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Antimicrobial proteins (AMP) from chickpea (Cicer arietinum L.) seeds were isolated and purified using the saturation of (NH4)2So4 by 80% and gel filtration through Sephacryl S-200, the inhibition zone of the separated peak was 24, and 22 mm for growth of Escherichia coli and Salmonella typhimurium respectively. The molecular mass was 28385 Da estimated through Sephacryl S-200. The optimum pH for activity was 5.5. It was stable at 4.5-7.5, while it lost 21.07 and 55.65% from its activity at pH  3 and 8 respectively, the optimum temperature for activity was 35°C and it was stable at 35°C for 60 min, while it lost all inhibitory activity at 65°C for the same time. The use of AMP at 100 mg resulted in an inhibition zone of 37± 2.92, 35±1.68, 32±2.33, and 33±2.09 mm, with a significant difference at (P≤0.05) against E. coli and S. Typhimurium, Staphylococcus aureus, and Bacillus cereus, respectively. The use of AMP to extend the shelf life of beef patties at 100 resulted in a decrease in the total count of bacteria, as it reached 7.4×102±0.18, 4.6×102±0.22 and 2.8×102±0.19 CFU.g-1, while it was 7.4×102±0.23, 8.2×102±0.31, and 9.5×102±0.27 CFU.g-1 in the control sample during a storage period of 0, 3, and 6 d at 4°C. It was observed that there was no significant difference in the control treatment and AMP added on 0 d, while a significant difference was observed at (P≤0.05) for treatments at a storage period of 3 and 6 d at 4°C.


Antimicrobial proteins (AMPs) Beef patties Chickpea seeds Extending shelf life of food Inhibition of microorganisms

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How to Cite
Alsoufi, M. A. ., & Aziz, R. A. . (2023). Isolation and Purification of Chickpea (Cicer arietinum) Seeds Protein, Testing Their Antibacterial Activity, and Using to Extend The Shelf Life of Beef Patties. Basrah Journal of Agricultural Sciences, 36(2), 59–67.


