TABLE 2.
Data from purification of StyB from recombinant E. coli JM109 (and wild-type Pseudomonas strain VLB120)
| Purification step | Total protein (mg) | Sp acta (U mg−1) | Total activity (U) | Purification factor |
|---|---|---|---|---|
| Preparation of crude extract | 200 (780)b | NDc (0.3) | NDc (234) | NDc (1) |
| IB solubilizationd | 40 | 23 | 932 | 1 |
| Refoldingd | 20 | 197 | 3,930 | 8.5 |
| Pooling after IECe | 15 (61.2) | 200 (2) | 3,000 (104) | 8.7 (6.6) |
a
Activity was determined by measuring the NADH oxidation rate as described in Materials and Methods.
b
The purification utilized 2.3 (6) g (wet weight) of cells.
c
ND, Not determined because StyB was in inclusion body form and therefore not active.
d
Pseudomomas strain VLB120 did not form inclusion bodies (IB); therefore; no data are given for wild-type StyB.
e
IEC, ion-exchange chromatography.