Isolation and partial purification of the clotamine synthetase from the black bean plant (Nigella sativa L.)

• Main Authors: Nehal Ezzat, Sajeda Abood
• Format: Article
• Language: Arabic
• Published: College of Education for Pure Sciences 2009-03-01
• Series: مجلة التربية والعلم
• Subjects: isolation; clotamine synthetase; black bean plant; nigella sativa l
• Online Access: https://edusj.mosuljournals.com/article_57421_81db50f49564b4fbb06a3f68cb81e240.pdf
• ISSN: 1812-125X; 2664-2530

ABSTRACT <br /> 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). <br /> 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.