Table 1. Quantification by NMR of products formed in engineered yeast strains.
| Expressed enzymes | Product | Concentration (μmol l−1) | Concentration (mg l−1) | Ratio product/miltiradiene | Ratio product/ferruginol |
|---|---|---|---|---|---|
| Empty vector control | ND | − | − | ||
| GGPPS:CPS:MiS | Miltiradiene | 5.86±1.52 | 1.59±0.41 | 1 | |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 0.93±0.18 | 0.25±0.05 | 1 | 0.15 |
| CYP76AH1 | Ferruginol | 6.37±0.55 | 1.82±0.16 | 6.85 | 1 |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 0.86±0.22 | 0.23±0.06 | 1 | 3.74 |
| CYP76AH22 | Ferruginol | 0.23±0.05 | 0.07±0.01 | 0.27 | 1 |
| 11-hydroxyferruginol | 4.18±0.48 | 1.26±0.14 | 4.86 | 18.17 | |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 1.15±0.11 | 0.31±0.03 | 1 | 2.80 |
| CYP76AH1D301E,N303S,V479F | Ferruginol | 0.41±0.09 | 0.12±0.03 | 0.35 | 1 |
| 11-hydroxyferruginol | 7.43±1.20 | 2.25±0.36 | 6.46 | 18.12 | |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 3.72±0.25 | 1.01±0.07 | 1 | 0.558 |
| CYP76AH22E301D,S303N,F478V | Ferruginol | 6.78±0.20 | 1.94±0.06 | 1.82 | 1 |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 0.73±0.14 | 0.20±0.04 | 1 | 1.87 |
| CYP76AH22:CYP76AK8 | Ferruginol | 0.39±0.09 | 0.11±0.03 | 0.53 | 1 |
| Carnosic acid | 8.26±1.64 | 2.74±0.55 | 11.31 | 21.18 | |
| GGPPS:CPS:MiS:ATR1: | Miltiradiene | 2.41±0.25 | 0.66±0.07 | 1 | 3.60 |
| CYP76AH1:CYP76AK8 | Ferruginol | 0.67±0.10 | 0.19±0.03 | 0.28 | 1 |
| Pisiferal | 3.03±0.84 | 0.91±0.25 | 1.25 | 4.52 | |
| Pisiferic acid | 1.82±0.75 | 0.57±0.24 | 0.76 | 2.72 |
CPS, copalyl diphosphate synthase; GGPPS, geranylgeranyl diphosphate synthase; MS, miltiradiene synthase; ND, not detected.
The concentrations of products, which could be detected with a signal sufficient for quantification, are given with their s.d. (n=3). The concentrations are normalized to the cell density as measured by OD600. The ratios product/miltiradiene or product/ferruginol were calculated with the molar concentrations.