TABLE 1.
Partial purification of acryloyl-CoA reductase from M. sedula and heterologously produced acryloyl-CoA reductase from S. tokodaii
| Enzyme (accession no.) | Purification step | Volume (ml) | Total activity (μmol min−1)a | Protein (mg) | Specific activity (μmol mg−1 min−1) | Yield (%) | Purification (n-fold) |
|---|---|---|---|---|---|---|---|
| M. sedula reductase (Msed_1426) | Cell extract | 82.5 | 8,400 | ||||
| Heat precipitation (85°C) | 45.5 | 43 | 3,000 | 0.014 | 100 | 1 | |
| Dialysis | 43 | 40 | 1,800 | 0.018 | 94 | 2 | |
| Q-Sepharose | 86.5 | 42 | 470 | 0.09 | 98 | 6 | |
| Carboxymethylcellulose | 16.5 | 18 | 9.9 | 1.8 | 43 | 130 | |
| Phenylsepharose | 20 | 4.6 | 1.6 | 2.9 | 11 | 210 | |
| S. tokodaii reductase (ST0480) | Cell extract | 2.6 | 140 | ||||
| Heat precipitation (85°C) | 2 | 46 | 10 | 4.6 | 100 | 1 | |
| His-Trap | 7.8 | 32 | 1.7 | 18.7 | 70 | 4.1 |
a
Activities were determined using the respective photometric assay at 65°C.