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Kinetic parameters of the wild-type and engineered acetoacetyl-CoA reductases (PhaBs)a
| PhaB | kcat (s−1) | Km(NADPH) (μM) | Km(AcAcCoA) (μM) | kcat/Km(NADPH) (M−1 s−1) | kcat/Km(AcAcCoA) (M−1 s−1) |
|---|---|---|---|---|---|
| Wild type | 102 | 149 | 5.7 | 6.85 × 105 | 1.80 × 107 |
| Q47L | 249 | 289 | 15.9 | 8.62 × 105 | 1.52 × 107 |
| T173S | 361 | 617 | 13.6 | 5.85 × 105 | 2.65 × 107 |
The activity was measured using His tag fusion proteins in 125 mM Tris-HCl buffer (pH 8.0) at 30°C.
