PURIFICATION AND CHARACTERIZATION OF L-ASPARAGINASE BY PSEUDONOCARDIA ENDOPHYTICA VUK-10 ISOLATED FROM NIZAMPATNAM MANGROVE ECOSYSTEM
Objective: L-asparaginase has been a promising therapeutic agent in the treatment of acute lymphoblastic leukaemia. In the present study a rare actino bacterial strain Pseudonocardia endophytica VUK-10 isolated from Nizampatnam mangrove ecosystem was explored for the production of L-asparaginase.
Methods: The extracellular L-asparaginase enzyme was purified to homogeneity from the P. endophytica VUK-10 strain. The crude culture filtrate was subjected to different purification steps including ammonium sulphate fractionation followed by separation on Sephadex G-100 gel filtration and CM-Sephadex C-50 ion exchange chromatography to obtain a pure enzyme preparation.
Results: The enzyme was purified 96 fold and showed a final specific activity of 702.04 IU/mg with a 61% yield. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed its nature as single peptide chain with molecular weight of 120 kDa. This is the first report on production and purification of L-asparaginase from P. endophytica of mangrove origin.
Conclusion: The extracellular L-asparaginase of the P. endophytica may be effectively used as potential chemotherapeutic agent.
Keywords: Mangrove ecosystem, Pseudonocardia endophytica, L-asparaginase, Purification, SDS-PAGE
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