Table 4. Effect of inhibitors, denaturing and reducing agents on the activity of alkaline metalloprotease.
| Compound | Relative activity (%) c |
|---|---|
| Inhibitor a | |
| None | 100 ± 1 |
| PMSF | 95 ± 2 |
| EDTA | 51 ± 3 |
| Pefabloc SC | 89 ± 1 |
| Denaturing and reducing agent b | |
| None | 100 ± 2 |
| SDS | 6 ± 1 |
| Urea | 38 ± 5 |
| DTT | 24 ± 3 |
| β-mercaptoethanol | 44 ± 2 |
a
A reaction mixture containing 250 μL of 2% (w/v) azocasein in 50 mM Tris/HCl (pH 8.0), 75 μL of AprX, and 1 mM of each inhibitor was incubated at 37 °C for 12 h. The remaining activity was then measured, as described in the text.
b
A reaction mixture containing 250 μL of 2% (w/v) azocasein in 50 mM Tris/HCl (pH 8.0), 75 μL of AprX, and 2% (w/v) SDS, 4 M urea, 0.1% (w/v) DTT, or 0.1% (v/v) β-mercaptoethanol in 50 mM Tris/HCl (pH 8.0) was incubated at 37 °C for 12 h. The remaining activity was then measured, as described in the text.
c
Results show the mean value (n = 3) plus or minus the standard deviation.