"Employment of HPLC with coulometric detection for analysis of phytochelatin synthase activity"@en . . . . . . "Employment of HPLC with coulometric detection for analysis of phytochelatin synthase activity" . . "Brno" . . . . . "Kry\u0161tofov\u00E1, Olga" . "Employment of HPLC with coulometric detection for analysis of phytochelatin synthase activity"@en . "MENDELU" . "Zehn\u00E1lek, Josef" . "P(GA522/07/0692), P(GD204/09/H002)" . "RIV/62156489:43210/11:00174192" . "Adam, Vojt\u011Bch" . "Employment of HPLC with coulometric detection for analysis of phytochelatin synthase activity" . . . . . "Phytochelatins are physiologically active peptides which have to be synthesized by phytochelatin synthase (PCS). The main aim of this study was to optimize high performance liquid chromatography coupled with electrochemical detector (HPLC-ED) as suitable tool for determination of the phytochelatin synthase activity. Coulometric detector with porous graphite working electrode was used. Firstly we optimized a separation of two compounds which are important for monitoring of PCS activity. It was glutathione reduced as substrate for the reaction and fytochelatin-2 as a product. PCS is the best activated by cadmium ions. We used a model with BY-2 tobacco cells. We conducted the in vivo 3 day cultivation experiment where BY-2 cells were treated by various concentrations of cadmium. We then homogenized the cells and immediately analyzed the extracts by optimized HPLC-ED method. Moreover we observed that with higher concentrations of applied cadmium there was increasing of amount of PC-2. The lowest concentration of the toxic metal ions caused almost three times enhancing PCS activity compared to control samples."@en . . . "XI. pracovn\u00ED setk\u00E1n\u00ED fyzik\u00E1ln\u00EDch chemik\u016F a elektrochemik\u016F" . . "4"^^ . . "Phytochelatins are physiologically active peptides which have to be synthesized by phytochelatin synthase (PCS). The main aim of this study was to optimize high performance liquid chromatography coupled with electrochemical detector (HPLC-ED) as suitable tool for determination of the phytochelatin synthase activity. Coulometric detector with porous graphite working electrode was used. Firstly we optimized a separation of two compounds which are important for monitoring of PCS activity. It was glutathione reduced as substrate for the reaction and fytochelatin-2 as a product. PCS is the best activated by cadmium ions. We used a model with BY-2 tobacco cells. We conducted the in vivo 3 day cultivation experiment where BY-2 cells were treated by various concentrations of cadmium. We then homogenized the cells and immediately analyzed the extracts by optimized HPLC-ED method. Moreover we observed that with higher concentrations of applied cadmium there was increasing of amount of PC-2. The lowest concentration of the toxic metal ions caused almost three times enhancing PCS activity compared to control samples." . "Beklov\u00E1, Miroslava" . "[C7DD46FF9C9A]" . "\u0160obrov\u00E1, Pavl\u00EDna" . . . "43210" . "RIV/62156489:43210/11:00174192!RIV12-GA0-43210___" . . "2011-01-01+01:00"^^ . "7"^^ . "7"^^ . "Z\u00EDtka, Ond\u0159ej" . . "197422" . "phytochelatin synthase; coulometric analysis; HPLC"@en . "978-80-7375-514-0" . "Mendelova zem\u011Bd\u011Blsk\u00E1 a lesnick\u00E1 univerzita v Brn\u011B" . "Kizek, Ren\u00E9" . . .