• Rajesh Kumar Munaganti Acharya Nagarjuna University
  • Vijayalakshmi Muvva Department of Botany and Microbiology, Acharya Nagarjuna University, Guntur 522510, A. P., India
  • Mani Deepa Indupalli Acharya Nagarjuna University


Objectives: To optimize the cultural parameters for improved production of L-asparaginase by Arthrobacter kerguelensis VL-RK_09 isolated from Mango orchards of Vissannapet, Krishna District, A. P., India.

Methods: The strain A. kerguelensis was screened for L-asparaginase production on modified asparagine dextrose salts agar medium. L-asparaginase assay was performed as described by Peterson and Ciegler (1969) with slight modifications. Cell free broth (0.2 ml) was placed with 0.8 ml of 0.05M Tris-HCl and 1 ml of 0.04M L-asparagine and incubated for 15 min at 37oC and liberated ammonia was determined spectrophotometrically at 500 nm by nesslerization. Attempts were made to optimize cultural parameters affecting the production of L-asparaginase by the strain.

Results: Maximal yield of L-asparaginase was recorded after 3days of incubation in asparagine dextrose salts broth with initial pH 7.0 and temperature 30 °C. Biosynthesis of L-asparaginase by the strain was improved (from initial 4.28 IU to 6.5 IU) when cultured in modified asparagine dextrose salts broth (initial pH 7.0) supplemented with 1.5% yeast extract and 2% xylose maintained at 30 ° C for 72 h. This is the first report on the production and optimization of L-asparaginase by A. kerguelensis.

Conclusion: In the present study, the optimal cultural and nutritional conditions for the production of L-asparaginase by A. kerguelensis VL-RK_09 were recorded. This is the first report on the production and optimization of L-asparaginase by A. kerguelensis and further studies on purification and characterization of the enzyme is in progress.


Keywords: Mango orchards, Arthrobacter kerguelensis, L-asparaginase


Download data is not yet available.


