Autodisplay of an avidin with biotin-binding activity on the surface of Escherichia coli

• Published in: Biotechnology letters Vol. 40; no. 3; pp. 591 - 600
• Main Authors: Pardavé-Alejandre, H. D., Alvarado-Yaah, J. E., Pompa-Mera, E. N., Muñoz-Medina, J. E., Sárquiz-Martínez, B., Santacruz-Tinoco, C. E., Manning-Cela, R. G., Ortíz-Navarrete, V., López-Macías, C., González-Bonilla, C. R.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.03.2018; Springer Nature B.V
• Subjects: Amino acids; Applied Microbiology; Avidin; Bacteria; Binding; Biochemistry; Biomedical and Life Sciences; Biotechnology; Biotin; Coliforms; Confocal microscopy; Cytometry; E coli; Escherichia coli; Flow cytometry; Fluorescein; Fluorescence; Fluorescence microscopy; Fusion protein; Gel electrophoresis; Life Sciences; Microbiology; Microscopy; Original Research Paper; Ovalbumin; Proteins; Sodium lauryl sulfate; Autodisplay; surface display; ShdA; Biotin-binding activity; Avidin; In trans surface display
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-018-2507-6

Objectives To display a recombinant avidin fused to the autotransporter ShdA to bind biotinylated molecules on the surface of Escherichia coli . Results Two chimeric protein constructs containing avidin fused to the autotransporter ShdA were expressed on the surface of Escherichia coli DH5α. One fusion protein contained 476 amino acids of the ShdA α and β domains, whereas the second consisted of a 314 amino acid from α and truncated β domains. Protein production was verified by SDS-PAGE using an antibody to the molecular FLAG-tag. The surface display of the avidin-shdA fusion protein was confirmed by confocal microscopy and flow cytometry analysis, and the biotin-binding activity was evaluated by fluorescence microscopy and flow cytometry using biotin-4-fluorescein and biotinylated-ovalbumin (OVA). Conclusions Expression of a recombinant avidin with biotin-binding activity on the surface of E. coli was achieved using the autotransporter ShdA. This system is an alternative to bind biotinylated molecules to E. coli.