Bounded water kinetic model of beta-galactosidase in reverse micelles.
In the present investigation, beta-galactosidase was solubilized into Aerosol OT (AOT)/isooctane reverse micelles. Kinetic data for the hydrolysis of o-nitrophenyl-beta-D-galactopyranoside (ONPG) at different pH values and molar ratios of water to AOT (Wo) were collected. It was observed that the usual kinetic model used for beta-galactosidase catalysis in aqueous systems failed to represent the experimental data. A bounded water model, however, showed a better correlation between enzymatic activity and Wo. In contrast to the aqueous system, controlling the water concentration in the reverse micelles allows the rate constants for the reaction between water molecules and glycosyl-enzyme complexes to be evaluated.
Chen
C Will
CW
Department of Bioengineering, Tatung University, 40 Chungshan Road, 3rd Section, Taipei, Taiwan, Republic of China. Will@ttu.edu.tw
Ou-Yang
Chao-Chih
CC
D002384:Catalysis; D003102:Colloids / Q000737:chemistry*; D003198:Computer Simulation; D004143:Dioctyl Sulfosuccinic Acid / Q000737:chemistry*; D004789:Enzyme Activation; D007700:Kinetics; D008823:Micelles; D008956:Models, Chemical; D009795:Octanes / Q000737:chemistry; D011485:Protein Binding; D014867:Water / Q000737:chemistry*; D001616:beta-Galactosidase / Q000737:chemistry*
D003102:Colloids; D008823:Micelles; D009795:Octanes; D014867:Water; D004143:Dioctyl Sulfosuccinic Acid; D001616:beta-Galactosidase; C045798:2,2,4-trimethylpentane