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Isolation, screening, optimization and production of Anti-tumor L-Asparaginase by fungi from karwar coastal region

Author Affiliations

  • 1Department of Biotechnology, Government Arts and Science College Karwar-581301, Karnataka, India
  • 2Department of Biotechnology, Government Arts and Science College Karwar-581301, Karnataka, India

Res. J. Recent Sci., Volume 6, Issue (3), Pages 1-7, March,2 (2017)

Abstract

L-asparaginase is an extracellular enzyme that converts L-asparagine to L-Aspartic acid and has gained considerable attention in the recent years. In the present study, soil samples were gathered from different areas in and around the Karwar regions, Karnataka, India. Several fungal species were isolated and identified using standard manuals, screened primarily for the production of L-Asparaginase on Czapek’s Dox medium containing L- Asparagine as main source of carbon. A total of 50 fungal cultures were isolated from soil. The fungal isolates obtained were selected by plate assay method for screening of potential L-Asparaginase production on Modified Czapek Dox’s (mCD) medium. Enzyme production was carried out by submerged fermentation process and was performed by using mCD liquid media. Quantitative enzyme assay was performed to determine the rate of hydrolysis of L-Asparagine by measuring the liberated ammonia by nesslerization. From the 50 total isolates 10 fungal cultures showed L-Asparaginase activities. The cultures were identified as Penicillum species, Basidiomycetes species. Aspergillus species, Mucor species. Fusarium species. Among the above species Aspergillus sp showed potential L-Asparaginase production (155U/ml). An attempt is made to optimize the cultural conditions for the production of potential L-Asparaginase by using submerged fermentation. Different temperature (150C, 250C, 350C, 450C) different pH (3.5, 5.5, 7.5, 9.5, and 11.5) different carbon and nitrogen source were used. The highest amount of enzyme production was observed at pH 7.5(155 U/ml) and temperature at 35°C (160 U/ml), among the various carbon sources dextrose promoted maximum enzyme activity (176 U/ml) and highest activity was obtained when nitrogen source ammonium sulphate was used (185 U/ml).

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