Author(s): Plácido, Alexandra ; Farias, Emanuel Airton de Oliveira ; Marani, Mariela M. ; Vasconcelos, Andreanne G. ; Mafud, Ana C. ; Mascarenhas, Yvonne P. ; Eiras, Carla ; Leite, José R.S.A. ; Delerue-Matos, Cristina
Date: 2016
Persistent ID: http://hdl.handle.net/10400.22/9213
Origin: Repositório Científico do Instituto Politécnico do Porto
Subject(s): Cry1Ab16 toxin; Genetically modified organism; Layer-by-layer film; Bioelectrochemistry
Cry1Ab16 is a toxin of crystalline insecticidal proteins that has been widely used in genetically modified organisms (GMOs) to gain resistance to pests. For the first time, in this study, peptides derived fromthe immunogenic Cry1Ab16 toxin (from Bacillus thuringiensis) were immobilized as layer-by-layer (LbL) films. Given the concern about food and environmental safety, a peptide with immunogenic potential, PcL342–354C, was selected for characterization of the electrochemical, optical, and morphological properties. The results obtained by cyclic voltammetry (CV) showed that the peptide have an irreversible oxidation process in electrolyte of 0.1 mol·L−1 potassiumphosphate buffer (PBS) at pH7.2. Itwas also observed that the electrochemical response of the peptide is governed mainly by charge transfer. In an attempt to maximize the electrochemical signal of peptide, it was intercalated with natural (agar, alginate and chitosan) or synthetic polymers (polyethylenimine (PEI) and poly(sodium 4-styrenesulfonate (PSS)). The presence of synthetic polymers on the film increased the electrochemical signal of PcL342–354C up to 100 times. Images by Atomic Force Microscopy (AFM) showed that the immobilized PcL342–354C formed self-assembled nanofibers with diameters ranging from 100 to 200 nm on the polymeric film. By UV–Visible spectroscopy (UV–Vis) it was observed that the ITO/PE/PSS/PcL342–354C film grows linearly up to the fifth layer, thereafter tending to saturation. X-ray diffraction confirmed the presence on the films of crystalline ITO and amorphous polypeptide phases. In general, the ITO/PEI/PSS/PcL342–354C film characterization proved that this systemis an excellent candidate for applications in electrochemical sensors and other biotechnological applications for GMOs and environmental indicators.
