Purification and Preparation of EPSP Synthase

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.