Detection of heavy metal ions at femtomolar levels using protein-based biosensors.
Sensors based on proteins (GST-SmtA and MerR) with distinct binding sites for heavy metal ions were developed and characterized. A capacitive signal transducer was used to measure the conformational change following binding. The proteins were overexpressed in Escherichia coli, purified, and immobilized in different ways to a self-assembled thiol layer on a gold electrode placed as the working electrode in a potentiostatic arrangement in a flow analysis system. The selectivity and the sensitivity of the two protein-based biosensors were measured and compared for copper, cadmium, mercury, and zinc ions. The GST-SmtA electrodes displayed a broader selectivity (sensing all four heavy metal ions) compared with the MerR-based ones, which showed an accentuated selectivity for mercury ions. Metal ions could be detected with both electrode types down to femtomolar concentration. The upper measuring limits, presumably due to near saturation of the proteins' binding sites, were around 10(-10) M. Control electrodes similarly constructed but based on bovine serum albumin or urease did not yield any signals. The electrodes could be regenerated with EDTA and used for more than 2 weeks with about 40% reduction in sensitivity.
Bontidean
I
I
Department of Biotechnology, Chemical Center, Lund University, Sweden.
Berggren
C
C
Johansson
G
G
Csöregi
E
E
Mattiasson
B
B
Lloyd
J R
JR
Jakeman
K J
KJ
Brown
N L
NL
D000595:Amino Acid Sequence; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D004867:Equipment Design; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019216:Metals, Heavy / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D011199:Potentiometry / Q000379:methods; D014176:Protein Biosynthesis
D019216:Metals, Heavy