Author
Environmental Research Center, University of Technology, Baghdad, Iraq
Abstract
The objective of the present study was to extract and purified proliferative factor for peripheral blood lymphocytes from Nigella sativa seeds. Dimeric 44, 32 and 26 -kDa glucosamine-specific lectins were extracted and purified from seeds of N. sativa L. by using different ionic strength solvents (deionized distilled water, phosphate buffer slain and 0.1 M HCl) purification procedure included, precipitation in saturated ammonium sulphate, then Sephadex G-100 gel filtration was used for further purification, finally electrophoresis by polyacrylamide gel to estimate the molecular weight in the presence of sodium dodecylsulphate. Single absorbance peak at 280 nm was recorded in each extracted method. The temperature and pH value on mitogenic activity was studied in temperature ranged from 4°C-60°C, the mitogenic activity reduced with gradual increase of temperatures, 50% of activity lost at temperature greater than 40°C, while the optimum pH for mitogenic activity ranged from 5-9, at pH value less than 3, and greater than 12 the activity disappeared. Interestingly, lectin extracted from N. sativa seeds have high activity especially that extracted with 0.1 M HCl and showed stability at temperature from 4°C to 10°C and pH from 5 to 9. The pure lectin is a homodimeric molecule of 26 kDa with subunit molecular mass of 12.9 kDa. The novel lectin exceed the PHA-Sigma in mitogenic activity.
Keywords
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