Table 3.
Substrate Kinetics
Enzyme | Substrate | KM (µM) | kcat (s−1) | kcat/KM (µM−1 s−1) | Relative Catalytic Eff.a |
---|---|---|---|---|---|
Wild-type | Acetate | 6047 ± 1024 | 276.8 ± 21.4 | 0.045 ± 0.008 | |
Propionate | 9413 ± 1709 | 261.0 ± 20.8 | 0.027 ± 0.005 | ||
Glycine | 9450 ± 1658 | 259.4 ± 13.3 | 0.027 ± 0.005 | ||
V386A | Acetate | 6403 ± 701 | 183.5 ± 10.3 | 0.028 ± 0.003 | 0.6 |
Propionate | 1539 ± 119 | 93.4 ± 2.4 | 0.060 ± 0.005 | 2.2 | |
V310D | Acetate | 2464 ± 321 | 232.0 ± 12.3 | 0.090 ± 0.010 | 2.0 |
Glycine | 8367 ± 753 | 165.5 ± 6.4 | 0.020 ± 0.002 | 0.7 |
Relative catalytic efficiency is defined as the ratio of kcat/KM for a particular substrate for the mutant enzyme compared to the wild-type.