1. Warangkar SC, Khobragade CN. Purification, characterization, and effect of thiol compounds on activity of the Erwinia carotovora L-asparaginase. Enzyme Res 2010;1:1-10.
2. Wriston JC, Yellin T. L-asparaginase: a review. Adv Enzimol 1973;39:185.
3. Capizzi RL, Poole M, Cooper MR, Richards F, Stuart JJ, Jakson DV, et al. Treatment of poor risk acute leukaemia with sequential higdone ARA-C and asparaginase. Blood 1984;63:649-700.
4. Broome JD. Evidence that the L-asparaginase activity of guinea pig serum is responsible for its antilymphoma effects. Nature 1961;191:1114-5.
5. Mashburn LT, Wriston JC. Tumor inhibitory effect of L-asparaginase from Escherichia coli. Arch Biochem Biophys 1964;105:450-2.
6. Roberts J, Prager MD, Bachynsky N. The antitumor activity of E. coli L-asparaginase. Cancer Res 1966;26:2213-7.
7. Schwartz JH, Reeves JY, Broome JD. Two L-asparaginase from E. coli and their action against tumors. Proc Natl Acad Sci 1966;56:1516-9.
8. Boyse AE, Old LJ, Campbell HA, Mashburn LT. Suppression of murine leukaemia of various types: comparative inhibition activities of guinea pig serum L-asparaginase and E. coli. J Exp Med 1967;125:17-31.
9. Derst C, Wehner A, Specht V, Rohm KH. States and functions of tyrosine residues in Escherichia coli asparaginase. Eur J Biochem 1994;224:533-40.
10. Mercado L, Arenas G. Escherichia coli L-asparaginase induced phosphorylation of endogenous polypeptide in human immune cells. Sangre 1999;44:438-42.
11. Maladkar NK, Singh VK, Naik SR. Fermentative production and isolation of L-asparaginase from Erwinia cartovora EC-113. Hind Antibiot Bull 1993;35:77-86.
12. Aghaiypour K, Wlodowes A, Lubkowski J. Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase. Biochem 2001;40:5655-64.
13. Mukherjee J, Majumadar S, Scheper T. Studies on nutritional and oxygen requirements for production of L-asparaginase by Enterobacter aerogenes. Appl Microbial Biotechnol 2000;53:180-4.
14. Kil JO, Kim GN, Park I. Extraction of extracellular L-asparaginase from Candida utilis. Biosci Biotechnol Biochem 1995;59:749-50.
15. Muley RG, Sarker S, Ambedkar S, Nail SR. Influence of alkali treated corn steep liquor containing medium on protein production by Staphylococcus aureus. Folia Microbiol 1998:43:31-4.
16. Prista AA, Kyridio DA. L-asparaginase of Thermus thermophilus: purification, properties and identification of essential amino acids for catalytic activity. Mol Cell Biochem 2001:216:93-101.
17. DeJong PJ. L-asparaginase production by Streptomyces griseus. Appl Microbiol 1972:23:1163-4.
18. Mostafa SA, Salma MS. L-Asparaginase producing Streptomyces from the soil of Kuwait. Zentralbl Bakteriol Naturwiss 1979:134:325-34.
19. Gunasekaran S, McDonald L, Manavathu M. Effect of culture media on growth and L-asparaginase production in Nocardia asteroids. Biomed Lett 1995;52:197-201.
20. Abdel-Fatah MK. Studies on the asparaginolytic enzymes of Streptomycetes. Culture conditions for the production of L-asparaginase enzyme from Streptomyces longsporusflavus (F-15) strain. Egypt J Microbiol 1996:30:247-60.
21. Narayana KJP, Kumar KG, Vijayalakshmi M. L-asparaginase production by Streptomyces albidoflavus. Indian J Microbiol 2008:48:331-6.
22. Selvakumar D. Study of L-asparaginase production by Streptomyces noursei MTCC 10469, isolated from marine sponge Callyspongia diffusa. Iran J Biotech 2011;9:102-8.
23. Kavitha A, Vijayalakshmi M. A Study on L-Asparaginase of Nocardia levis MK-VL 113. Sci World J 2012:1-5. doi:10.1100/2012/160434. [Article in Press]
24. Usha Kiranmayi M, Sudhakar P, Vijayalakshmi M. Production and optimization of L-asparaginase by an actinobacterium isolated from Nizampatnam mangrove ecosystem. J Environ Biol 2014;35:799-805.
25. Priya PM, Radhakrishnan M, Balagurunathan M. Production and optimization of L-asparaginase from Streptomyces sp (TA22) isolated from Western Ghats, India. J Chem Pharm Res 2011;3:618-24.
26. Peterson RE, Ciegler A. L-asparaginase production by various bacteria. Appl Microbiol 1969;18:64-7.
27. Tareke E, Rydberg P, Karlsson P, Eriksson S, Tornqvist M. Analysis of acrylamide, a carcinogen formed in heated foodstuffs. J Agric Food Chem 2002;50:4998-06.
28. Verma N, Kumar K, Kaur G, Anand S. E. coli K-12 asparaginase-based asparagine biosensor for leukemia. Artif Cells Blood Substitutes Immobilization Biotechnol 2007;35:449-56.
29. Khamna S, Yokota A, Lumyong S. L-asparaginase production by actinomycetes isolated from some Thai medicinal plant rhizosphere soils. Int J Integr Biol 2009;6:22-6.
30. Abdel-All SM, Abdel-Fatah MK, Khalil A. Studies on the asparaginolytic activity of the brown-pigmented Streptomycetes. J Drug Res 1998;22:171-94.
31. Dhevendaran K, Anithakumari YK. L-asparaginase activity in growing conditions of Streptomyces spp. associated with Therapon jarbua and Villorita cyprinoids of Veli lake, South India. Fish Technol 2002;39:155.
32. Sahu MK, Sivakumar K, Poorani E, Thangaradjou T, Kanna L. Studies on L-asparaginase enzyme of actinomycetes isolated from estuarine fishes. J Environ Biol 2007;28:465-74.
33. Selvam K, Vishnupriya B. Partial purification and cytotoxic activity of L-asparaginase from Streptomyces acrimycini NGP. Int J Res Pharm Biomed Sci 2013;4:859-69.
34. Deshpande N, Choubey P, Agashe M. Studies on optimization of growth parameters for L-Asparaginase Production by Streptomyces ginsengisoli. Sci World J 2014;1-6. doi:10.1155/2014/895167. [Article in Press]
35. Liu FS, Zajic JE. L-Asparaginase synthesis by Erwinia aroideae. Appl Microbiol 1972;23:667-8.
343 Views | 1129 Downloads
How to Cite
Munaganti, R. K., V. Muvva, and M. D. Indupalli. “STUDIES ON OPTIMIZATION OF L-ASPARAGINASE PRODUCTION BY ARTHROBACTER KERGUELENSIS VL-RK_09 ISOLATED FROM MANGO ORCHARDS”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 7, no. 9, July 2015, pp. 112-5,
Original Article(s)