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.