期刊
JOURNAL OF HAZARDOUS MATERIALS
卷 192, 期 2, 页码 794-800出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jhazmat.2011.05.088
关键词
Phytochelatin synthase activity; Cadmium; Glutathione; High performance liquid chromatography with electrochemical detection; Tobacco BY-2 cell suspension
资金
- CEITEC [CZ.1.05/1.1.00/02.0068]
- REMEDTECH [GACR 522/07/0692, GACR 204/09/H002 MSMT 6215712402]
- IGA MENDELU [2/2011]
The synthesis of phytochelatins is catalyzed by gamma-Glu-Cys dipeptidyl transpeptidase called phytochelatin synthase (PCS). Aim of this study was to suggest a new tool for determination of phytochelatin synthase activity in the tobacco BY-2 cells treated with different concentrations of the Cd(II). After the optimization steps, an experiment on BY-2 cells exposed to different concentrations of Cd(NO(3))(2) for 3 days was performed. At the end of the experiment, cells were harvested and homogenized. Reduced glutathione and cadmium (II) ions were added to the cell suspension supernatant. These mixtures were incubated at 35 degrees C for 30 min and analysed using high performance liquid chromatography coupled with electrochemical detector (HPLC-ED). The results revealed that PCS activity rises markedly with increasing concentration of cadmium (II) ions. The lowest concentration of the toxic metal ions caused almost three fold increase in PCS activity as compared to control samples. The activity of PCS (270 fkat) in treated cells was more than seven times higher in comparison to control ones. K(m) for PCS was estimated as 2.3 mM. (C) 2011 Elsevier B.V. All rights reserved.
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