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

Gheorghita Menghiu, Amalia Nicoleta Iancu, Vasile Ostafe, a West University of Timisoara, Faculty of Chemistry, Biology, Geography, Department of Biology - Chemistry, 16 Pestalozzi, 300115 Timisoara, Romania b West University of Timisoara, Multidisciplinary Research Platform "Nicholas Georgescu - Roengen", Advanced Environmental Research Laboratory,4 Oituz, 300086, Timisoara, Romania", "a West University of Timisoara, Faculty of Chemistry, Biology, Geography, Department of Biology - Chemistry, 16 Pestalozzi, 300115 Timisoara, Romania b West University of Timisoara, Multidisciplinary Research Platform "Nicholas Georgescu - Roengen", Advanced Environmental Research Laboratory,4 Oituz, 300086, Timisoara, Romania", "a West University of Timisoara, Faculty of Chemistry, Biology, Geography, Department of Biology - Chemistry, 16 Pestalozzi, 300115 Timisoara, Romania b West University of Timisoara, Multidisciplinary Research Platform "Nicholas Georgescu - Roengen", Advanced Environmental Research Laboratory,4 Oituz, 300086, Timisoara, Romania"
2019 Studia Universitatis Babe?-Bolyai Chemia  
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
more » ... ive bacterium: the peroxidase activity was detected at 4 h after induction of expression.
doi:10.24193/subbchem.2019.2.46 fatcat:x4x2oym3bfhcth2kie362jh6um