Effect of methanolic extract of some endemic plants from Mazandaran on kinetic parameters of Broccoli’s peroxidase

Broccoli has been well known as a plant of high nutritional value. Application of broccoli peroxidase in different areas of clinical biochemistry, biotechnology and food industry enhances the interest for further study on the enzyme. The objective of this study was to evaluate the kinetic parameters of broccoli's peroxidase and their variations in the presence of methanolic extract of some endemic plants from Mazandaran. In this investigation, the crude extract from broccoli was homogenized, centrifuged, precipitated by ammonium sulfate and dialyzed. The enzyme activity was monitored in 414 nm with H2O2 and ABTS in phosphate-citrate buffer. The specific activity of the peroxidase was determined 0.58 U/mg in crude extract and 3.61 U/mg in dialysate. The optimum pH and temperature of the enzyme were obtained 4 and 30°C respectively. The extract of Artemisia tschernieviana, Eryngium caucasicum, Rosmarinus officinalis, Sambucus ebulus, Ligularia persica, Crataegus douglasii, Papaver rhoeas, Viscum album and Allium ursinum were prepared by maceration method. The enzyme activity was measured in presence of the extracts in different concentrations. The kinetic results shown that the extract of Crataegus douglasii, Rosmarinus officinalis, Papaver rhoeas and Artemisia tschernieviana in 33 µg/ml concentration inhibited the enzyme as 88.2, 79.5, 78.6 and 76.3% respectively. The extract of Ligularia persica shown no effect on maximum velocity of reaction while reduced the Km value by 4 times. Allium ursinu showed a partial inhibition on the peroxidase. Therefore, study on these plants for achieving to effectors on broccoli’s peroxidase, with pharmaceutical, agricultural and industrial applications could be recommended.