GLUTAMINASE FREE L-ASPARAGINASE PRODUCING ENDOPHYTES FROM MANGROVES
Objective: To screen endophytes isolated from mangroves for their potential to produce glutaminase free L-asparaginase .
Methods: Endophytes were isolated from leaves and stems of three mangrove plants using surface sterilization technique followed by inoculation of the plant parts in nutrient media. Bacterial and fungal endophytes isolated were tested for production of glutaminase free L-asparaginase by inoculating them in MM9 and MCD media respectively, supplemented with asparagine /glutamine. L-asparaginase activity in glutaminase free L-asparaginase producers was measured by Nesselerization method.
Results: Six bacterial endophytes and one fungal endophyte were found to produce glutaminase free L-asparaginase. Highest L-asparaginase activity was shown by bacterial endophytes of the mangrove Sonneratia caseolaris .
Conclusion: Endophytes isolated in the present study hold the potential to produce glutaminase free L-asparaginase and they need to be considered further in the search of L-asparaginase with high therapeutic index.
1. Goodsell DS. The Molecular Perspective: L-Asparaginase. The Oncologist 2005; 10:238â€“39.
2. Prager MD, Bachynsky N. Asparagine synthetase in asparaginase resistant and susceptible mouse lymphomas. Biochem.Biophys.Res.commun.1968; 31:43-7.
3. Duval M, Suciu S , Ferster A, Rialland X, Nelken B, Lutz P et al .Comparison of Escherichia coli-asparaginase with Erwinia asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research and Treatment of Cancer-Childrenâ€™s Leukemia Group phase 3 trial. Blood 2002; 99:2734â€“39.
4. Ramya LN, Doble M, Rekha VPB, Pulicherla KK. L-Asparaginase as Potent Anti-leukemic Agent and Its Significance of Having Reduced Glutaminase Side Activity for Better treatment of Acute Lymphoblastic Leukaemia. Appl Biochem Biotechnol 2012; 167:2144â€“59.
5. Offman MN, Krol M, Patel N, Krishnan S, Liu J, Saha V, et al. Rational engineering of L-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood 2011;117: 1614â€“21.
6. Chan WK, Lorenzi1 PL, Anishkin A, Purwahal P, Rogers DM, Sukharev S et al. The glutaminase activity of L-asparaginase is not required for anticancer activity
against ASNS-negative cells. Blood 2014; DOI 10.1182/blood-2013-10-535112.
7. Aghaiypour K, Wlodawer A, Lubkowski J. Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase. Biochemistry 2001 ; 40: 5655â€“64.
8. Derst C, Henseling J, Rohm KH. Engineering the substrate specificity of Escherichia coli
asparaginase: II. Selective reduction of glutaminase activity by amino acid replacements at position 248.Protein Science 2000; 9: 2009â€“17.
9. Eden OB, Shaw MP ,Lilleyman JS, Richards S. Non-randomised study comparing toxicity of Escherchia coli and Erwinia asparaginase in children with leukaemia. Med.Pediatric Oncol.1990;18:497-502.
10. Asselin BL, Whitin JC, Coppola DJ, Rupp IP, Sallan SE, Cohen HJ. Comparative pharmacokinetic studies of three asparaginase preparations. J.Clin.Oncol.1993;11:1780-86.
11. Kumar S, Pakshirajan K , Dasu V V. Assessment of physical process conditions for
enhanced production of novel glutaminase-free L-Asparaginase from Pectobacterium carotovorum MTCC 1428. Applied Biochemistry and Biotechnology 2010; 163: 327â€“37.
12. Devi S, Azmi W. One step purification of glutaminase free l-asparaginase from Erwinia carotovora with anticancerous activty. International Journal of Lifescience and Pharma Research 2012; 2(3): 36-45.
13. Mahajan RV, Saran S, Kameswaran K, Kumar V, Saxena R K . Efficient production of L-asparaginase from Bacillus licheniformis with low-glutaminase activity: optimization, scale up and acrylamide degradation studies. Bioresource Technology 2012; 125: 11â€“16.
14. Shakambari G, Sumi BM ,Ashokkumar B, Palanivelu P , Varalakshmi P. Industrial effluent as a substrate for glutaminase free l-asparaginase production from Pseudomonas plecoglossicida strain RS1; media optimization, enzyme purification and its characterization
RSC adv. 2015; 5:48729-38.
15. Bacon CW, White JF. Microbial endophytes.2000. New York, Marcel Dekker, Inc.
16. Shukla ST, Habbu PV, Kulkarni VH, Jagadish KS, Pandey AR and Sutariya VN: Endophytic microbes: A novel source for biologically/pharmacologically active secondary metabolites. Asian Journal of Pharmacology and Toxicology 2014; 2(3): 1-16.
17. Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-asparaginase producing micro organisms, Letters in Applied Microbiology 1997; 24 : 23-26.
18.Thangavel A, Krishnamoorthy G, Subramanian M, Maruthamuthu M. Seaweed Endophytic Fungi: A Potential Source for Glutaminase Free L-Asparaginase. Che Sci Rev Lett 2013; 2(5): 348-54.
19. Imada A, Igarasi S, Nakahama K, Isono M . Asparaginase and glutaminase activities of microorganisms. J .Gen. Microbiol 1973; 76(1): 85â€“99.
20.Nagarajan A, Thirunavukkarasu N, Suryanarayanan TS, Gummadi SN. Screening and Isolation of Novel Glutaminase Free L-asparaginase from Fungal Endophytes. Research Journal of Microbiology 2014; 9:163-76.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.