Electrochemical Polymerization of (2-Dodecyl-4, 7-di (thiophen-2-yl)-2H-benzo[d][1,2,3] triazole): A Novel Matrix for Biomolecule Immobilization

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dc.contributor.authorEkiz F.
dc.contributor.authorYuksel M.
dc.contributor.authorBalan A.
dc.contributor.authorTimur S.
dc.contributor.authorToppare L.
dc.date.accessioned2019-10-27T08:34:19Z
dc.date.available2019-10-27T08:34:19Z
dc.date.issued2010
dc.departmentEge Üniversitesien_US
dc.description.abstractA recently synthesized conducting polymer [poly(2-dodecyl-4,7-di(thiophen-2-yl)-2H-benzo[d][1,2,3]triazole (PTBT)] was tested as a platform for biomolecule immobilization. After electrochemical polymerization of the monomer (TBT) on graphite electrodes, immobilization of glucose oxidase (GOx, ß-D-glucose: oxygen-1-oxidoreductase, EC 1.1.3.4) was carried out. To improve the interactions between the enzyme and hydrophobic alkyl chain on the polymeric structure, GOx and isoleucine (Ile) amino acid were mixed in sodium phosphate buffer (pH 7.0) with a high ionic strength (250 × 10-3 M). The solution is then casted on the polymer film, and the amino groups in the protein structure were crosslinked using glutaraldehyde (GA) as the bifunctional agent. Finally, the surface was covered with a perm-selective membrane. Consequently, cross-linked enzyme crystal (CLEC) like assembles with regular shapes were observed after immobilization. Microscopic techniques such as scanning electron microscopy (SEM) and fluorescence microscopy were used to monitor the surface morphologies of both the polymer and the bioactive layer. Electrochemical responses of the enzyme electrodes were measured by monitoring O2 consumption in the presence of glucose at -0.7 V. The optimized biosensor showed a very good linearity between 0.05 and 2.5 × 10-3 M with a 52 s response time and a detection limit (LOD) of 0.029 × 10-3 M to glucose. Also, kinetic parameters, operational and storage stabilities were determined. Km and Imax values were found as 4.6 × 10-3 M and 2.49 µA, respectively. It was also shown that no activity was lost during operational and storage conditions. Finally, proposed system was applied for glucose biomonitoring during fermentation in yeast culture where HPLC was used as the reference method to verify the data obtained by the proposed biosensor. A newly synthesized conducting polymer [poly(2-dodecyl-4,7-di(thiophen-2-yl)-2H-benzo[d][1,2,3]triazole (PTBT)] as a matrix for biomolecule immobilization was reported here. Glucose oxidase (GOx) was used as the model enzyme to examine the possibility of its immobilization onto PTBT after electrochemical polymerization of TBT on the graphite surface. Microscopic techniques such as scanning electron microscopy (SEM) and fluorescence microscopy were used to monitor the surface morphologies of both the polymer and the bioactive layer. After optimization and characterization studies, designed biosensor was applied for glucose monitoring in yeast culture. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.en_US
dc.identifier.doi10.1002/mabi.201000185en_US
dc.identifier.endpage1565en_US
dc.identifier.issn1616-5187
dc.identifier.issue12en_US
dc.identifier.pmid20957699en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage1557en_US
dc.identifier.urihttps://doi.org/10.1002/mabi.201000185
dc.identifier.urihttps://hdl.handle.net/11454/27018
dc.identifier.volume10en_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.relation.ispartofMacromolecular Bioscienceen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectCLEC (cross-linked enzyme crystal) structureen_US
dc.subjectConducting polymersen_US
dc.subjectElectrochemical polymerizationen_US
dc.subjectEnzyme biosensorsen_US
dc.subjectHydrophobic interactionsen_US
dc.titleElectrochemical Polymerization of (2-Dodecyl-4, 7-di (thiophen-2-yl)-2H-benzo[d][1,2,3] triazole): A Novel Matrix for Biomolecule Immobilizationen_US
dc.typeArticleen_US

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