Cloning, Expression, and Computational Studying of E. coli L-Asparaginase II to Alter Substrate Specificity

Asparaginase is an enzyme that hydrolyzes the amide bond in L-asparagine and similar amides and possesses expandable anti-cancer properties. In this study, the E. coli L-asparaginase II gene was cloned and the recombinant protein was expressed and its enzymatic activity was measured. To find a mutant with altered substrate specificity, two active site residues were chosen as the starting point for mutagenesis and the effect of the mutation on the enzyme s structure was studied by computational methods

Keywords:

L-asparaginase II , molecular cloning , substrate specificity , protein engineering , molecular modeling