A catalytic role for threonine‐12 of E. coli asparaginase II as established by site‐directed mutagenesis

Abstract
A threonine‐12 to alanine mutant of E. coli asparaginase II (EC 3.5.1.1) has less than 0.01% of the activity of wild‐type enzyme. Both tertiary and quaternary structure of the enzyme are essentially unaffected by the mutation; thus the activity loss seems to be the result of a direct impairment of catalytic function. As aspartate is still bound by the mutant enzyme, Thr‐12 appears not be involved in substrate binding.