with the values found in classical Michaelis-Menten kinetic experiments.
authors.affiliation: Instituto de Química, Universidade Estadual Paulista, CP 355, 14800-900, Araraquara, São Paulo, Brasil. prbueno@iq.unesp.br
authors.forename: Paulo R
authors.initials: PR
authors.lastname: Bueno
authors.forename: Ailton M
authors.initials: AM
authors.lastname: Watanabe
authors.forename: Ronaldo C
authors.initials: RC
authors.lastname: Faria
authors.forename: Márcio L
authors.initials: ML
authors.lastname: Santos
authors.forename: Carla S
authors.initials: CS
authors.lastname: Riccardi
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics
chemical_list: D000110:Acetylcholinesterase