Publication
Title
Active site titration of immobilized beta-galactosidase for the determination of active enzymes
Author
Abstract
In the present study, an active site titration method is demonstrated, to determine the amount of active enzyme (beta-galactosidase), immobilized on a support. Two types of supports were investigated, viz, amino acrylic resin and a mixed matrix membrane. Furthermore, 2',4'-dinitrophenyl 2-deoxy-2-fluoro-beta-D-galactopyranoside was used as an inhibitor for the active site titration of immobilized beta-galactosidase obtained from Kluyveromyces lactis. Using the active site titration, approximately 8.3 mg of active enzyme was found on 1 g of dried commercially available SPRIN imibond, which is an amino acrylic resin with covalently bound beta-galactosidase obtained from K. lactis. However, this method, in its present form, was not effective on the mixed matrix membranes due to the irreversible partial adsorption of the leaving group (2',4'-dinitrophenolate) by the membrane. This observation implied that it is important to investigate interactions between the support and the used inhibitor and leaving group. (C) 2014 Elsevier B.V. All rights reserved.
Language
English
Source (journal)
Biochemical engineering journal. - Amsterdam, 1998, currens
Publication
Amsterdam : Elsevier , 2015
ISSN
1369-703X [print]
1873-295X [online]
DOI
10.1016/J.BEJ.2014.10.007
Volume/pages
93 (2015) , p. 137-141
ISI
000347362100018
Full text (Publisher's DOI)
Full text (publisher's version - intranet only)
UAntwerpen
Faculty/Department
Research group
Publication type
Subject
Affiliation
Publications with a UAntwerp address
External links
Web of Science
Record
Identifier
Creation 13.03.2015
Last edited 09.10.2023
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