Glutathione transferases immobilized on nanoporous alumina: Flow system kinetics, screening, and stability
2014 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 446, 59-63 p.Article in journal (Refereed) Published
The previously uncharacterized Drosophila melanogaster Epsilon-class glutathione transferases E6 and E7 were immobilized on nanoporous alumina. The nanoporous anodized alumina membranes were derivatized with 3-aminopropyl-triethoxysilane, and the amino groups were activated with carbonyldiimidazole to allow coupling of the enzymes via c-amino groups. Kinetic analyses of the immobilized enzymes were carried out in a circulating flow system using CDNB (1-chloro-2,4-dinitrobenzene) as substrate, followed by specificity screening with alternative substrates. A good correlation was observed between the substrate screening data for immobilized enzyme and corresponding data for the enzyme in solution. A limited kinetic study was also carried out on immobilized human GST S1-1 (also known as hematopoietic prostaglandin D synthase). The stability of the immobilized enzymes was virtually identical to that of enzymes in solution, and no leakage of enzyme from the matrix could be observed.
Place, publisher, year, edition, pages
2014. Vol. 446, 59-63 p.
Immobilization, Glutathione transferase, Kinetics, Screening, Enzyme reactor, Drosophila
Biochemistry and Molecular Biology Analytical Chemistry
Research subject Neurochemistry with Molecular Neurobiology
IdentifiersURN: urn:nbn:se:su:diva-100853DOI: 10.1016/j.ab.2013.10.004ISI: 000329949500010OAI: oai:DiVA.org:su-100853DiVA: diva2:698344