Silica-immobilized His(6)-tagged enzyme: Alanine racemase in hydrophobic solvent
2008 (English)In: Biotechnology and Bioengineering, ISSN 0006-3592, E-ISSN 1097-0290, Vol. 99, no 3, 712-716 p.Article in journal (Refereed) Published
A new immobilization method for enzymes is presented to facilitate synthetic applications in aqueous as well as organic media. The enzyme Alanine racemase (AlaR) from Geobacillus stearothermophilus was cloned, overexpressed and then immobilized on a silica-coated thin-layer chromatography plate to create an enzyme surface. The enzyme, fused to a His(6)-tag at its N-terminal, was tethered to the chemically modified silica-coated TLC plate through cobalt ions. The immobilized enzyme showed unaltered kinetic parameters in small-scale stirred reactions and retained its activity after rinsing, drying, freezing or immersion in n-hexane. This practical method is a first step towards a general immobilization method for synthesis applications with any enzyme suitable for His(6)-tagging.
Place, publisher, year, edition, pages
2008. Vol. 99, no 3, 712-716 p.
Geobacillus stearothermophilus, enzyme surface, enzyme immobilization, His(6)-tag, bacillus-stearothermophilus, inhibition, resolution, esters
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-17276DOI: 10.1002/bit.21587ISI: 000252511800023PubMedID: 17680683ScopusID: 2-s2.0-38449117584OAI: oai:DiVA.org:kth-17276DiVA: diva2:335319
QC 20100525 QC 201111282010-08-052010-08-052011-11-28Bibliographically approved