The fabrication of novel uranyl (UO22+) binding protein based sensors is reported. The new biosensor responds to picomolar levels of aqueous uranyl ions within minutes using Lysinibacillus sphaericus JG-A12 S-layer protein tethered to gold electrodes. In comparison to traditional self assembled monolayer based biosensors the porous bioconjugated layer gave greater stability, longer electrode life span and a denser protein layer. Biosensors responded specifically to UO22+ ions and showed minor interference from Ni2+, Cs+, Cd2+ and Co2+. Chemical modification of JG-A12 protein phosphate and carboxyl groups prevented UO22+ binding, showing that both moieties are involved in the recognition to UO22+.
References
[1]
Klaassen, C.D. Casarett and Doull’s Toxicology, 7th ed ed.; McGraw-Hill: New York, NY, USA, 2008; p. 1275.
[2]
Dart, R.C. Medical Toxicology, 3rd ed ed.; Williams&Wilkins: Philadelphia, PA, USA, 2004.
[3]
Keith-Roach, M.J. The speciation, stability, solubility and biodegradation of organic co-contaminant radionuclide complexes: A review. Sci. Total Environ？2008, 396, 1–11.
[4]
Shervedani, R.K.; Farahbakhsh, A.; Bagherzadeh, M. Functionalization of gold cysteamine self-assembled monolayer with ethylenediaminetetraacetic acid as a novel nanosensor. Anal. Chim. Acta？2007, 587, 254–262.
[5]
Cherian, S.; Gupta, R.K.; Mullin, B.C.; Thundat, T. Detection of heavy metal ions using protein-functionalized microcantilever sensors. Biosens. Bioelectron？2003, 19, 411–416.
[6]
Li, S.; Rosen, B.P.; Borges-Walmsley, M.I.; Walmsley, A.R. Evidence for cooperativity between the four binding sites of dimeric ArsD, an As(III)-responsive transcriptional regulator. J. Biol. Chem？2002, 277, 25992–26002.
[7]
Bontidean, I.; Berggren, C.; Johansson, G.; Csoregi, E.; Mattiasson, B.; Lloyd, J.R.; Jakeman, K.J.; Brown, N.L. Detection of heavy metal ions at femtomolar levels using protein-based biosensors. Anal. Chem？1998, 70, 4162–4169.
[8]
Lucarelli, F.; Marrazza, G.; Mascini, M. Enzyme-based impedimetric detection of PCR products using oligonucleotide-modified screen-printed gold electrodes. Biosens. Bioelectron？2005, 20, 2001–2009.
[9]
Yang, Y.; Khoo, S.B. Fabrication of self-assembled monolayer of 8-mercaptoquinoline on polycrystalline gold electrode and its selective catalysis for the reduction of metal ions and the oxidation of biomolecules. Sens. Actuat. B: Chem？2004, 97, 221–230.
[10]
Nakajima, A.; Sakaguchi, T. Selective accumulation of heavy metals by microorganisms. Appl. Microbiol. Biotechnol？1986, 24, 59–64.
[11]
Bienert, G.P.; Schuessler, M.D.; Jahn, T.P. Metalloids: essential, beneficial or toxic? Major intrinsic proteins sort it out. Trends Biochem. Sci？2008, 33, 20–26.
[12]
Pollmann, K.; Raff, J.; Merroun, M.; Fahmy, K.; Selenska-Pobell, S. Metal binding by bacteria from uranium mining waste piles and its technological applications. Biotechnol. Adv？2006, 24, 58–68.
[13]
Pollmann, K.; Raff, J.; Schnorpfeil, M.; Radeva, G.; Selenska-Pobell, S. Novel surface layer protein genes in Bacillus sphaericus associated with unusual insertion elements. Microbiology？2005, 151, 2961–2973.
[14]
Merroun, M.L.; Raff, J.; Rossberg, A.; Hennig, C.; Reich, T.; Selenska-Pobell, S. Complexation of Uranium by Cells and S-Layer Sheets of Bacillus sphaericus JG-A12. Appl. Environ. Microbiol？2005, 71, 5532–5543.
[15]
Sonja, S.-P.; Petra, P.; Vanya, M.; Ivo, B.; Gert, B.; Heino, N. Selective accumulation of heavy metals by three indigenous Bacillus strains, B. cereus, B. megaterium and B. sphaericus, from drain waters of a uranium waste pile. FEMS Microbiol. Ecol？1999, 29, 59–67.
[16]
Hu, S.; Lu, Q.; Xu, Y. Biosensors based on direct electron transfer of protein. In Electrochemical Sensors, Biosensors and their Biomedical Applications; Academic Press: San Diego, CA, USA, 2008; p. 531.
