Layer-by-layer films containing peptides of the Cry1Ab16 toxin from Bacillus thuringiensis for potential biotechnological applications

• Published in: Materials Science & Engineering C Vol. 61; pp. 832 - 841
• Main Authors: Plácido, Alexandra, de Oliveira Farias, Emanuel Airton, Marani, Mariela M., Vasconcelos, Andreanne G., Mafud, Ana C., Mascarenhas, Yvonne P., Eiras, Carla, Leite, José R.S.A., Delerue-Matos, Cristina
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2016
• Subjects: Alginates - chemistry; Atomic force microscopy; Bacillus thuringiensis; Bacillus thuringiensis - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Bioelectrochemistry; Chitosan - chemistry; Circular Dichroism; Cry1Ab16 toxin; Crystal structure; Electrochemical Techniques; Endotoxins - chemistry; Endotoxins - metabolism; Genetic modification; Genetically modified organism; Glucuronic Acid - chemistry; Hemolysin Proteins - chemistry; Hemolysin Proteins - metabolism; Hexuronic Acids - chemistry; Indicators; Layer-by-layer film; Microscopy, Atomic Force; Nanofibers - chemistry; Oxidation-Reduction; Peptides; Peptides - chemistry; Polyetherimides; Polyethyleneimine - chemistry; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared; Tin Compounds - chemistry; Toxins; Cry1Ab16 toxin; Layer-by-layer film; Genetically modified organism; Bioelectrochemistry
• ISSN: 0928-4931; 1873-0191
• DOI: 10.1016/j.msec.2016.01.011

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 from the 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 0.1mol·L−1 potassium phosphate buffer (PBS) at pH7.2. It was 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 200nm on the polymeric film. By UV–Visible spectroscopy (UV–Vis) it was observed that the ITO/PEI/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 system is an excellent candidate for applications in electrochemical sensors and other biotechnological applications for GMOs and environmental indicators. [Display omitted] •Peptides of the Cry1Ab16 toxin for potential biotechnological applications•Optimized LbL film deposition for synergic interactions of the conjugated materials•Characterization of self-assembled nanofibers with nanometric diameters•Future applications of LbL film in electrochemical sensors for GMOs