Characteristics of an amidase isolated from a soil bacterium

Abstract

Amidase was extracted from a bacterium isolated from soil, and its properties were compared with those of amidase in soil. Amidase activity of the bacterial protein was lower than that of soil amidase, respectively, in its optimal pH (7.0 vs 8.5), optimal temperature (50 vs 60°C), K m constant calculated by the Lineweaver-Burk plot (5.6 vs 17.9 mm), activation energy (18.9 vs 43.3 kJmol −) and Q 10 (av. = 1.28 vs 1.75). Bacterial amidase was stable at temperatures ranging from 10 to 50°C and denatured at 55°C. Toluene inhibited both bacterial and soil amidase. When the inhibitions by 21 trace elements were compared by using 2 μmol 0.1 mg −1 protein, the most effective inhibitors of bacterial amidase (> 25% inhibition) were: Ag(I), Cd(II), Cu(II), Hg(II), Ni(II), Pb(II), Zn(II), Al(III) and Se(IV). The effect of 16 pesticides on bacterial amidase varied considerably. By using 2 μg of active ingredient of pesticide 0.1 mg −1 protein, the inhibition of bacterial amidase ranged from 7 to 49% with Dinitroamine and Butylate, respectively. The results show that soil constituents have a considerable influence on the reaction catalyzed by this enzyme.