TABLE 4.
Relative activities of PhnI and PhnII in recombinant cells depending on coexpression of PhnA3 and PhnA4
| Expression system | Gene product(s) | Hydroxylase activity with:
|
|
|---|---|---|---|
| Phenanthrenea | Salicylateb | ||
| BL21(DE3)(pSD9) | PhnI | 2.8 | —c |
| BL21(DE3)(pSD9)(pEB432) | PhnI + PhnA4 | 14.8 | — |
| BL21(DE3)(pSD9)(pEB431) | PhnI + PhnA4 + PhnA3 | 100 | — |
| KT2442(pSD9) | PhnI | 28.7 | — |
| BL21(DE3)(pVDE11) | PhnII | — | 4.6 |
| BL21(DE3)(pVDE11)(pEB432) | PhnII + PhnA4 | — | 8.5 |
| BL21(DE3)(pVDE11)(pEBA3) | PhnII + PhnA3 | — | 34.8 |
| BL21(DE3)(pVDE11)(pEB431) | PhnII + PhnA4 + PhnA3 | NDd | 100 |
a
Determined by GC-MS analysis of the diol product formed after 6 h of incubation. The values are the relative GC-MS peak areas normalized to 100% maximal activity.
b
Calculated from HPLC measurements and expressed as percentages of the maximal activity (100% was 0.24 μmol of catechol/h/mg of protein).
c
—, not tested.
d
ND, no product detected.