Abstract
BackgroundUrease, one of the highly efficient known enzymes, catalyzes the hydrolysis of urea into ammonia and carbon dioxide. The present study aimed to extract urease from pea seeds (Pisum Sativum L). The enzyme was then purified in three consequence steps: acetone precipitation, DEAE-cellulose ion-exchange chromatography, and gel filtration chromatography (Sephacryl S-200 column).ResultsThe purification fold was 12.85 with a yield of 40%. The molecular weight of the isolated urease was estimated by chromatography to be 269,000 Daltons. Maximum urease activity (190 U/g) was achieved at the optimum conditions of 40°C and pH of 7.5 after 5 min of incubation. The kinetic parameters, K m and V max , were estimated by Lineweaver-Burk fits and found to be 500 mM and 333.3 U/g, respectively. The thermodynamic constants of activation, ΔH, E a , and ΔS, were determined using Arrhenius plot and found to be 21.20 kJ/mol, 23.7 kJ/mol, and 1.18 kJ/mol/K, respectively.ConclusionsUrease was purified from germinating Pisum Sativum L. seeds. The purification fold, yield, and molecular weight were determined. The effects of pH, concentration of enzyme, temperature, concentration of substrate, and storage period on urease activity were examined. This may provide an insight on the various aspects of the property of the enzyme. The significance of extracting urease from different sources could play a good role in understanding the metabolism of urea in plants.
Highlights
Urease, one of the highly efficient known enzymes, catalyzes the hydrolysis of urea into ammonia and carbon dioxide
Several reports have been published on the extraction of urease from various bacteria [2,3,4], and plants [5,6,7,8,9,10,11]
Pisum Sativum L. seeds were obtained from faculty of agriculture, Kafr Elshaikh University, Kafrelshaikh city, Egypt
Summary
One of the highly efficient known enzymes, catalyzes the hydrolysis of urea into ammonia and carbon dioxide. Catalyzing the hydrolysis of urea into ammonia and carbon dioxide, ureases (urea amidohydrolases, EC 3.5.1.5) are a one of known highly efficient enzymes that belong to amidohydrolase and phosphotriesterase superfamily [1]. Several reports have been published on the extraction of urease from various bacteria [2,3,4], and plants [5,6,7,8,9,10,11]. The plant and fungal ureases are homo-oligomeric with identical proteins repetition. The crystal structure of protein is often the key to its enzyme function. This configuration is governed by its primary structure and environment. That alters the shape of the enzyme or blocks the access to the active site in substrate, will affect enzyme activity
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