Studies on the subsite structure of amylases. III. Inhibition by gluconolactone of the hydrolysis of maltodextrin catalyzed by glucoamylase from Rhizopus niveus.

Inhibition by gluconic acid-1 : 5-lactone (gluconolactone) and phenyl alpha-glucoside of the hydrolysis of maltodextrin catalyzed by glucoamylase [EC 3.2.1.3] from Rhizopus niveus was investigated in relation to the subsite structure of the enzyme. Inhibition by gluconolactone was of the mixed type, whereas that by phenyl alpha-glucoside was purely competitive. These inhibition types were consistent with a theoretical prediction based on the assumption that gluconolactone and phenyl alpha-glucoside bind mainly to Subsites 1 and 2, respectively. The inhibitor constant of gluconolatone was determined to be 1.5 mM, which is in good agreement with the dissociation constant estimated by difference spectrophotometry (1.5 mM) (Ohnishi, M, et al. (1975) J. Biochem, 77, 695-703).