Determination of intrinsic properties of immobilized enzymes
- 1 March 1981
- journal article
- Published by Springer Nature in Applied Biochemistry and Biotechnology
- Vol. 6 (1), 25-36
- https://doi.org/10.1007/bf02779684
Abstract
Staphylococcal nuclease has been insolubilized, directly through its amino groups, on CNBr-activated Sepharose 2B. For kinetic studies, a small substrate (thymidine 5′-(p-nitrophenyl phosphate) 3′-phosphate) has been used to measure the hydrolytic activity. With this system the absence of diffusional limitation has been proven. Eadie-Hofstee analysis of the data has been employed to determine the intrinsic kinetic constants of the insolubilized enzyme. Thek cat-pH andK M−pH profiles and the activation energies are similar for the soluble and for the insolubilized nuclease. At the same time conditions are established in which a stirred batch reactor containing particles of insolubilized nuclease behaves as an open system.Keywords
This publication has 11 references indexed in Scilit:
- Preparation of immobilized sepharose-micrococcal nuclease derivatives: activity and stabilityJournal of Solid-Phase Biochemistry, 1979
- Staphylococcal Nuclease Immobilized on Agarose GelPublished by Springer Nature ,1978
- [29] Kinetic behavior of immobilized enzyme systemsMethods in Enzymology, 1976
- Preparation of cyanogen bromide-activated agarose gelsJournal of Chromatography A, 1973
- Effect of internal diffusion in heterogeneous enzyme systems: Evaluation of true kinetic parameters and substrate diffusivityJournal of Theoretical Biology, 1973
- The pH Dependence of the Steady State Kinetic Parameters for Staphylococcal Nuclease-catalyzed Hydrolysis of Deoxythymidine-3′-phosphate-5′-p-nitrophenylphosphate in H2O and D2OJournal of Biological Chemistry, 1973
- Regulation of Escherichia coli phosphofructokinase in situBiochemical and Biophysical Research Communications, 1973
- Kinetic behavior at high enzyme concentrations. Magnitude of errors of Michelis-Menten and other approximations.1970
- Action of staphylococcal nuclease on synthetic substratesBiochemistry, 1969
- Catalytic Properties and Specificity of the Extracellular Nuclease of Staphylococcus aureusJournal of Biological Chemistry, 1967