Table 2.
Kinetic parameters of the wild-type CMMase and variants for CMM
The enzymatic reaction was performed in 50 mm sodium acetate buffer (pH 6.0) at 25 °C, and reducing power was measured.
| Enzyme | Km | kcat | kcat/Km |
|---|---|---|---|
| mm | s−1 | s−1 mm−1 | |
| WT | 3.6 ± 0.2 | 59.7 ± 2.9 | 17.8 ± 1.4 |
| Y168A | 45.5 ± 5.3 | 111.1 ± 8.9 | 2.4 ± 0.3 |
| Y204A | 5.0 ± 0.3 | 101.4 ± 2.7 | 20.4 ± 1.5 |
| P203A/Y204N/F205E | 40.8 ± 6.3 | 126.1 ± 12.9 | 3.1 ± 0.6 |
| Y168Q/Y204N | 29.5 ± 2.1 | 146.7 ± 7.0 | 5.0 ± 0.4 |
| Y168Q/P203A/Y204N/F205E | 110.6 ± 56.3 | 45.0 ± 18.6 | 0.4 ± 0.3 |
| C163A | 69.7 ± 5.0 | 74.4 ± 4.4 | 1.1 ± 0.1 |
| C163V | 15.2 ± 0.5 | 32.6 ± 0.6 | 2.1 ± 0.1 |
| C163L | 85.9 ± 14.8 | 97.7 ± 13.3 | 1.1 ± 0.2 |
| C163S | 43.5 ± 5.3 | 33.4 ± 3.0 | 0.8 ± 0.2 |
| S164A | 16.8 ± 0.9 | 127.1 ± 3.9 | 7.9 ± 0.5 |