Synthetic mechanisms of Cd-induced peptides in fusion yeast

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1. Development of high sensitive determination of PC Phytochelatins were determined by such a way that each PC was separated by HPLC with ODS column, derivatized with DTNB and determined by absorption at 412 nm. It enables to determine glutathione and PCs about 15 sin per a sample.2. Change in PC and its related compounds in culturing Concentrations of PC and its related compounds were measured when S. pombe cells were cultured in the absence of Cd. In the early culturing period the level of PC2 is increased with a concomitant decrease in glutathione. These indicate that PC is synthesized via glutathione.3. Purification of PC-synthsizing enzymeA crude extract solution was prepared from S. poebe cells which were cultured in the absence of Cd. The activity was purified by ammonium sulfate fractionation and hydrophobic chromatography. The fraction containing the synthetic enzyme was applied to a hydroxy appatite column. The activity was, however, lost. In addition the enzyme was found to be very unstable so that a straging method which prevent the enzyme from inactivating is now still on investigation.