A fast and sensitive zymography method of peroxidase activity determination using sodium acetate buffer

Abstract

By changing the buffer composition from phosphate to acetate (in both cases, the pH value was 6.0), the sensitivity of the estimation of peroxidase activity in polyacrylamide gels was improved from 4 µg/well to 0.4 µg/well. The optimized method, including electrophoretic separation and zymogram technique realized with guaiacol as substrate, can be performed in only 135 minutes. The improved method was applied to monitor the expression of recombinant peroxidase in a Gram-negative bacterium: the peroxidase activity was detected at 4 h after induction of expression.