Title
Active site titration of immobilized beta-galactosidase for the determination of active enzymes Active site titration of immobilized beta-galactosidase for the determination of active enzymes
Author
Faculty/Department
Faculty of Sciences. Bioscience Engineering
Publication type
article
Publication
Amsterdam ,
Subject
Biology
Engineering sciences. Technology
Source (journal)
Biochemical engineering journal. - Amsterdam
Volume/pages
93(2015) , p. 137-141
ISSN
1369-703X
ISI
000347362100018
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
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.
E-info
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