TABLE 3.
Activity of mutants of BoAgu115A
| BoAgu115A derivative | Catalytic activitya |
|---|---|
| kcat/Km min−1 m>m−1 | |
| Wild type | 780.0 |
| E162A | ∼780.0b |
| D192A | ∼780.0b |
| W169A | ∼780.0b |
| D478A | ∼780.0b |
| E785A | 198.0 |
| Y788A | 152.0 |
| E782A | 123.0 |
| N205A | 44.0 |
| Y425A | 10.8 |
| K374A | 8.0 |
| Y792A | 7.6 |
| D396N | 5.4 |
| N398A | 3.7 |
| D206A | 2.5 |
| N462A | 1.8 |
| Y373A | 1.1 |
| Y420A | 0.8 |
| H275A | 0.3 |
| W249A | 0.28 |
| E375A | 0.17 |
| H422A | 0.01 |
| R328A | 0.01 |
| H275A/H422A | NDc |
| D332A | ND |
| ΔC-terminal (1–526) | ND |
| ΔC-terminal (1–639) | ND |
| ΔC-terminal (1–665) | ND |
a Wild type and mutants of BoAgu115A were assayed using 1 mm XUXX as the substrate in 20 mm sodium phosphate buffer, pH 7.0, containing 1 mg ml−1 of BSA. At this substrate concentration the initial rate provides a direct readout of the catalytic efficiency of the enzyme.
b ∼780; the activity of the mutants of residues from the other cleft/pocket-like structure on the enzyme was estimated from a single time point reaction.
c ND, no activity could be determined using an assay that can detect activity that is ∼105-fold less than the wild type enzyme.