Question

Production, purification and characterization of l-asparaginase from pseudomonas aeruginosa 2488 using fermentation

Answer 1

The main objective of the present work is the discovery of new L-asparaginase different from the reported literature but has similar therapeutic effects. The L-asparaginase enzyme was purified to homogeneity from Pseudomonas aeruginosa 2488 cells that were grown on solid-state fermentation. Different purification steps (including ammonium sulfate fractionation followed by separation on Sephadex G-200 gel filtration) were applied to the crude culture filtrate to obtain a pure enzyme preparation and the characterization was followed. The enzyme was purified and showed a final specific activity of 63.8 IU/mg. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed it was one peptide chain with Mr of 42 kDa. The enzyme showed maximum activity at pH 7 when incubated at 37ºC for 30 min. The amino acid composition of the purified enzyme was also determined. So, it can be extremely used in the chemotherapeutic treatment of leukemia by conjugating with PEG Asparginase which is used in cancer chemotherapy.

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