Volume 16, number 3
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Extraction, Characterization and Partial Purification of L-Asparaginase from the Leaves of Arachis Hypogaea L.

Niranjana. J, Kandasamy Arun Gandhi*, D. Sunmathi and P. Nanthavanan

Research Department of Biotechnology, Dr. N.G.P. Arts and Science College (Autonomous), Coimbatore, Tamilnadu, India

Corresponding Author E-mail: arungandhicbe@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/2783

ABSTRACT: L-asparaginase has been a promising therapeutic agent in the treatment of acute lymphoblastic leukemia and lymphoma. In recent times, due to the side effects of commercially available bacterial L-asparaginase and its unavoidable importance, plants are being explored as the source of L-asparaginase. The enzyme L-asparaginase was partially purified from Arachis hypogaea L. The crude enzyme extract was subjected to different purification steps including ammonium sulphate precipitation, dialysis followed by separation on Sephadex G-100 gel filtration (size exclusion chromatography) to obtain partially pure form of L - asparaginase. The enzyme was partially purified to 118 folds and contained specific activity of 4686.86 U/mg with 9.85% yield. SDS-PAGE electrophoresis of the partially purified enzyme revealed that it was a single protein with molecular weight of 70 kDa. The study on physiochemical properties showed that L - asparaginase from Arachis hypogaea L. was potassium-dependent in nature, where its optimum pH of enzyme activity was found to be 8.0 and temperature as 40°/50°C with reaction time of 15 - 20 minutes. Also it was observed that the L-asparaginase activity increased with the presence of metal ions such as Na+, Mg++, making it an enzyme dependent on metal ions for its reaction. In addition to this, it was revealed that the enzyme was partially inhibited in presence of certain chelators.  The specificity of  L-asparaginase obtained from Arachis hypogaea L. with lack of  urease activity and minimal glutaminase activity along with less cytotoxicity on human blood indicated it as an efficient chemotherapeutic agent that could be investigated further in future studies.

KEYWORDS: Arachis Hyogaea L; Cytotoxicity; Gel Filtration; Groundnut Plant; L-Asparaginase; Sephadex G-100; Size Exclusion Chromatography

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Niranjana J, Gandhi K. A, Sunmathi D, Nanthavanan P. Extraction, Characterization and Partial Purification of L-Asparaginase from the Leaves of Arachis Hypogaea L. Biosci Biotech Res Asia 2019;16(3).
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Niranjana J, Gandhi K. A, Sunmathi D, Nanthavanan P. Extraction, Characterization and Partial Purification of L-Asparaginase from the Leaves of Arachis Hypogaea L. Biosci Biotech Res Asia 2019;16(3). Available from: https://bit.ly/31LlfJC

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