Immobilization of enzymes on PTFE surfaces
| Autor: | Peter Milka, Ingo Krest, Michael Keusgen, Janina Glodek |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
biology
Immobilized enzyme L-Lactate Dehydrogenase Bioengineering Enzymes Immobilized Applied Microbiology and Biotechnology Combinatorial chemistry Benzoquinone chemistry.chemical_compound Carbon-Sulfur Lyases Membrane chemistry Covalent bond Concanavalin A Alliinase biology.protein Organic chemistry Bifunctional Biosensor Polytetrafluoroethylene Biotechnology |
| Zdroj: | Biotechnology and bioengineering. 72(5) |
| ISSN: | 0006-3592 |
| Popis: | Membranes and powders prepared from PTFE (polytetrafluorethylene) were investigated for their potential use as multifunctional supports for enzymes. The obtained bioactive materials are valuable for the construction of biosensors and enzyme reactors. To allow covalent coupling of enzymes to PTFE, the surface of the material was treated with elementary sodium followed by oxidation with ozone or hydrogen peroxide.%Derivatization steps were optimized in order to achieve highest enzyme loading and short reaction times. Alliinase (EC 4.4.1.4) and L-lactic dehydrogenase (EC 1.1.1.27) were chosen as model enzymes and were either immobilized by covalent coupling or fixed indirectly by a sugar-lectin binding. For the latter method, the sugar mannan was bound to the membrane surface as an anchor for layers of the lectin concanavalin A and the alliinase. Highest alliinase loading was achieved at 0.2 microg x cm(-2). Immobilization of alliinase via the lectin concanavalin A and a bifunctional epoxide gave the best long-term stability.%L-Lactic dehydrogenase was most sufficiently immobilized by using benzoquinone as spacer. These procedures show several advantages: 1) enzymes can be immobilized under physiological conditions, 2) an enzyme-multilayer can be achieved, and 3) protein layers are renewable. |
| Databáze: | OpenAIRE |
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