Multifunctional inorganic-binding beads self-assembled inside engineered bacteria.
Multifunctional shell-core nano/microbeads with a hydrophobic biopolymer core and a designed protein coat for selective binding of an inorganic substance and antibodies were self-assembled inside engineered bacteria. Hybrid genes were constructed to produce tailormade bead-coating proteins in the bacterium Escherichia coli. These fusion proteins contained a binding peptide for an inorganic material, the antibody binding ZZ domain, and a self-assembly promoting as well as biopolymer synthesizing enzyme. Production of these multidomain fusion proteins inside E. coli resulted in self-assembly of beads comprising a biopolyester core and displaying covalently bound binding sites for specific and selective binding of an inorganic substance and any antibody belonging to the immunoglobulin G class. Engineered beads were isolated and purified from the respective E. coli cells by standard cell disruption procedures. Bead morphology and the binding functionalities displayed at the bead surface were assessed by the enzyme-linked immunosorbent assay, transmission electron microscopy, elemental analysis, backscattering electron density, analytical density ultracentrifugation, and atomic force microscopy. These analyses showed that bacteria can be engineered to produce fusion proteins mediating self-assembly of spherical biopolymer beads with binding affinity to gold and/or silica and antibodies. Spherical structures of this type could conceivably serve as nano/microdevices for bioimaging in medical approaches where an antibody mediated targeted delivery of an inorganic contrast agent would be desired.
Jahns
Anika C
AC
Institute of Molecular Biosciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
Haverkamp
Richard G
RG
Rehm
Bernd H A
BH
D000217:Acyltransferases / Q000235:genetics; D000906:Antibodies / Q000378:metabolism; D003287:Contrast Media / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D050939:Gene Fusion; D005818:Genetic Engineering; D006046:Gold / Q000378:metabolism; D007287:Inorganic Chemicals / Q000378:metabolism*; D008863:Microspheres; D053758:Nanoparticles; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012822:Silicon Dioxide / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
D000906:Antibodies; D003287:Contrast Media; D003432:Cross-Linking Reagents; D007287:Inorganic Chemicals; D013205:Staphylococcal Protein A; D006046:Gold; D012822:Silicon Dioxide; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase