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Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

Coscolín, C.; Beloqui, A.; Martínez-Martínez, M.; Bargiela, R.; Santiago, G.; Blanco, R. M.; Delaittre, G. 1,2; Márquez-Álvarez, C.; Ferrer, M.
1 Institut für Toxikologie und Genetik (ITG), Karlsruher Institut für Technologie (KIT)
2 Institut für Technische Chemie und Polymerchemie (ITCP), Karlsruher Institut für Technologie (KIT)


It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg$^{-1}$ for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH$_{2}$-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg$^{-1}$. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g$^{-1}$ and specific activities of up to 29.9 ± 0.3 U mg$^{-1}$, were obtained. ... mehr

Verlagsausgabe §
DOI: 10.5445/IR/1000085375
Veröffentlicht am 23.08.2018
DOI: 10.1016/j.apcata.2018.08.003
Zitationen: 20
Web of Science
Zitationen: 17
Zitationen: 22
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Technische Chemie und Polymerchemie (ITCP)
Institut für Toxikologie und Genetik (ITG)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2018
Sprache Englisch
Identifikator ISSN: 0926-860X, 1873-3875
KITopen-ID: 1000085375
HGF-Programm 47.01.01 (POF III, LK 01) Biol.Netzwerke u.Synth.Regulat. ITG+ITC
Erschienen in Applied catalysis / A
Verlag Elsevier
Band 565
Seiten 59-67
Vorab online veröffentlicht am 06.08.2018
Nachgewiesen in Web of Science
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