TABLE 2.
Substrate specificity of C. formosus diverged desaturases
Shown are fatty acids as % of yeast total fatty acid composition, as determined by GC-MS analysis of FAMEs, in the heterologous expression of Cf0745 and Cf0807 supplemented with linoleic (+18:2 Δ9c,12c, 0.5 mm) or crepenynic acid (+18:1 Δ9c,12a, 0.5 mm). Cultures were grown at 15 °C, 250 rpm in media containing NP-40 (1 mg/ml). Results are expressed as the mean ± sd, n = 3–6. ND, not detected.
| Constructs | Fatty acids |
|||||||
|---|---|---|---|---|---|---|---|---|
| 16:0 | 16:1 | 16:2 Δ9,12 | 18:0 | 18:1 | 18:2 Δ9,12 | 18:1 Δ9c,12a | 18:2 Δ9c,12a,14c | |
| +18:2Δ9,12a | ||||||||
| pYES2 | 13.6 ± 3.1 | 4.00 ± 0.12 | ND | 2.47 ± 0.62 | 2.06 ± 0.16 | 77.9 ± 3.8 | ND | ND |
| H6Cf0745 | 10.3 ± 0.1 | 2.52 ± 0.17 | 0.40 ± 0.02b | 1.81 ± 0.06 | 0.63 ± 0.05 | 84.4 ± 0.2b | ND | ND |
| H6Cf0807 | 12.8 ± 1.4 | 3.75 ± 0.40 | 0.17 ± 0.02c | 2.43 ± 0.38 | 2.11 ± 0.27 | 78.2 ± 2.5c | 0.52 ± 0.08 | ND |
| +18:1Δ9,12aa | ||||||||
| pYES2 | 17.0 ± 0.7 | 21.3 ± 1.5 | ND | 4.98 ± 0.10 | 18.1 ± 1.2 | ND | 38.6 ± 3.3 | ND |
| vH6Cf0745 | 21.1 ± 0.6 | 20.6 ± 1.3 | 1.48 ± 0.29b | 5.75 ± 0.70 | 7.7 ± 1.0 | 8.20 ± 0.30b | 31.9 ± 1.4 | 3.26 ± 0.19 |
| H6Cf0807 | 21.6 ± 0.2 | 21.7 ± 3.6 | 0.96 ± 0.26c | 6.45 ± 0.66 | 15.0 ± 2.0 | 0.43 ± 0.19c | 33.9 ± 4.9 | ND |
| PchFAD2d | 34.8 ± 1.3 | 18.2 ± 1.4 | 1.40 ± 0.22b | 10.7 ± 0.5 | 7.99 ± 0.48 | 4.68 ± 0.15b | 18.7 ± 3.6 | 3.46 ± 0.26 |
a Fatty acid supplemented to expression culture.
b Δ9c,12c.
c Δ9c,12t.
d Expression temperature of 30 °C.