STUDIES ON OPTIMIZATION OF L-ASPARAGINASE PRODUCTION BY ARTHROBACTER KERGUELENSIS VL-RK_09 ISOLATED FROM MANGO ORCHARDS
Keywords:Mango orchards, Arthrobacter kerguelensis, L-asparaginase
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 Â°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 Â° 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.
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.
Wriston JC, Yellin T. L-asparaginase: a review. Adv Enzimol 1973;39:185.
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.
Broome JD. Evidence that the L-asparaginase activity of guinea pig serum is responsible for its antilymphoma effects. Nature 1961;191:1114-5.
Mashburn LT, Wriston JC. Tumor inhibitory effect of L-asparaginase from Escherichia coli. Arch Biochem Biophys 1964;105:450-2.
Roberts J, Prager MD, Bachynsky N. The antitumor activity of E. coli L-asparaginase. Cancer Res 1966;26:2213-7.
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.
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.
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.
Mercado L, Arenas G. Escherichia coli L-asparaginase induced phosphorylation of endogenous polypeptide in human immune cells. Sangre 1999;44:438-42.
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.
Aghaiypour K, Wlodowes A, Lubkowski J. Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase. Biochem 2001;40:5655-64.
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.
Kil JO, Kim GN, Park I. Extraction of extracellular L-asparaginase from Candida utilis. Biosci Biotechnol Biochem 1995;59:749-50.
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.
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.
DeJong PJ. L-asparaginase production by Streptomyces griseus. Appl Microbiol 1972:23:1163-4.
Mostafa SA, Salma MS. L-Asparaginase producing Streptomyces from the soil of Kuwait. Zentralbl Bakteriol Naturwiss 1979:134:325-34.
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.
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.
Narayana KJP, Kumar KG, Vijayalakshmi M. L-asparaginase production by Streptomyces albidoflavus. Indian J Microbiol 2008:48:331-6.
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.
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]
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.
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.
Peterson RE, Ciegler A. L-asparaginase production by various bacteria. Appl Microbiol 1969;18:64-7.
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.
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.
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.
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.
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.
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.
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.
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]
Liu FS, Zajic JE. L-Asparaginase synthesis by Erwinia aroideae. Appl Microbiol 1972;23:667-8.