TABLE 1.
Substrate binding, steady state turnover activity with a ArR:Arx:CYP concentration ratio of 1:10:1 (0.5 μm CYP enzyme, 50 mm Tris, pH 7.4), and coupling for the five N. aromaticivorans CYP enzymes with their respective substrates
Coupling is the percentage efficiency of NADH utilization for the formation of products. Data for the assays with 200 mm KCl present are given in parentheses. Rates are reported as mean ± S.D. (n ≥ 3) and given in nmol(nmol CYP)−1 s−1. The NADH consumption rate, product formation rate, and coupling of CYP101D2 with camphor in the presence of 200 mm NH4Cl were 13.5 ± 1.0, 12.6 ± 0.2, and 93%, respectively.
| P450 enzyme/substrate | HS heme | Kd | NADH consumption rate | Product formation rate | Coupling |
|---|---|---|---|---|---|
| % | μm | ||||
| CYP101D2/camphor | 30% (40%) | 5.2 ± 0.7 (3.1 ± 0.5) | 33.7 ± 1.3 (12.4 ± 0.8) | 33.5 ± 1.7 (12.4 ± 0.5) | 99% (99%) |
| CYP101D1/camphor | 30% (40%) | 9.1 ± 1.1 (5.9 ± 0.5) | 30.8 ± 0.6 (7.2 ± 0.03) | 29.3 ± 0.6 (6.8 ± 0.08) | 95% (90%) |
| CYP101D1/2-adamantanone | 50% (60%) | 9.5 ± 1.0 (4.8 ± 0.4) | 27.8 ± 0.6 (7.1 ± 0.3) | 24.0 ± 1.1 (6.4 ± 0.2) | 86% (90%) |
| CYP101B1/β-ionone | ≥95% (≥95%) | 0.23 ± 0.1 (0.09 ± 0.03) | 26.7 ± 1.6 (3.5 ± 0.1) | 16.8 ± 1.0 (2.4 ± 0.03) | 63% (69%) |
| CYP101C1/β-ionone | 20% (20%) | 26.6 ± 4.0 (23.7 ± 1.6) | 44.2 ± 1.3 (7.2 ± 0.2) | 33.7 ± 1.4 (4.7 ± 0.4) | 76% (66%) |
| CYP111A2/linalool | ≥95% (≥95%) | 0.47 ± 0.1 (0.26 ± 0.04) | 48.3 ± 3.3 (11.3 ± 0.6) | 30.7 ± 2.1 (7.7 ± 0.5) | 63% (68%) |