The rapid purification(less than 1.5 h) of EPSP synthase from bean seedling by Sephadex G 50 and Mono Q chromtography was reported. Specific activity of EPSP synthase obtained by the method was 175.2 nmol·min -1 ...The rapid purification(less than 1.5 h) of EPSP synthase from bean seedling by Sephadex G 50 and Mono Q chromtography was reported. Specific activity of EPSP synthase obtained by the method was 175.2 nmol·min -1 ·mg -1 .Concentrated enzyme solution after adjusting to 50% glycerol( V/V ) and 1mg·mL -1 BSA, was stored at -20℃. EPSP synthase activity was stable at least for 150 days.The activity of EPSP synthase was inhibited approximately 50% by 6.3 μmol·L -1 glyphosate. It showed that the purified EPSP synthase as herbicide screening model is possible. This purified method has been used to study enzyme mechanism of the glyphosate resistant bean.展开更多
文摘The rapid purification(less than 1.5 h) of EPSP synthase from bean seedling by Sephadex G 50 and Mono Q chromtography was reported. Specific activity of EPSP synthase obtained by the method was 175.2 nmol·min -1 ·mg -1 .Concentrated enzyme solution after adjusting to 50% glycerol( V/V ) and 1mg·mL -1 BSA, was stored at -20℃. EPSP synthase activity was stable at least for 150 days.The activity of EPSP synthase was inhibited approximately 50% by 6.3 μmol·L -1 glyphosate. It showed that the purified EPSP synthase as herbicide screening model is possible. This purified method has been used to study enzyme mechanism of the glyphosate resistant bean.
