Development of a cell surface display system in a magnetotactic bacterium, "Magnetospirillum magneticum" AMB-1.
Bacterial cell surface display is a widely used technology for bioadsorption and for the development of a variety of screening systems. Magnetotactic bacteria are unique species of bacteria due to the presence of magnetic nanoparticles within them. These intracellular, nanosized (50 to 100 nm) magnetic nanoparticles enable the cells to migrate and be manipulated by magnetic force. In this work, using this unique characteristic and based on whole-genomic and comprehensive proteomic analyses of these bacteria, a cell surface display system has been developed by expressing hexahistidine residues within the outer coiled loop of the membrane-specific protein (Msp1) of the "Magnetospirillum magneticum" (proposed name) AMB-1 bacterium. The optimal display site of the hexahistidine residues was successfully identified via secondary structure prediction, immunofluorescence microscopy, and heavy metal binding assay. The established AMB-1 transformant showed high immunofluorescence response, high Cd(2+) binding, and high recovery efficiency in comparison to those of the negative control when manipulated by magnetic force.
Tanaka
Masayoshi
M
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Nakata
Yuko
Y
Mori
Tetsushi
T
Okamura
Yoshiko
Y
Miyasaka
Hitoshi
H
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D000327:Adsorption; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002104:Cadmium / Q000378:metabolism; D002458:Cell Fractionation; D006639:Histidine / Q000235:genetics / Q000378:metabolism; D008280:Magnetics; D042201:Magnetospirillum / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D017433:Protein Structure, Secondary; D014871:Water Microbiology
D001426:Bacterial Proteins; D002104:Cadmium; D006639:Histidine