title: Fast and easy enzyme immobilization by photoinitiated polymerization for efficient bioelectrochemical devices.
abstract: Immobilization and electrical wiring of enzymes is of particular importance for the elaboration of efficient biosensors and can be cumbersome. Here, we report a fast and easy protocol for enzyme immobilization, and as a proof of concept, we applied it to the immobilization of bilirubin oxidase, a labile enzyme. In the first step, bilirubin oxidase is mixed with a redox hydrogel "wiring" the enzyme reaction centers to electrodes. Then, this adduct is covered by an outer layer of PEGDA made by photoinitiated polymerization of poly(ethylene-glycol) diacrylate (PEGDA) and a photoclivable precursor, DAROCUR. This two-step protocol is 18 times faster than the current state-of-the-art protocol and leads to currents 25% higher. In addition, the outer layer of PEGDA acts as a protective layer increasing the lifetime of the electrode by 100% when operating continuously for 2000 s and by 60% when kept in dry state for 24 h. This new protocol is particularly appropriate for labile enzymes that quickly denaturate. In addition, by tuning the ratio PEGDA/DAROCUR, it is possible to make the enzyme electrodes even more active or more stable.
authors.affiliation: Université de Bordeaux, Centre de Recherche Paul Pascal, CNRS UPR 8641, 115 avenue Schweitzer, 33600 Pessac, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Suraniti
authors.forename: Vincent
authors.initials: V
authors.lastname: Studer
authors.forename: Neso
authors.initials: N
authors.lastname: Sojic
authors.forename: Nicolas
authors.initials: N
authors.lastname: Mano
mesh_terms: D004563:Electrochemistry / Q000295:instrumentation*; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007700:Kinetics; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000737:chemistry; D055668:Photochemical Processes; D011092:Polyethylene Glycols / Q000737:chemistry; D058105:Polymerization; D020072:Polyporales / Q000201:enzymology; D013997:Time Factors
chemical_list: D004800:Enzymes, Immobilized; C437167:poly(ethylene glycol)diacrylate; D011092:Polyethylene Glycols; D044925:Oxidoreductases Acting on CH-CH Group Donors; C042401:bilirubin oxidase