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Optimization of enzyme immobilization on magnetic microparticles using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as a crosslinking agent

Kazenwadel, F.; Wagner, H.; Rapp, B.E.; Franzreb, M.

Enzyme immobilization is a versatile tool in biotransformation processes to enhance enzyme activity and to secure an easy separation of catalysts and products and the reusability of enzymes. A simple and commonly used method for crosslinking enzymes to a solid support is the zero-length crosslinking agent 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). This work shows the optimization of the EDC-crosslinking protocol for two enzymes, glucose oxidase (GOx) and horseradish peroxidase (HRP), to functionalized magnetic microparticles. For GOx the optimization of the immobilization parameters pH-value and the enzyme to particle ratio results in activity yields of up to 36%, which is in the usual range for undirected enzyme immobilisations. In contrast, for HRP the activity yield does not exceed 6% even after optimization of the protocols. The main reasons for this unusually low activity yield are the presence of multiple HRP isoforms in the enzyme solution used for immobilisation and the observed tendency of HRP to be inactive even in the case of simple physisorption to the particle surface. © 2015 The Royal Society of Chemistry.

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DOI: 10.5445/IR/1000052005
DOI: 10.1039/c5ay02670a
Zitationen: 14
Web of Science
Zitationen: 13
Zugehörige Institution(en) am KIT Institut für Mikrostrukturtechnik (IMT)
Institut für Funktionelle Grenzflächen (IFG)
Publikationstyp Zeitschriftenaufsatz
Jahr 2015
Sprache Englisch
Identifikator ISSN: 1759-9660
KITopen-ID: 1000052005
HGF-Programm 47.02.06 (POF III, LK 01)
Erschienen in Analytical Methods
Band 7
Heft 24
Seiten 10291-10298
Bemerkung zur Veröffentlichung RSC geförderter Open Access-Artikel
Nachgewiesen in Scopus
Web of Science
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