Skip to main content

Advertisement

SpringerLink
Log in

Evaluation of Antitumor Activity of Glutaminase-Free Periplasmic Asparaginase from Indigenous Bacterial Isolates as Candidates for Cancer Therapy

  • Research Article
  • Published:
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Asparaginase is a very important antineoplastic drug extensively used for the treatment of acute lymphoblastic leukemia and other tumor malignancies. But intrinsic glutaminase activity of this enzymatic drug is responsible for serious life threatening side effects. In this report, 154 bacterial isolates were isolated from rhizosphere of different plants and river water. All isolates were screened for glutaminase-free periplasmic asparaginase activity, and it was found that only four isolates (i.e., BO1, CO1, CO3 and GG1) lacked detectable glutaminase activity. Their measured asparaginase activities ranged from 0.1 to 0.37 IU/ml. These glutaminase-free asparaginase producing bacterial isolates were identified as Pseudomonas otitidis, Enterobacter cloacae, Ochrobactrum anthropi and Escherichia fergusonii, on the basis of morphological, cultural, biochemical characteristics and 16S rRNA gene sequencing. Crude enzyme extracted from these strains was screened for antitumor activity. Antitumor results showed that asparaginase obtained from P. otitidis and E. cloacae possesses potent antiproliferative effect on human leukemia (MOLT-4) and breast cancer (T47D and MDA-MB-231) cell lines, as compared to standard E. coli asparaginase preparation (Sigma), used in the present study. Based on these results, the asparaginase obtained from P. otitidis and E. cloacae could be used as a potent antiproliferative agent. However, more indepth studies are required for strengthening the current findings.

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

Similar content being viewed by others

References

  1. Kang TS, Stevens RC (2009) Structural aspects of therapeutic enzymes to treat metabolic disorders. Hum Mutat 30:1591–1610

    Article  CAS  PubMed  Google Scholar 

  2. Verma N, Kumar K, Kaur G, Anand S (2007) L-Asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 27:45–62

    Article  CAS  PubMed  Google Scholar 

  3. Asselin BL, Ryan D, Frantz CN et al (1989) In vitro and in vivo killing of acute lymphoblastic leukemia cells by l-asparaginase. Cancer Res 49:4363–4368

    CAS  PubMed  Google Scholar 

  4. Lubkowski J, Palm GJ, Gilliland GL, Derst C, Rohm KH (1996) Crystal structure and amino acid sequence of Wolinella succinogenes l-asparaginase. Eur J Biochem 241:201–207

    Article  CAS  PubMed  Google Scholar 

  5. Gong SS, Basilico C (1990) A mammalian temperature-sensitive mutation affecting G1 progression results from a single amino acid substitution in asparagine synthetase. Nucleic Acids Res 18:3509–3513

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Narta UK, Kanwar SS, Azmi W (2007) Pharmacological and clinical evaluation of L-asparaginase in the treatment of leukemia. Crit Rev Oncol Hematol 61:208–221

    Article  PubMed  Google Scholar 

  7. Geckil H, Gencer S, Ates B, Ozer U, Uckun M, Yilmaz I (2006) Effect of Vitreoscilla hemoglobin on production of a chemotherapeutic enzyme, l-asparaginase, by Pseudomonas aeruginosa. Biotechnol J 2:203–208

    Article  Google Scholar 

  8. Kravchenko OV, Kislitsin YA, Popov AN, Nikonov SV, Kuranova IP (2008) Three dimensional structures of l-asparaginase from Erwinia carotovora complexed with aspartate and glutamate. Acta Crystallogr D 64:248–256

    Article  CAS  PubMed  Google Scholar 

  9. Kumar S, Dasu VV, Pakshirajan K (2011) Purification and characterization of glutaminase-free l-asparaginase from Pectobacterium carotovorum MTCC 1428. Bioresour Technol 102:2077–2082

    Article  CAS  PubMed  Google Scholar 

  10. Geckil H, Ates B, Gencer S, Uckun M, Yilmaz I (2005) Membrane permeabilization of gram-negative bacteria with a potassium phosphate/hexane aqueous phase system for the release of l-asparaginase: an enzyme used in cancer therapy. Process Biochem 40:573–579

    Article  CAS  Google Scholar 

  11. Wriston JC Jr (1070) Asparaginase. Methods Enzymol XVII:732–742

    Google Scholar 

  12. Imada A, Igarasi S, Nakahama K, Isono M (1973) Asparaginase and glutaminase activities of microorganisms. J Gen Microbiol 76:85–99

