ACTIVITY OF ASPARAGINASE, PURIFIED AND CHARACTERIZED FROM FROZEN SEEDS OF PISUM SATIVUM.

  • Ministry of Environment/ Republic of Iraq.
  • Biotechnology Research Center, Al - Nahrain University, Baghdad, Iraq.
  • College of Veterinary Medicine, Al-Muthanna University, Samawa City, Al-Muthanna Province, Iraq.
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Asparaginase was extracted from Frozen Plant seeds of Pisam sativum collected from a field crop after 3 months of freezing. Enzyme specific activity was (74.1 U/mg). Asparaginase was extracted from frozen seeds and purified by ion exchange chromatography using DEAE-Cellulose and gel filtration chromatography using sephadex G-200. We found that the molecular weight of asparaginase is 44000 Dalton, specific activity of Purified asparaginase was 226.5 U/mg. Purified asparaginase was characterized, the optimum pH for enzyme activity and stability was pH 8.5 and 9.5 respectively, while the optimum temperature for enzyme activity and stability was 40 ºC and 37 ºC respectively.

  1. Borek, and Jaskolski, M. (2001). Sequence analysis of enzymes with asparaginase activity. Acta Biochem. Polon. J., 48:893-902.
  2. Sieciechowicz, A.; Joy, K. W. and Irleland, R. J. (1988). The Metabolism of Asparaginase in Plants. Phytochem. J., 27(3):633-671.
  3. Faostat (2008). Available at http://www.fao.org Fernandez-Aparicio M., Perez-De-Luque A., Prats E., Rubiales D. (2008a): Variability of interactions between barrel medic (Medicago truncatula) genotypes and Orobanche species. Annals Applied Biology, 153: 117–126.
  4. Davies, D. R., Berry, G. J. Heath, M. C.  and Dawkins. T. C. K.  (1985). Pea (Pisum sativum L.). In: R.J. Summer field and EH Roberts, (eds.), Williams Collins Sons and Co. Ltd, London, UK. 147-198.
  5. Ho,Y., Y.T. Lin, R.G. Labbe and K. Shetty. 2003. Antimicrobial activity of sprouted pea and chickpea extracts against Helicobacter pylori.  http://ift.confex.com/ift/  2003/techprogram/paper_17775.htm.
  6. Amarowicz, R., Karamac, M. and Weider, S. (2001). Antioxidant activity of phenolic fraction of pea (Pisum sativum). Czech. J. Food Sci. 19(4): 139-142.
  7. Al-Hussaini, K. S, 2012. Effect of blanching and packages in content vitamin C in green beans and green peas frozen for different Periods. Al-kofaa Journal for agricultural sciences. 4(1): 23-30.
  8. Chang, S. and Farnden, K. J. (1981). Purification and Properties of asparaginase from Lupinus arboreus and Lupinus angustifolius. Arch. Biochem. Biophys. J., 208:49-58.
  9. Ren, ; He. F. and Zhang, L. (2010). The construction and application of a new PPY-MSPQC for L-asparaginase activity assay. Sensors and Actuators. J., 145: 272-277.
  10. Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. J., 72:248-254.
  11. Segel, H. (1976). Biochemical Calculation. 2th edition. John Wiley and sons. New York.
  12. Andrews, (1970). Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods of Biochem. Anal., 18:1-53.
  13. Khalaf, Z. A., Al-Ani, N. K., and Jasim, H. M. (2012). Optimum conditions for asparaginase extraction from Pisum sativum subspp Jof,. Iran Journal of Plant Physiology, vol. 2,: 517–521.
  14. Khalaf, Z. A., (2012). Production, purification and characterization of asparaginase from Pisum sativum var.Jof. MS.c. thesis, Collage of science, Al-Nahrain University. Iraq.
  15. Sodek, ; Lea, P. J. and Miflin, B. J. (1980). Distribution and Properties of a potassium-dependent asparaginase isolated from developing seeds of Pisum sativum and other plants. Plant Physiol. J., 65: 22-26.
  16. Chagaz, P. and Sodek, L. (2001). Purification and Properties of asparaginase from the testa of immature seeds of pea (Pisum sativum L.) Int. J. Braz. Arch. of Biol. and Technol. 239-244.
  17. Mckee, R. and Mckee, T. (1996). Biochemistry: the molecular Basis of life. (3rd edition). Mcgraw Hill. P:121.
  18. Bezkorovainy, and Rafelson, M. F., (2007). Concise Biochemistry. Marcel Dekker, Inc. New York. Pp:108-111.
  19. Saleh A. Mohamed, Mohamed F. Elshal, Taha A. Kumosani, and AliaM. Aldahlawi, (2015). Purification and Characterization of Asparaginase from Phaseolus vulgaris Seeds. Hindawi Publishing Corporation. Evidence-Based Complementary and Alternative Medicine. Volume 2015, Article ID 309214,:6-13

[Zena Abdullah Khalaf, Ayat Adnan Abbas and Ali Mosa Rashid Al-Yasari. (2017); ACTIVITY OF ASPARAGINASE, PURIFIED AND CHARACTERIZED FROM FROZEN SEEDS OF PISUM SATIVUM. Int. J. of Adv. Res. 5 (Mar). 1188-1194] (ISSN 2320-5407). www.journalijar.com

Zena Abdullah khalaf
Researcher of the ministry of Environment

DOI:

Article DOI: 10.21474/IJAR01/3622      
DOI URL: https://dx.doi.org/10.21474/IJAR01/3622

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