Table 2.
The kinetic parameters of the purified BdCAD3 and BdCAD5 with coniferyl aldehyde and coniferyl alcohol as substrates
| Enzyme | Substrate | Km (µM) | Vmax (nkat mg–1) | kcat (s–1) | kcat Km –1 (s–1 µM–1) |
|---|---|---|---|---|---|
| BdCAD3 | Coniferyl aldehyde | 33.3±3.80 | 154.3±6.86 | 8.66 | 0.26 |
| NADPH | 1.30±0.12 | 114.9±2.04 | 6.45 | 4.96 | |
| Coniferyl alcohol* | 509.1±134.0 | 611.5±72.1 | 34.33 | 0.067 | |
| NADP+* | 66.0±11.79 | 93.3±3.31 | 5.24 | 0.079 | |
| BdCAD5 | Coniferyl aldehyde | 3.1±0.44 | 492.3±19.16 | 38.60 | 12.87 |
| NADPH | 7.2±1.7 | 487.1±32.1 | 38.19 | 5.30 | |
| Coniferyl alcohol* | 1.44±0.24 | 131.8±4.55 | 10.35 | 7.19 | |
| NADP+ | 0.72±0.10 | 125.9±3.55 | 9.87 | 13.71 |
The parameters were calculated by fitting the Michaelis–Menten equation on initial rate experimental data using non-linear fitting (n = 3; except *, n = 1) using OriginPro (Originlab). Reactions were carried out at 23 °C and the protein concentrations were 0.42 (BdCAD5) and 0.58 µg/ml (BdCAD3).