Skip to main content
SpringerLink
Log in

Anticancer Properties of Highly Purified l-Asparaginase from Withania somnifera L. against Acute Lymphoblastic Leukemia

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Withania somnifera L. has been traditionally used as a sedative and hypnotic. The present study was carried out for the purification, characterization, and in vitro cytotoxicity of l-asparaginase from W. somnifera L. l-Asparaginase was purified from the fruits of W. somnifera L. up to 95% through chromatography. The purified l-asparaginase was characterized by size exclusion chromatography, polyacrylamide gel electrophoresis (PAGE), and 2D PAGE. The antitumor and growth inhibition effect of the l-asparaginase was assessed using [3-(4, 5-dimethyl-thiazol-2yl)-2, 5-diphenyl-tetrazolium bromide] (MTT) colorimetric dye reduction method. The purified enzyme is a homodimer, with a molecular mass of 72 ± 0.5 kDa, and the pI value of the enzyme was around 5.1. This is the first report of the plant containing l-asparaginase with antitumor activity. Data obtained from the MTT assay showed a LD50 value of 1.45 ± 0.05 IU/ml. W. somnifera L. proved to be an effective and a novel source of l-asparaginase. Furthermore, it shows a lot of similarity with bacterial l-asparaginases EC-2.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Ichikawa, H., Takada, Y., Shishodia, S., Jayaprakasam, B., Nair, M. G., & Aggarwal, B. B. (2006). Molecular Cancer Therapeutics, 5(6), 1434–1445. doi:10.1158/1535-7163.MCT-06-0096.

    Article  CAS  Google Scholar 

  2. Cammack, K., & Marlborough, D. (1972). Biochemistry Journal, 126, 361.

    CAS  Google Scholar 

  3. Borek, D., & Jaskolski, M. (2001). Sequence analysis of enzymes with asparaginase activity. Acta Biochimica Polonica, 48, 893–902.

    CAS  Google Scholar 

  4. Ortlund, E., Lacount, M. W., Lewinski, K., & Lebioda, L. (2000). Biochemistry, 39, 1199–1204. doi:10.1021/bi991797d.

    Article  CAS  Google Scholar 

  5. Bonthron, D. T., & Jaskolski, M. (1997). Acta Biochimica Polonica, 44, 491–504.

    CAS  Google Scholar 

  6. Sugimoto, H., Odani, S., & Yamashita, S. (1998). Journal of Biological Chemistry, 273, 12536–12542. doi:10.1074/jbc.273.20.12536.

    Article  CAS  Google Scholar 

  7. Tumbula, D. L., Becker, H. D., Chang, W. Z., & Soll, D. (2000). Nature, 407, 106–110. doi:10.1038/35024120.

    Article  CAS  Google Scholar 

  8. Lubkowski, J., Dauter, M., Aghaiypour, K., Wlodawer, A., & Dauter, Z. (2003). Acta Crystallographica D, 59, 84–92. doi:10.1107/S0907444902019443.

    Google Scholar 

  9. Lee, B., & Yang, H. (1973). Journal of Biological Chemistry, 248, 7620.

    CAS  Google Scholar 

  10. Jones, P., Kristiansen, T., & Einarsson, M. (1973). Biochimica et Biophysica Acta, 327, 146.

    Google Scholar 

  11. Whelan, H., & Wriston, J. (1974). Biochimica et Biophysica Acta, 365, 212.

    CAS  Google Scholar 

  12. Soru, E., Teodorescu, M., Zaharia, O., Szabados, J., & Rudescu, K. (1972). Canadian Journal of Biochemistry, 50, 1149. doi:10.1139/v72-182.

    Article  CAS  Google Scholar 

  13. DeJong, P. (1972). Applied Microbiology, 23, 1163.

    CAS  Google Scholar 

  14. Roberts, J., Holcenberg, J., & Dolowy, W. (1972). Journal of Biological Chemistry, 247, 84.

    CAS  Google Scholar 

  15. Mathews, W., & Brown, H. (1974). Enzyme, 17, 276.

    Google Scholar 

  16. Siechiechowicz, K., & Ireland, R. (1989). Phytochemistry, 28, 2275. doi:10.1016/S0031-9422(00)97967-6.

    Article  Google Scholar 

  17. Lough, T. J., Reddington, B. D., Grant, M. R., Hill, D. F., Reynolds, P. H. S., & Farnden, K. J. F. (1992). Plant Molecular Biology, 19, 391–399. doi:10.1007/BF00023386.

    Article  CAS  Google Scholar 

  18. Bruneau, L., Chapman, R., & Marsolais, F. (2006). Planta, 224, 668–679. doi:10.1007/s00425-006-0245-9.

    Article  CAS  Google Scholar 

  19. Sodek, L., Lea, P. J., & Miflin, D. J. (1980). Plant Physiology, 65, 22–26. doi:10.1104/pp.65.1.22.

    Article  CAS  Google Scholar 

  20. Sieciechowicz, K. A., Joy, K. W., & Ireland, R. J. (1988). Phytochemistry, 27, 663–671. doi:10.1016/0031-9422(88)84071-8.

    Article  CAS  Google Scholar 

  21. Laemmli, U. (1970). Nature, 227, 680–685. doi:10.1038/227680a0.

    Article  CAS  Google Scholar 

  22. Blum, H., & Gross, H. J. (1987). Electrophoresis, 8, 93–99. doi:10.1002/elps.1150080203.

    Article  CAS  Google Scholar 

  23. Moriji, M., Masami, M., Kumi, K., Akimitsu, M., Tadaaki, U., & Kazumasa, Y. (1970). Cancer Research, 30, 768–772.

    Google Scholar 

  24. Carmichael, J., DeGraff, W. G., & Gazdar, A. F. (1987). Cancer Research, 47, 936–942.

    CAS  Google Scholar 

  25. Mosmann, T. (1983). Journal of Immunological Methods, 65, 55–63. doi:10.1016/0022-1759(83)90303-4.

    Article  CAS  Google Scholar 

  26. Lea, P. J., Festenstein, G. N., Hughes, G. R., & Miflin, B. J. (1984). Phytochemistry, 23, 511–514. doi:10.1016/S0031-9422(00)80369-6.

    Article  CAS  Google Scholar 

  27. Mozeena, B. V., & Sivaramakrisnan. (1980). Journal of Biosciences, 2, 291–297. doi:10.1007/BF02716861.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Biotechnology, BRD School of Biosciences, Sardar Patel University, Sardar Patel Maidan, Vadtal Road, Satellite Campus, Post Box No. 39, Vallabh Vidyanagar, 388 120, Gujarat, India

    Vishal P. Oza, Pritesh P. Parmar, Sushil Kumar & R. B. Subramanian

Authors
  1. Vishal P. Oza
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pritesh P. Parmar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sushil Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. B. Subramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R. B. Subramanian.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Oza, V.P., Parmar, P.P., Kumar, S. et al. Anticancer Properties of Highly Purified l-Asparaginase from Withania somnifera L. against Acute Lymphoblastic Leukemia. Appl Biochem Biotechnol 160, 1833–1840 (2010). https://doi.org/10.1007/s12010-009-8667-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-009-8667-z

Keywords

Search

Navigation