TABLE 2.
Purification of 3-hydroxypropionyl-CoA synthetase from 10 g (wet cell mass) of autotrophically grown Metallosphaera sedula
| Purification step | Volume (ml) | Total activity (μmol min−1)a | Total protein (mg) | Specific activity (μmol min−1 mg−1) | Recovery (%) | Purification (fold) |
|---|---|---|---|---|---|---|
| Cell extract | 31 | 150 | 810 | 0.19 | (100)b | (1) |
| 30% (NH4)2SO4 | 25 | 108 | 550 | 0.20 | 72 | 1.1 |
| Phenyl-Sepharose | 35 | 60 | 160 | 0.38 | 40 | 2.0 |
| MonoQ | 12 | 42 | 37 | 1.1 | 28 | 5.8 |
| Reactive Green | 1.5 | 31 | 4.7 | 6.6 | 21 | 35 |
| Gel Filtration | 1.1 | 16 | 0.9 | 18 | 11 | 95 |
a
Activity was determined at 65°C using the discontinuous assay.
b
Parentheses around entries in row 1 signify set values.