Table 2.
Kinetic parameters of FhCL1, FhCL2 and the mutant enzymes over two different substrates.
| Enzyme | Z-Val-Leu-Lys-AMC | Tos-Gly-Pro-Arg-AMC | ||||
|---|---|---|---|---|---|---|
| kcat (s−1) | KM (μM) | kcat/KM (M−1s−1) | kcat (s−1) | KM (μM) | kcat/KM (M−1s−1) | |
| FhCL1 | 2.32 ± 0.12 | 3.94 ± 0.60 | 5.89 × 105 | 0.035 ± 0.006 | 15.10 ± 2.70 | 2.36 × 103 |
| FhCL1 Leu67Tyr | 0.53 ± 0.11 | 1.72 ± 0.06 | 3.08 × 105 | 0.024 ± 0.003 | 8.52 ± 2.70 | 2.82 × 103 |
| FhCL1 Val157Leu | 0.72 ± 0.07 | 5.23 ± 0.58 | 1.38 × 105 | 0.027 ± 0.001 | 13.16 ± 1.51 | 2.08 × 103 |
| FhCL1 Asn158Thr | 2.12 ± 0.14 | 18.38 ± 4.1 | 1.16 × 105 | 0.023 ± 0.001 | 6.84 ± 1.73 | 3.32 × 103 |
| FhCL1 Leu67Tyr Val157Leu | 0.17 ± 0.010 | 3.85 ± 0.61 | 4.31 × 104 | 0.027 ± 0.003 | 6.57 ± 2.40 | 4.01 × 103 |
| FhCL1 Leu67Tyr Asn158Thr | 1.35 ± 0.10 | 23.30 ± 4.00 | 5.78 × 104 | 0.031 ± 0.004 | 8.81 ± 3.80 | 3.51 × 103 |
| FhCL1 Val157Leu Asn158Thr | 1.67 ± 0.11 | 10.2 ± 1.94 | 1.63 × 105 | 0.015 ± 0.001 | 3.32 ± 1.90 | 4.52 × 103 |
| FhCL1 Leu67Tyr Val157Leu Asn158Thr | 0.65 ± 0.07 | 25.6 ± 6.7 | 2.54 × 104 | 0.030 ± 0.002 | 6.86 ± 1.85 | 4.40 × 103 |
| FhCL2 | 1.65 ± 0.24 | 4.72 ± 1.1 | 3.49 × 105 | 0.720 ± 0.062 | 5.28 ± 2.01 | 1.36 × 105 |