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October 2022

Automation of Enzyme Reactions: Ribonuclease

Journal/Book: Reprinted from ANALYTICAL BIOCHEMISTRY Volume 29 Number 3 June 1969 S. 515-525 Copyright (c) 1969 by Academic Press Inc. Printed in U. S. A.. 1968/1969;

Abstract: H. BARRERA K. S. CHIO AND A. L. TAPPEL Department of Food Science and Technology University of California Davis California 96616 Received November 29 1968 Automated techniques for studies of enzyme reactions have been developed recently. Stein et al. (1) reported the use of automated methodology to study the effects of cofactors and inhibitors on the reaction between ATP and brain microsomal adenosine triphosphatase. Methods for obtaining continuous enzyme data under gradient conditions were described by Tappel and Beck (2) in the determination of kinetic parameters of -glucuronidase. The AutoAnalyzer can be used for the study of timedependent reactions such as heat inactivation of enzymes and chemical inactivation (3). Mundry (4) described an automated method for the continuous determination of ribonuclease activity in effiuents 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 an ribonuclease activity. A sequentialsample 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 m of the dialyzable hydrolysis products of ribonucleic acid. ... schö

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