Molecular characterization of the enhancing immunoglobulin of mice. Abstr.

Abstract

The α-chymotrypsin-catalyzed hydrolysis of the p-nitrophenyl acetate in the solvent mixtures containing from 1.6 up to 10% (v/v) acetonitrile in the presence of aqueous Tris buffer at pH 8.0 was investigated at 298 K by use of an isoperibolic batch calorimeter. A special experimental arrangement of the reaction components for the investigation of the hydrolytically instable substrate was used. Furthermore, the release of p-nitrophenol was recorded with an UV-vis-spectrophotometer under comparable conditions. The calorimetric curves consist of two parts. The first part is strongly rising and finished by a break point in the ΔT (time) curve. This first step is dominated by the enzyme-catalyzed reaction. After the break point a slow non-enzymatic process determines the course of the calorimetric curve. The molar enthalpy changes of overall reaction (ester hydrolysis and buffer protonation) of -100 ± 8, -106 ± 5 and -102 ± 5 kJ/mol were evaluated by a combination of the results from the spectrophotometric and calorimetric data for 1.6, 4.0 and 10.0% acetonitrile mixtures, respectively. The obtained results indicate that the enzyme-catalyzed hydrolysis is suitable for quantitative determination of the hydrophobic ester p-nitrophenyl acetate in water-acetonitrile mixtures using calorimetric detection. © 2004 Elsevier B.V. All rights reserved