Title of article :
Expression of recombinant Escherichia coli L-asparaginase II, purification and characterization
Author/Authors :
Maharem, Tahany M. Ain Shams University - Faculty of Science - Department of Biochemistry, Egypt , Sabry, Gilane M. Ain Shams University - Faculty of Science - Department of Biochemistry, Egypt , Mohamed, Mohamed R. Ain Shams University - Faculty of Science - Department of Biochemistry, Egypt , Emam, Manal A. Ain Shams University - Faculty of Science - Department of Biochemistry, Egypt
Abstract :
L-Asparaginase (ASNase) is an anti-cancer (antineoplastic or cytotoxic) chemotherapy drug that is used for the treatment of acute lymphoblastic leukemia (ALL). An efficient and economical scheme was developed for over expression and rapid purification of the Escherichia coli enzyme. The gene encoding for the Escherichia coli L-asparaginase was PCR-amplified and cloned in pGEX-4T1 expression vector. The recombinant L-asparaginase was purified to homogeneity by affinity chromatography on glutathione Sepharose column. The recombinant enzyme had an apparent MW of 152 kDa and a Κm value of 12.5 μM for the main physiological substrate L-asparagine. The pI value was 5.6 while the turnover number (catalytic constant) was 1 x 102 s-1 and the Κcat/Κm value (specificity constant) was 0.8 x 107 M-1s-1
Keywords :
L , asparaginase , II , Escherichia coli , cloning , overexpression affinity purification , acute lymphoblastic leukemia
Journal title :
The Egyptian Journal of Biochemistry and Molecular Biology
Journal title :
The Egyptian Journal of Biochemistry and Molecular Biology
