Amidolytic Activity of Purified UcB5

To define the group to which the purified UcB5 belongs, the influence of various metallic ions and standard reagents upon its amidolytic activity was explored. In a microplate, these were mixed with 1.0 × 10^–4 mg of the chromogenic substrate S7388 and 2.0 × 10^–3 mg of the enzyme in 100 µl of 20 mM Tris–HCl (pH 8.0) buffer and incubated for 3 min at 37 °C. The released pNA was quantified by the spectrophotometric measurement at A₄₀₅.
For metal ions experimentation, they were tested in parallel tests at the concentration of 5 mM. The applied concentrations of protease reagents varied according to the cited literature (see the results). The enzyme activity in the treatment devoid of reagents and cations was considered 100%.