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Verlagsausgabe
DOI: 10.5445/IR/1000085375
Veröffentlicht am 23.08.2018
Originalveröffentlichung
DOI: 10.1016/j.apcata.2018.08.003

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.; Márquez-Álvarez, C.; Ferrer, M.

Abstract:
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 activitie ... mehr


Zugehörige Institution(en) am KIT Institut für Toxikologie und Genetik (ITG)
Institut für Technische Chemie und Polymerchemie (ITCP)
Publikationstyp Zeitschriftenaufsatz
Jahr 2018
Sprache Englisch
Identifikator ISSN: 0926-860X, 1873-3875
URN: urn:nbn:de:swb:90-853759
KITopen ID: 1000085375
Erschienen in Applied catalysis / A
Band 565
Seiten 59-67
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