RESUMO
Kinetic and physicochemical properties of Moringa oleifera peroxidase purified using a novel and cost efficient protocol was investigated with a view to providing information on its possible biotechnological potentials. Moringa oleifera peroxidase was purified to homogeneity in two steps, involving ATPS and size exclusion chromatography on Sephadex G-100 with a yield of 84.12 %. In-gel activity staining revealed the presence of one isoform of peroxidase. The purified peroxidase is monomeric with native and subunits molecular weight of 38.9 and 43.5 kDa respectively. Kinetic parameters - V max, K m(app) o-dianisidine, K m(app) H 2 O 2 of the purified enzyme were 2.5 units/mg protein, 0.020 ± 0.04 mM and 1.37 ± 0.18 mM respectively. Its optimum pH and temperature were 5 and 30 °C respectively. The purified enzyme cross-linked BSA into an insoluble matrix with the aid of caffeic acid. The study concluded that the purification scheme adopted is rapid and efficient, the purified enzyme exhibited some physiochemical properties that make it suitable for various biotechnological applications.
RESUMO
Datura stramonium seed ranks top among major plants commonly abused as drug in Nigeria. The present study therefore sought to unravel the target organs of toxicity as well as underscore the role of extraction solvent in the toxicity of Datura stramonium seed. Twenty male Wistar rats were randomly placed into four groups (I-IV) of five animals per group. Group I served as the control and was administered with distilled water only, while groups II, III and IV animals received 50â¯mg/kg body weight of aqueous, methanolic and diethylether extracts of Datura stramonium seeds by oral gavage for 14 days. Specific biomarkers of toxicity such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lipid peroxidation (MDA) were estimated in the liver, brain, kidney and heart homogenates as well as serum of experimental animals. Lipid profile and activity of antioxidant enzymes such as superoxide (SOD) and catalase were determined in selected tissues while creatinine and urea were estimated in the kidney and serum. Results indicated that Datura stramonium seed extract generally caused a significant decrease in ALT, AST, ALP and MDA in selected tissue homogenates while these parameters increased significantly in the serum relative to the control group. Lipid profile was significantly deranged across selected tissues while SOD and catalase activity were significantly decreased relative to control. Put together, toxicity of Datura stramonium seed extract is diverse depending on the organ involved and solvent used for its extraction. Therefore, illicit use of Datura stramonium seeds as drugs among young people should be discouraged.
