title: Immobilization of enzymes on PTFE surfaces.
abstract: 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.
authors.affiliation: Institute for Pharmaceutical Biology, Nussallee 6, University of Bonn, 53115 Bonn, Germany. M.KEUSGEN@uni-bonn.de
authors.forename: M
authors.initials: M
authors.lastname: Keusgen
authors.forename: J
authors.initials: J
authors.lastname: Glodek
authors.forename: P
authors.initials: P
authors.lastname: Milka
authors.forename: I
authors.initials: I
authors.lastname: Krest
mesh_terms: D013437:Carbon-Sulfur Lyases / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D011138:Polytetrafluoroethylene
chemical_list: D004800:Enzymes, Immobilized; D011138:Polytetrafluoroethylene; D007770:L-Lactate Dehydrogenase; D013437:Carbon-Sulfur Lyases; C019243:alliin lyase