Table. 2.
Proteins | Kinetic parameters | Substratea | ||
---|---|---|---|---|
Ampicillin | Cefotaxime | Imipenem | ||
NDM-1 | KM (μM) | 143 ± 3 | 6 ± 1 | 65 ±± 10 |
kcat (s−1) | 550 ± 15 | 85 ± 16 | 238 ± 16 | |
kcat/KM (μM−1 s−1) | 3.84 ± 0.18 | 13.79 ± 1.49 | 3.71 ± 0.36 | |
F64H | KM (μM) | 188 ± 31 | 6 ± 0.1 | 164 ± 19 |
kcat (s−1) | 793 ± 24 | 83 ± 2 | 409 ± 15 | |
kcat/KM (μM−1 s−1) | 4.27 ± 0.57 | 14.80 ± 0.62 | 2.50 ± 0.20 | |
D84E | KM (μM) | 82 ± 6 | 16 ± 0.2 | 70 ± 5 |
kcat (s−1) | 597 ± 17 | 113 ± 3 | 153 ± 3 | |
kcat/KM (μM−1 s−1) | 7.29 ± 0.29 | 6.77 ± 0.23 | 2.20 ± 0.12 | |
W87P | KM (μM) | 518 ± 36 | 6 ± 1 | 295 ± 54 |
kcat (s−1) | 1823 ± 8 | 46 ± 3 | 351 ± 49 | |
kcat/KM (μM−1 s−1) | 3.53 ± 0.23 | 8.50 ± 1.84 | 1.20 ± 0.05 | |
K121R | KM (μM) | 61 ± 3 | 4 ± 0.4 | 31 ± 9 |
kcat (s−1) | 26 ± 0.35 | 13 ± 0.3 | 36 ± 2 | |
kcat/KM (μM−1 s−1) | 0.43 ± 0.03 | 3.69 ± 0.31 | 1.20 ± 0.28 | |
K121T | KM (μM) | 231 ± 0.71 | 10 ± 2 | 117 ± 13 |
kcat (s−1) | 246 ± 8 | 50 ± 0.3 | 50 ± 2 | |
kcat/KM (μM−1 s−1) | 1.06 ± 0.03 | 5.13 ± 0.79 | 0.43 ± 0.03 | |
G195A | KM (μM) | 57 ± 14 | 4 ± 0.5 | 52 ± 15 |
kcat (s−1) | 248 ± 24 | 32 ± 0.04 | 94 ± 11 | |
kcat/KM (μM−1 s−1) | 4.43 ± 0.67 | 7.74 ± 0.85 | 1.84 ± 0.31 | |
D199E | KM (μM) | 11 ± 2 | 6 ± 0.4 | 6 ± 0.8 |
kcat (s−1) | 23 ± 2 | 48 ± 0.7 | 17 ± 1 | |
kcat/KM (μM−1 s−1) | 2.10 ± 0.12 | 7.65 ± 0.40 | 2.81 ± 0.18 | |
K224H | KM (μM) | 906 ± 26 | 26 ± 0.6 | ND |
kcat (s−1) | 330 ± 6 | 111 ± 0.3 | ND | |
kcat/KM (μM−1 s−1) | 0.36 ± 0.004 | 4.24 ± 0.076 | 0.086 ± 0.002 | |
K224R | KM (μM) | 179 ± 17 | 3 ± 0.2 | 69 ± 2 |
kcat (s−1) | 294 ± 22 | 39 ± 2 | 62 ± 0.2 | |
kcat/KM (μM−1 s−1) | 1.65 ± 0.04 | 12.81 ± 0.28 | 0.90 ± 0.03 | |
D225E | KM (μM) | 41 ± 9 | 4 ± 0.1 | 50 ± 8 |
kcat (s−1) | 102 ± 0.8 | 38 ± 0.1 | 117 ± 8 | |
kcat/KM (μM−1 s−1) | 2.54 ± 0.56 | 9.78 ± 0.17 | 2.37 ± 0.24 | |
G232A | KM (μM) | 178 ± 40 | 24 ± 2 | ND |
kcat (s−1) | 417 ± 24 | 90 ± 1 | ND | |
kcat/KM (μM−1 s−1) | 2.39 ± 0.41 | 3.80 ± 0.25 | 0.63 ± 0.008 | |
N233H | KM (μM) | 56 ± 3 | 3 ± 0.5 | 134 ± 43 |
kcat (s−1) | 591 ± 6 | 55 ± 3 | 286 ± 30 | |
kcat/KM (μM−1 s−1) | 10.58 ± 0.61 | 20.42 ± 2.83 | 2.22 ± 0.49 | |
N233Q | KM (μM) | 160 ± 13 | 1.1 ± 0.3 | 565 ± 84 |
kcat (s−1) | 279 ± 23 | 25 ± 1 | 288 ± 31 | |
kcat/KM (μM−1 s−1) | 1.74 ± 0.01 | 22.95 ± 6.85 | 0.51 ± 0.02 |
ND not determined
aData are mean and standard deviations of at least two independent experiments