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Volume: 6, Issue: 5, Sep-Oct, 2018
DOI: 10.7324/JABB.2018.60503

Research Article

Submerged fermentation, purification, and characterization of L-asparaginase from Streptomyces sp. isolated from soil

Savitha S. Desai, Basavaraj S. Hungund

  Author Affiliations


Abstract

Actinomycetes are most valuable microbes because they are reported to produce different kinds of novel antibiotics and bioactive compounds. L-asparaginase enzyme is a chemotherapeutic agent used in the treatment of cancer. The present study reports production and characterization of purified L-asparaginase from an actinomycetes isolate. The isolate (A-164) was found to be a potent producer of L-asparaginase with an enzyme activity of 26.67 international unit (IU)/ml. Biochemical characterization of the isolate indicated that it belongs to genus Streptomyces. Laboratory scale enzyme production studies were undertaken using the standard medium. Further, the enzyme was purified by salt precipitation, anion exchange chromatography, and size exclusion chromatography. The purified enzyme exhibited the specific activity of 390 IU/mg with 60-fold purification and yield of 42% with a molecular weight of 34 KDa. The purified enzyme demonstrated the highest activity at pH 6.0 and temperature 35°C. The enzyme retained its activity till temperature of 50°C, indicating that it is a thermostable enzyme. A study on the effect of various additives on enzyme activity was also undertaken. The Km and Vmax values for the purified L-asparaginase were 0.065 mM and 20.80 IU/ml, respectively. The enzyme can further be used for health care and industrial applications.

Keywords:

L-asparaginase, Streptomyces sp., Biochemical characterization, Chromatography, Sodium dodecyl sulfatepolyacrylamide gel electrophoresis, Kinetic studies.



Citation: Desai SS, Hungund BS. Submerged fermentation, purification, and characterization of L-asparaginase from Streptomyces sp. isolated from soil. J App Biol Biotech. 2018;6(05):17-23. http://doi.org/10.7324/JABB.2018.60503.


Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

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