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
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