An Effect of Cadmium and Lead Ions on Escherichia coli with the Cloned Gene for Metallothionein (MT-3) Revealed by Electrochemistry

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Authors	ADAM Vojtěch CHUDOBOVÁ Dagmar TMEJOVÁ Kateřina ČÍHALOVÁ Kristýna KŘÍŽKOVÁ Soňa GURÁŇ Roman KOMÍNKOVÁ Markéta ŽŮREK Michal KREMPLOVÁ Monika JIMENEZ JIMENEZ Ana Maria KONEČNÁ Marie HYNEK David PEKAŘÍK Vladimír KIZEK René
Year of publication	2014
Type	Article in Periodical
Magazine / Source	Electrochimica Acta
MU Faculty or unit	Central European Institute of Technology
Citation
Web	http://www.sciencedirect.com/science/article/pii/S0013468614012730
Doi	http://dx.doi.org/10.1016/j.electacta.2014.06.091
Field	Electrochemistry
Keywords	Escherichia coli; Metallothionein; Voltammetry; Cloning; Metal Ions
Description	This study was focused on the application of electrochemical methods for studying of bacterial strains Escherichia coli and Escherichia coli expressing human metallothionein gene (MT-3) before and after the application of cadmium and/or lead ions in four concentrations (25, 50, 75 and 150 mikroM). Bacterial strains Escherichia coli and Escherichia coli expressing human metallothionein gene (MT-3) were used like model organisms for studying of metals influence to metallothionein expression. Metallothionein was isolated using fast protein liquid chromatography and quantified by electrochemical methods. The occurrence of metallothionein in E.coli was confirmed by gel electrophoresis by the presence of the bands at 15 (MT dimer) and 22 kDa (MT trimer). The changes in electrochemical records due to the interactions of metallothioneins (MT-3 and MT-2A) with cadmium and lead ions showed decline of Cat2 signal of MT with the increasing interaction time because of metal ions binding to cysteines. Electrochemical determination also revealed that Cd(II) remains in E. coli cells in the higher amount than Pb (II). Opposite situation was found at E. coli–MT-3 strain. The antimicrobial effect of cadmium ions was determined by IC50 and was statistically calculated as 39.2 and 95.5 mikroM for E. coli without cloned MT-3 and E. coli carrying MT-3 gene, respectively. High provided concentration IC50 in strains after lead ions application (352.5 mikroM for E. coli without cloning and 207.0 mikroM for E. coli carrying cloned MT-3 gene) indicates lower toxicity of lead ions on bacterial strains compared to the cadmium ions.
Related projects:	CEITEC - Central European Institute of Technology