Purification, Characterization and Anticancer Activity of L-asparaginase Produced by Marine Aspergillus terreus
2018
Sahar W.M. Hassan | Aida M. Farag | Ehab A. Beltagy
L-asparaginase (E.C.3.5.1.1) is an enzyme responsible for hydrolysis of L-asparagine into aspartic acid andammonia, and has its significant applications in the therapeutics and food technology. It was producedby the marine Aspergillus terreus and precipitated by 65% ammonium sulphate, followed by purificationusing gel filtration on Sephadex G-100 and DEAE-cellulose ion exchange chromatography, which yielded11.96 fold purification. The molecular weight of the purified L-asparaginase was approximately 85kDa, determined by a sodium dodecyl sulphate polyacrylamide gel electrophoresis. L-asparaginaseshowed high affinity for L-asparagine with a Km of 31.5 mM and Vmax of 500 U/ml. The optimum pHand temperature of the purified enzyme were 5.8 and 40 oC, respectively. The L-asparaginase enzymewas stable from pH 4 to 5.8 and stable up to 70 oC. The effect of activators and inhibitors was studiedproviding that CdCl2, Pb Cl2, and Hg Cl2 strongly inhibited the enzyme activity, while Na Cl highly enhancedactivity. Anticancer activity of the purified L-asparaginase was detected against HCT-116, Hep-G2 andMCF-7 cell lines with IC50 ranged from 3.79-12.6 µg/ml.
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