Herein, we report a facile technique to fabricate fluorescent polymeric structure, brilliant-green sequestered poly(amic) acid film (BG-PAA), as enzyme support-material for biosensor applications. The structure and fluorescence properties of the fabricated BG-PAA membrane were investigated using H-1 NMR, scanning electron microscopy (SEM) and fluorescence microscopy. Glucose oxidase (GOx) was used as a model enzyme that immobilized on BG-PAA membrane using glutaraldehyde. The created BGPAA/GOx membrane was utilized to fabricate dual-purpose as fluorescent and electrochemical biosensors for the determination of glucose in beverages. Utilization of the dissolved oxygen in GOx mediated glucose oxidation resulted in alteration of the measured fluorescence and current intensity. In the fluorescence assay, the decreased concentration of oxygen causes the increased fluorescence intensity due to the elimination of quenching effect of oxygen on BG fluorescence. In amperometric assay, decrease in oxygen concentration was followed at -0.7 V. After optimization of working conditions linearity, limit of detection and repeatability of system were determined. Finally, BG-PAA/GOx was tested to analyze glucose in beverages. (C) 2017 Elsevier B.V. All rights reserved.