摘要：Phytochelatins are sulphur-rich peptides able to detoxify heavy metals. Their synthesis is catalyzed by the enzyme called phytochelatin synthase. In this study, we used high performance liquid chromatography with electrochemical detection (HPLC-ED) for sensitive determination of phytochelatins and phytochelatin synthase activity. Chromatographic conditions were optimized. Phytochelatin synthase activity was determined in homogenate prepared from cell suspension cultures of tobacco (Nicotiana tabacum cv. BY-2). The enzyme was induced by 50 µM Cd(NO3)2. As a substrate, we used 100 mM reduced glutathione (GSH). Enzyme activity was determined as the equivalent of synthesized phytochelatins after an incubation lasting 15 minutes. Phytochaltins were detected in the reaction mixture by HPLC-ED. The results obtained show that the optimized HPLC-ED method is suitable not only for detection of low concentrations of phytochelatins, but also for determination the activity of phytochelatin synthase.
关键词：glutathione; phytochelatins; phytochelatin synthase; high performance liquid chromatography with electrochemical detection (HPLC-ED).
