title: Biocatalytic Formation of Gold Nanoparticles Decorated with Functional Proteins inside Recombinant Escherichia coli Cells.
abstract: A novel strategy for the preparation of protein-decorated gold nanoparticles (Au NPs) was developed inside Escherichia coli cells, where an artificial oxidoreductase, composed of antibody-binding protein (pG), Bacillus stearothermophilus glycerol dehydrogenase (BsGLD) and a peptide tag with gold-binding affinity (H6C), was overexpressed in the cytoplasm. In situ formation of Au NPs was promoted by a natural electron-donating cofactor, nicotinamide adenine dinucleotide (NAD), which was regenerated to the reduced form of NADH by the catalytic activity of the fusion protein (pG-BsGLD-H6C) overexpressed in the cytoplasm of E. coli, with the concomitant addition of exogenous glycerol to the reaction system. The fusion protein was self-immobilized on Au NPs inside the E. coli cells, which was confirmed by SDS-PAGE and western blotting analyses of the resultant Au NPs. Finally, the IgG binding ability of the pG moiety displayed on Au NPs was evaluated by an enzyme-linked immunosorbent assay.
authors.affiliation: Department of Applied Chemistry, Graduate School of Engineering, Kyushu University.
authors.forename: Yukiho
authors.initials: Y
authors.lastname: Hosomomi
authors.forename: Teppei
authors.initials: T
authors.lastname: Niide
authors.forename: Rie
authors.initials: R
authors.lastname: Wakabayashi
authors.forename: Masahiro
authors.initials: M
authors.lastname: Goto
authors.forename: Noriho
authors.initials: N
authors.lastname: Kamiya
mesh_terms: D055162:Biocatalysis; D003593:Cytoplasm; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D009243:NAD / Q000737:chemistry / Q000235:genetics; D010084:Oxidation-Reduction; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D013401:Sugar Alcohol Dehydrogenases / Q000737:chemistry* / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; D009243:NAD; D006046:Gold; D013401:Sugar Alcohol Dehydrogenases; C020672:glycerol dehydrogenase