SINGLE-STEP PURIFICATION OF UREASE FROM CANAVALIA GLADIATA SEEDS
AbstractThe urease (E.C.3.5.1.5) catalyzes the hydrolysis of urea to ammonia and is commonly used in clinical laboratories for the estimation of urea in biological fluids. The urease enzyme of sword bean (Canavalia gladiata) has been purified in single-step by affinity chromatography using epoxy activated agarose containing urea as the affinity ligand. The yield of the purified enzyme was about 83% with specific activity of about 495 U/mg of protein with 14-fold purification. The final preparation had a transparent appearance with free ammonia content less than 0.01 µg/unit and was stable for 15 months at 4-8°C. The enzyme acted optimally at pH 7.5 and 25°C. Thermal stability studies indicated that at pH 7.5 no loss of enzyme activities were recorded up to 35°C for 30 mins. SDS−polyacrylamide gel electrophoretic analysis showed that the purified enzyme was apparently homogeneous and had a molecular weight of approximately 87 kDa. Single band was observed in both native and SDS-PAGE
Article Information
48
845-849
368
2257
English
Ijpsr
ASM Saem, Md. Tanvir Hossain , DHN Chandan , M. Abdul Mottaleb and M. Taufiq Alam
Assistant Professor, Department of Applied Chemistry and Chemical Engineering, Noakhali Science & Technology University, Noakhali-3814, Bangladesh.
tanvir.acce.nstu@gmail.com
05 July, 2014
10 November, 2014
19 January, 2015
http://dx.doi.org/10.13040/IJPSR.0975-8232.6(2).845-49
01 February, 2015
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