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Isolation and partial purification of the clotamine synthetase from the black bean plant (Nigella sativa L.) | ||
| Journal of Education and Science | ||
| Article 18, Volume 22, Issue 1, 2009, Pages 84-95 PDF (0 K) | ||
| DOI: 10.33899/edusj.2009.57421 | ||
| Authors | ||
| Nehal Ezzat* ; Sajeda A. Abood | ||
| Department of Biology, College of Science, University of Mosul, Mosul, Iraq | ||
| Abstract | ||
| ABSTRACT Callus induction occurred on stem segments of Nigella sativa L. seedlings grown on a modified Murashige and Skoog medium containing 10-6 M of 2, 4- dichlorophenoxy acetic acid (2, 4 – D). Glutamine synthetase (EC 6. 3 . 1. 2) was isolated and partially purified from Nigella sativa L. callus the partially purified enzyme from the aqueous extract by ammonium sulfate precipitation, dialysis and gel filtration on sephadex G – 200 revealed that the activity increased by 37 fold. The molecular weight by gel – filtration chromatography was found to be around 50000 Da. The optimum conditions of the partially purified glutamine synthetase from callus were obtained by using Tris – HCl (80 mM) at pH 2.7 as a buffer and 10 mM of glutamate as substrate with incubation temperature 30° C. The Michaelis Menten constants for glutamate, adenosinetriphophate and ammonium chloride were 10 × 10-3M, 2 × 10-3 M and 0.280 × 10-3 M respectively. | ||
| Keywords | ||
| Isolation; clotamine synthetase; black bean plant; Nigella sativa L | ||
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