Binding of cetylpyridinum chloride to glucose oxidase

The binding of cetylpyridinum chloride (CPC) with glucose oxidase (GOD) has been extensively studied at various experimental conditions such as ionic strength, urea concentration and pH at 25 degrees C, using ion-selective membrane electrodes, UV-vis absorption spectroscopy and enzyme activity assay method. The accurate binding isotherms have been obtained and analyzed in terms of Scatchard plot and binding capacity concept. The results represent two binding set system for most of studied conditions. The values of Hill equation parameters have been estimated and used for calculation of intrinsic Gibbs free energy of binding. The results have been interpreted in terms of structural viewpoint of GOD and nature of interactions in the solution. The interpretations are in good agreement with denaturation experiment. Activity measurements represent the significant activation of enzyme due to binding of first CPC molecules. However, the binding of subsequent CPC diminished the activity of enzyme which may be due to the binding of second CPC to enzyme active site. The complete deactivation of enzyme is reached due to binding of about five CPC ions.