PMID:22286990

Citation	Sedlak, RH, Hnilova, M, Grosh, C, Fong, H, Baneyx, F, Schwartz, D, Sarikaya, M, Tamerler, C and Traxler, B (2012) Engineered Escherichia coli silver-binding periplasmic protein that promotes silver tolerance. Appl. Environ. Microbiol. 78:2289-96
Abstract	Silver toxicity is a problem that microorganisms face in medical and environmental settings. Through exposure to silver compounds, some bacteria have adapted to growth in high concentrations of silver ions. Such adapted microbes may be dangerous as pathogens but, alternatively, could be potentially useful in nanomaterial-manufacturing applications. While naturally adapted isolates typically utilize efflux pumps to achieve metal resistance, we have engineered a silver-tolerant Escherichia coli strain by the use of a simple silver-binding peptide motif. A silver-binding peptide, AgBP2, was identified from a combinatorial display library and fused to the C terminus of the E. coli maltose-binding protein (MBP) to yield a silver-binding protein exhibiting nanomolar affinity for the metal. Growth experiments performed in the presence of silver nitrate showed that cells secreting MBP-AgBP2 into the periplasm exhibited silver tolerance in a batch culture, while those expressing a cytoplasmic version of the fusion protein or MBP alone did not. Transmission electron microscopy analysis of silver-tolerant cells revealed the presence of electron-dense silver nanoparticles. This is the first report of a specifically engineered metal-binding peptide exhibiting a strong in vivo phenotype, pointing toward a novel ability to manipulate bacterial interactions with heavy metals by the use of short and simple peptide motifs. Engineered metal-ion-tolerant microorganisms such as this E. coli strain could potentially be used in applications ranging from remediation to interrogation of biomolecule-metal interactions in vivo.
Links	PubMed PMC3302632 Online version:10.1128/AEM.06823-11
Keywords	Batch Cell Culture Techniques; Biotechnology/methods; Escherichia coli/drug effects; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Genetic Engineering/methods; Maltose-Binding Proteins/genetics; Maltose-Binding Proteins/metabolism; Metals, Heavy/metabolism; Metals, Heavy/pharmacology; Microbial Sensitivity Tests; Peptides/genetics; Peptides/metabolism; Periplasmic Proteins/genetics; Periplasmic Proteins/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Silver/metabolism; Silver/pharmacology; Silver Nitrate/metabolism; Silver Nitrate/pharmacology
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ECOLI:MALE		GO:1990060: maltose transport complex	ECO:0000314:		C		Figure 7	complete CACAO 8760

References

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