AUTOMATION OF ENZYME REACTIONS - RIBONUCLEASE

Automated techniques for studies of enzyme reaction have been developed recently. Stein et al. (1) reported the use of automated meethodology to study the effects of cofactors and inhibitions on the reaction between ATP and brain microsomal adenosine triphosphatase. Methods for obtaining continous enzyme data under gradient conditions were described by Tappel and Beck (2) in the determination of kinetic parameters of β-glucironidase. The AutoAnalyzer can be used for the study of time-dependent reactions, such as heat inactivation of enzymes and chemical inactivation (3). Mundry (4) described an automated method of chromatographic columns. This paper describes the application of a linear gradient device and techniques to the detailed studies of effects of enzyme and substrate concentrations, pH, and temperature on ribonuclease activity . A sequential-sample method for determination of the enzyme activity at a sampling rate of 20 samples per hour is also described. These methods are based on the quantitative determination of absorbance at 260 mμ of the dialyzable hydrolysis products of ribonucleic acid. © 1969.