title: Development of a whole-cell biosensor by cell surface display of a gold-binding polypeptide on the gold surface.
abstract: Cell surface display was used as a strategy to display the gold-binding polypeptide (GBP) fusion protein on the surface of Escherichia coli, and consequently to immobilize the cells on the gold surface. The DNA encoding the GBP was fused to the truncated fadL gene and was expressed by the tac promoter. For the display of the core streptavidin (cSA) of Streptomyces avidinii, the cSA gene was fused to the truncated oprF gene. After the dual display of FadL-GBP and OprF-cSA on the surface of E. coli, binding of cells on the gold surface and the interaction of OprF-cSA with the biotin-horseradish peroxidase (HRP) were studied by surface plasmon resonance (SPR) analysis. Cells displaying the FadL-GBP fusion protein could be immobilized on the SPR sensor chip as shown by the SPR angle shift of 0.5 degrees , which was stably bound at least for 60 h with a washing solution. When the FadL-GBP and OprF-cSA fusion proteins were displayed on the same cell surface, the former was used to immobilize the cells on the gold surface and the latter was used for the interaction studies with the biotin-HRP, which demonstrates that the strategy should be useful for developing whole-cell biosensor chips.
authors.affiliation: BioProcess Engineering Research Center, Center for Systems and Synthetic Biotechnology, Institute for the BioCentury, KAIST, Daejeon, Korea.
authors.forename: Tae Jung
authors.initials: TJ
authors.lastname: Park
authors.forename: Shun
authors.initials: S
authors.lastname: Zheng
authors.forename: Yeon Jae
authors.initials: YJ
authors.lastname: Kang
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism; D018914:Cells, Immobilized; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism*; D006046:Gold / Q000737:chemistry / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D006046:Gold; D006735:Horseradish Peroxidase