Isolation and characterization of phytochelatin synthase in rice seedlings

Rice plants were treated with 50 microM copper sulfate to induce the synthesis of phytochelatins by means of a series of enzymatic reactions, including that of photochelatin synthase. Phytochelatin synthase extracted from 3-week-old rice seedlings was purified through a series of steps including precipitation with ice-chilled acetone, QAE A-50 anion exchange column, Amicon XM-50 ultrafiltration and Polybuffer Exchange (PBE) 94 chromato-focusing. This enzyme had a molecular mass of about 100 kDa with an isoelectric point of 4.0. The temperature and pH optima of this enzyme were 55 degrees C and pH 7.5, respectively. The enzyme was thermal tolerable and unstable under refrigeration at 4 or -20 degrees C. Cadmium was the most effective stimulator, followed by lead, copper, silver, cobalt and other divalent cations. Calcium and magnesium had no effect.