    Article  CAS  PubMed  Google Scholar 

  13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  14. Mac-Faddin FJ (1980) Biochemical tests for identification of medical bacteria. Williams and Wilkins, Baltimore

    Google Scholar 

  15. Kreig RN, Holt G (1984) Bergey’s manual of systematic bacteriology. William and Wilkins, Baltimore

    Google Scholar 

  16. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  19. Pieters R, Hunger SP, Boos J, Rizzari C, Silverman L, Baruchel A, Goekbuget N, Schrappe M, Pui C (2011) l-Asparaginase treatment in acute lymphoblastic leukemia. Cancer 15:238–249

    Article  Google Scholar 

  20. Jakubas PB, Kulis MK, Giebel S et al (2008) Use of l-asparaginase in acute lymphoblastic leukemia: recommendations of the Polish Adult Leukemia Group. Pol Arch Med Wewn 118:664–669

    Google Scholar 

  21. Ohnuma T, Bergel F, Bray RC (1967) Enzymes in cancer: asparaginase from chicken liver. Biochem J 103:238–245

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Broome JD (1965) Antilymphoma activity of l-asparaginase in vivo: clearance rates of enzyme preparation from guinea pig serum and yeast in relation to their effect on tumor growth. J Natl Cancer Inst 35:967–974

    CAS  PubMed  Google Scholar 

  23. Mashbum LT, Wriston JC (1964) Tumor inhibitory effect of l-asparaginase from Escherichia coli. Arch Biochem Biophys 105:450–452

    Article  Google Scholar 

  24. Yano S, Minato R, Thongsanit J, Tachiki T, Wakayama M (2008) Overexpression of type-I L-asparaginase of Bacillus subtilis in Escherichia coli, rapid purification and characterisation of recombinant type-I l-asparaginase. Ann Microbiol 58:711–716

    Article  CAS  Google Scholar 

  25. Kim KW, Kamerud JQ, Livingston DM, Roon RJ (1988) Asparaginase II of Saccharomyces cerevisiae: characterization of the ASP3 gene. J Biol Chem 263:11948–11953

    CAS  PubMed  Google Scholar 

  26. Reddy VVS, Jayaram HN, Sirsi M, Ramakrishnan T (1969) Inhibitory activity of l-asparaginase from Mycobacterium tuberculosis on Yoshida ascites sarcoma in rats. Arch Biochem Biophys 132:262–267

    Article  CAS  PubMed  Google Scholar 

  27. Cedar H, Schwartz JH (1968) Production of l-asparaginase II by Escherichia coli. J Bacteriol 96:2043–2048

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Oza VP, Parmar PP, Kumar S, Subramanian RB (2010) Anticancer properties of highly purified l-asparaginase from Withania Somnifera L. against Acute Lymphoblastic Leukemia. Appl Biochem Biotechnol 160:1833–1840

    Article  CAS  PubMed  Google Scholar 

  29. Davidson L, Brear DR, Wingard P, Hawkins J, Kitto GB (1977) Purification and properties of an l-glutaminase-l-asparaginase from Pseudomonas acidovoranans. J Bacteriol 129:1379–1386

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

Authors are thankful to Head, Department of P.G. Studies and Research in Biological Science, Rani Durgavati University, Jabalpur for providing Laboratory facilities and Dr. Fayaz A. Malik, Scientist, Department of Pharmacology and Cancer Biology, Indian Institute of Integrative Medicine (IIIM-CSIR), Jammu, India, for providing Laboratory facilities for screening of antitumor activity of enzymes. Authors are also thankful to Madhya Pradesh Biotechnology Council, Bhopal, for providing financial support.

Author information

Authors and Affiliations

  1. Bacteriology Laboratory, Department of P. G. Studies and Research in Biological Science, Rani Durgavati University, Jabalpur, Madhya Pradesh, 482001, India

    Anjana Sharma & Islam Husain

Authors
  1. Anjana Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Islam Husain
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anjana Sharma.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sharma, A., Husain, I. Evaluation of Antitumor Activity of Glutaminase-Free Periplasmic Asparaginase from Indigenous Bacterial Isolates as Candidates for Cancer Therapy. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 87, 997–1004 (2017). https://doi.org/10.1007/s40011-015-0681-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40011-015-0681-z

Keywords

Search

Navigation