  1. Aljazy, N. A., Al-Mossawi, A. E.-B. H. J., & Al-Rikabi, A. K. (2019). Study of antibacterial activity of some date seed extracts. Basrah Journal of Agricultural Sciences, 32, 247–257.
  3. Al Akeel, R., Mateen, A., Alharbi, K. K., Alyousef, A. A., Al-Mandeel, H. M., & Syed, R. (2018). Purification and MIC analysis of antimicrobial proteins from Cucumis sativus L. seeds. BMC Complementary Medicine and Therapies, 18, 121.
  5. Al-Sahlany, S. T. G. (2017). Production of biodegradable film from soy protein and essential oil of lemon peel and use it as cheese preservative. Basrah Journal of Agricultural Sciences, 30(2), 27-35.
  7. Al-Soufi, M. A. (2015). Extending the storage life of some fruits by using pullulan produced from locally isolate Aureobasidium pullulans. Iraqi Journal of Market Research and Consumer Protection, 7(1), 179-198.
  9. Alsoufi, M. A., & Aziz, R. A. (2017). Extending shelf life of fruits by using some microorganisms biological products. International Journal of Molecular Biology, 2(5), 00032.
  11. Alsoufi, M. A., & Aziz, R. A. (2019). Use of some plants extracts and pullulan for extending shelf life of apples. Journal of College of Basic Education, 104(25), 653-672.
  13. Alsoufi, M. A., & Aziz, R. A. (2021). Extending the shelf life of food using some biological products. Biochemical and Cellular Archives, 21(2), 4641-4645.
  14. Alsoufi, M. A., & Aziz, R. A. (2022). Use of biopreservation technique to increase shelf life for some types of meat. Bioscience Research, 19(2), 1133-1138
  15. Alsoufi, M.A., Aziz, M. A., Al-Bayati, S. I., & Abbas, S. F. (2016). Purification of red wasp Vespa orientalis and yellow wasp Polistes olivaceaus toxin and estimating some of its biological characteristics. Iraqi Journal of Science, 57(2A), 854-843.
  17. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254.
  19. Brink, I., Šipailienė, A., & Leskauskaitė, D. (2019). Antimicrobial properties of chitosan and whey protein films applied on fresh cut turkey pieces. International Journal of Biological Macromolecules, 130, 810-817.
  21. Chen, C., Li, X., Li, J., Xu, Y., Jing, X., Wu, S., Liu, X., & Zhang, X. (2018). Purification and characterization of an antimicrobial protein from Gastrodia elata Blume tubers. Tropical Journal of Pharmaceutical Research, 17(9), 1717-1723.
  23. Epand, R. F., Maloy, W. L., Ramamoorthy, A., & Epand, R. M. (2010). Probing the charge cluster mechanism in amphipathic helical cationic antimicrobial peptides. Biochemistry, 49, 4076-4084.
  25. Galvez, A., Lopez, R. L., Pulido, R. P., & Burges, G. M. J. (2014). Natural Antimicrobials for Food Preservation. Pp, 3-14. In: Galvez, A., Burges, G. M. J., Lopez, R. L., & Pulido, R. P. (Editors). Food Biopreservation. Springer, New York, 118pp.
  27. Gupta, D., Dubey, J., & Kumar, M. (2016). Phytochemical analysis and antimicrobial activity of some medicinal plants against selected common human pathogenic microorganisms. Asian Pacific Journal of Tropical Disease, 6(1), 15-20.
  29. Kumar, S., Kapoor, V., Gill, K., Singh, K., Xess, I., Das, S. N., & Dey, Sh. (2014). Antifungal and antiproliferative protein from Cicer arietinum: A bioactive compound against emerging pathogens. BioMed Research International, Article ID 387203.
  31. Muhammad, S. M., Sabo, I. A., Gumel, A. M., & Alkali, F. I. (2019). Extraction and purification of antimicrobial proteins from Datura stramonium seed. Journal of Advances in Biotechnology, 18, 1073-1077.
  33. Ningappa, M. B., Dhananjaya, B. L., Dinesha, R., Harsha, R., & Srinivas, L. (2010). Potent antibacterial property of APC protein from curry leaves (Murraya koenigii L.). Food Chemistry, 118, 747-750.
  35. Przybylski, R., Firdaous, L., Châtaigné, G., Dhulster, P., & Nedjar, N. (2016). Production of an antimicrobial peptide derived from slaughterhouse byproduct and its potential application on meat as preservative. Food Chemistry, 211, 306-313.
  37. Rai, M., Pandit, R., Gaikwad, S., & Kövics, G. (2016). Antimicrobial peptides as natural bio-preservative to enhance the shelf-life of food. Journal of Food Science and Technology, 53(9), 3381-3394.
  39. Sakthivel, M. & Palani, P. (2016). Isolation, purification and characterization of antimicrobial protein from seedlings of Bauhinia purpurea L. International Journal of Biological Macromolecules, 86, 390-401.
  41. SAS. (2018). Statistical Analysis System, User's Guide. Statistical. Version 9.6th ed. SAS. Inst. Inc. Cary. N.C.
  42. Silva, H. C., Pinto, L. D. S., Teixeira, E. H., Nascimento, K. S., Cavada, B. S. & Silva, A. L. C. (2014). A novel lectin from Bauhinia ungulata L. seeds with fungi static and antiproliferative activities. Process Biochemistry, 49(3), 203-209.
  44. Silva, M. C. C., Santanaa, L. A., Mentelee, R., Ferreiraa, R. S., de Miranda, A., Silva-Luccac, R. A., Sampaioa, M. U., Correia, M. T. S., & Oliva, M. L. V. (2012). Purification, primary structure and potential functions of a novel lectin from Bauhinia forficata seeds. Process Biochemistry, 47, 1049-1059.
  46. Thapliyal, M., Bisht, A., & Singh, A. (2016). Isolation of antibacterial protein/peptide from Ficus glomerata leaf. International Journal of Current Pharmaceutical Research, 8(4), 24-27.
  48. Zheng, S., Liu, Q., Zhang, G., Wang, H., & Ng, T. B. (2010). Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe sinopica. Acta Biochimica Polonica, 57(1), 43-48.