Isolation and purification of glutathione peroxidase from hemophilia patients serum and study effect of some Carthamus tinctorius L. (carthamus) secondary metabolites on enzyme activity

Authors

  • Qussay Alhakeem Tikrit University
  • Mohammed Bahry University of Mosul
  • Nazar Ahmed University of Tikrit

Keywords:

Hemophilia, Glutathione peroxidase, purification, Natural product

Abstract

The study was performed measuring the level effectiveness of the Glutathione peroxidase in the serum of Hemophilia patients as well as the separation and purification of this enzyme, as it included 48 samples of serum hemophilia patients of both types (40 samples of type A and 8 of type B), in addition to 50 samples of healthy people only male. The results of this study showed a significant decrease at the probability level P≤ 0.05 of the effectiveness of the Glutathione peroxidase in the serum of hemophilia patients of both types. The Glutathione peroxidase was also separated and purified from the serum of those with hemophilia by precipitation of ammonium sulfate and dialysis, and by using the DEAD - Cellulose exchange chromatography technique where a major protein bundle was relied upon which was used to determine the optimal conditions for the partially purified enzyme.  The optimum conditions were determined for the partially purified enzyme from the blood serum and were maximum activity of (GPx) at minute (6), acidic function at pH = 8, enzyme concentration 105 µg/ml, concentration of substrate H2O2 0.6 mmol/L temperature(40oC) and that the maximum speed (Vmax) and the Michaelis constant (Km) were equal to 0.9μmol/min and 0.195 mmol/Liter, respectively, using a line –weaver plot diagram.

     The enzyme Glutathione peroxidase was also separated and purified from the serum of those with hemophilia by precipitation of ammonium sulfate and dialysis, and by using the DEAD - Cellulose exchange chromatography technique where a major protein bundle was relied upon which was used to determine the optimal conditions for the partially purified enzyme.

      The optimum conditions were determined for the partially purified enzyme from the blood serum and were Maximum activity of (GPx) at minute (6), acidic function at (pH = 8), enzyme concentration (105 µg/ ml), concentration of base material H2O2 (0.6 mmol/L) temperature (40oC) and that the maximum speed (Vmax) and the Michaelis constant (Km) were equal to (0.9μmol / min) and (0.195 mmol / Liter), respectively, using a line –weaver plot diagram.

     The study also included the isolation of the natural product (Oil, Flavonoids, Glycosides) from a seeds of plant Carthamus and the results showed that it increases the enzyme activity  of the partially purified .

 

Published

2020-10-02