Silica-immobilized His(6)-tagged enzyme: Alanine racemase in hydrophobic solvent

Silica-immobilized His(6)-tagged enzyme : Alanine racemase in hydrophobic solvent

QC 20100525 QC 20111128
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.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
Geobacillus stearothermophilus
enzyme surface
enzyme immobilization
His(6)-tag
bacillus-stearothermophilus
inhibition
resolution
esters

Ingår i:Biotechnology and Bioengineering: Wiley99:3, s. 712-7160006-35921097-0290

Av författaren/redakt...: Cassimjee, Karim ...; Trummer, Martin; Branneby, Cecili ...; Berglund, Per

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