STUDIES ON OPTIMIZATION OF L-ASPARAGINASE PRODUCTION BY ARTHROBACTER KERGUELENSIS VL-RK_09 ISOLATED FROM MANGO ORCHARDS
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.
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