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Steady state kinetics of mutant enzymes with shallower and narrower cavity.
| Enzymes | wt-hALOX12 | A417I | V418M | A417I/V418M | A417I/V418M/S594T |
|---|---|---|---|---|---|
| KM (µM) | 1.9 ± 0.2 | 0.86 ± 0.1 | 3.2 ± 0.6 | 0.56 ± 0.1 | 0.52 ± 0.1 |
| kcat (s−1) | 28 ± 0.9 | 15 ± 0.4 | 20 ± 2 | 34 ± 1 | 2.7 ± 0.09 |
| kcat/KM (µM)−1s−1 | 14 ± 0.9 | 18 ± 2 | 6.3 ± 0.7 | 59 ± 10 | 5.1 ± 0.9 |
| Product profile (12-HETE:15-HETE:11-HETE) | 100:0:0 | 85:15:0 | 100:0:0 | 75.5:24:0.5 | 56:46:0 |
