TABLE 2.
Optimization of the lipase/lipase chaperone molar ratio for in vitro refolding
| Chaperone | Molar ratioa | U/mg ofb:
|
|
|---|---|---|---|
| Lipase | Total protein | ||
| Δ70HpHis | 1:0.1 | 520 | 480 |
| 1:0.5 | 3,680 | 2,622 | |
| 1:1 | 3,750 | 2,520 | |
| 1:3 | 4,180 | 1,450 | |
| ompAΔ70HpHis | 1:0.1 | 410 | 380 |
| 1:0.5 | 2,190 | 1,700 | |
| 1:1 | 3,580 | 2,240 | |
| 1:3 | 3,540 | 1,280 | |
a
Defined as the ratio of the amount (in moles) of the denatured mature lipase isolated from inclusion bodies to the amount (in moles) of the denatured chaperone (Δ70HpHis or ompAΔ70HpHis) purified by Ni-NTA chromatography.
b
Specific activity is given as units of lipase per milligram of lipase or milligram of total protein used in the refolding mixture.