[17]
Jiang, L.; Yu, Z.; Du, W.; Tang, Z.; Jiang, T.; Zhang, C.; Lu, Z. Development of a fluorescent and colorimetric detection methods-based protein microarray for serodiagnosis of TORCH infections. Biosens. Bioelectron？2008, 24, 376–382.
[18]
Chen, H.; Jiang, C.; Yu, C.; Zhang, S.; Liu, B.; Kong, J. Protein chips and nanomaterials for application in tumor marker immunoassays. Biosens. Bioelectron？2009, 24, 3399–3411.
[19]
Chan, C.P.Y.; Wan, T.S.M.; Watkins, K.L.; Pelsers, M.M.A.L.; Van der Voort, D.; Tang, F.P.W.; Lam, K.H.K.; Mill, J.; Yuan, Y.; Lehmann, M.; Hempel, A.; Sanderson, J.E.; Glatz, J.F.C.; Renneberg, R. Rapid analysis of fatty acid-binding proteins with immunosensors and immunotests for early monitoring of tissue injury. Biosens. Bioelectron？2005, 20, 2566–2580.
[20]
Gupta, R.K.; Dobritsa, S.V.; Stiles, C.A.; Essington, M.E.; Liu, Z.Y.; Chen, C.H.; Serpersu, E.H.; Mullin, B.C. Metallohistins: A new class of plant metal-binding proteins. J. Prot. Chem？2002, 21, 529–536.
[21]
Guimar？es-Soares, L.; Felícia, H.; Jo？o Bebianno, M.; Cássio, F. Metal-binding proteins and peptides in the aquatic fungi Fontanospora fusiramosa and Flagellospora curta exposed to severe metal stress. Sci. Total Envir？2006, 372, 148–156.
Debabov, V.G. Bacterial and archaeal S-layers as a subject of nanobiotechnology. Mol. Biol？2004, 38, 482–493.
[24]
Dupres, V.E.A. In vivo Imaging of S-Layer Nanoarrays on Corynebacterium glutamicum. Langmuir Lett？2009, 25, 9653–9655.
[25]
Nelson, A.; Auffret, N.; Borlakoglu, J. Interaction of hydrophobic organic compounds with mercury adsorbed dioleoylphosphatidylcholine monolayers. BBA-Biomembranes？1990, 1021, 205–216.
[26]
Whitehouse, C.; Gidalevitz, D.; Cahuzac, M.; Koeppe, R.E.; Nelson, A. Interaction of Gramicidin Derivatives with Phospholipid Monolayers. Langmuir？2004, 20, 9291–9298.
[27]
Hays, H.C.W.; Millner, P.A.; Prodromidis, M.I. Development of capacitance based immunosensors on mixed self-assembled monolayers. Sens. Actuat. B: Chem？2006, 114, 1064–1070.
[28]
Billah, M.; Hays, H.C.W.; Millner, P.A. Development of a myoglobin impedimetric immunosensor based on mixed self-assembled monolayer onto gold. Microchim. Acta？2008, 160, 447–454.
[29]
Bard, A.J.; Faulkner, L.R. Electrochemical Methods Fundimentals and Applications, 2nd ed ed.; Wiley: Malden, MA, USA, 2001; p. 833.
[30]
Wang, J. Analytical Electrochemistry, 3rd ed ed.; Wiley-VCH: Berlin, Germany, 2006; p. 250.
[31]
Weygand, M.; Wetzer, B.; Pum, D.; Sleytr, U.B.; Cuvillier, N.; Kjaer, K.; Howes, P.B.; L？sche, M. Bacterial S-Layer Protein Coupling to Lipids: X-Ray Reflectivity and Grazing Incidence Diffraction Studies. Biophys. J？1999, 76, 458–468.
[32]
Bergveld, P. A critical evaluation of direct electrical protein detection method. Biosens. Bioelectron？1991, 6, 55–72.
[33]
Campuzano, S.; Pedrero, M.; Montemayor, C.; Fatás, E.; Pingarrón, J.M. Characterization of alkanethiol-self-assembled monolayers-modified gold electrodes by electrochemical impedance spectroscopy. J. Electroanal. Chem？2006, 586, 112–121.
[34]
Fawcett, W.R.; Kovácová, Z.; Motheo, A.J.; Foss, C.A., Jr. Application of the ac admittance technique to double-layer studies on polycrystalline gold electrodes. J. Electroanal. Chem？1992, 326, 91–103.
[35]
Breyer, B.; Bauer, H.H. Electrochemical cells as electrical circuit elements. J. Electroanal. Chem.？, 12, 411–415.
[36]
Rivera-Gandía, J.; Cabrera, C.R. Self-assembled monolayers of 6-mercapto-1-hexanol and mercapto-n-hexyl-poly(dT)18-fluorescein on polycrystalline gold surfaces: An electrochemical impedance spectroscopy study. J. Electroanal. Chem？2007, 605, 145–150.
