TABLE 4.
Purification of hydroxylase and reductase of sMMO from Methylocystis sp. strain WI 14
| sMMO component | Purification step | Total amt of protein (mg) | Total activity (U) | Sp act (mU/mg) | Purification (fold) | Yield (%) |
|---|---|---|---|---|---|---|
| Hydroxylase (component A) | Ultracentrifuged extract | 487 | 24 | 49 | 1 | 100 |
| DEAE-Sepharose (0.15 M NaCl, step) | 113 | 19 | 167 | 3.4 | 79 | |
| DEAE-Sepharose (0–0.15 M NaCl, linear) | 34 | 15 | 439 | 8.9 | 64 | |
| Superose 12 HR | 10 | 5 | 518 | 10.5 | 21 | |
| Reductase (component C) | Ultracentrifuged extract | 75.1 | 11.3 | 0.15 | 1 | 100 |
| DEAE-Sepharose (0.3–0.5 M NaCl, step) | 6.8 | 12.3 | 1.82 | 12 | 109 | |
| DEAE-Sepharose (0.3–0.5 M NaCl, linear) | 0.36 | 8.5 | 23.86 | 158 | 75 | |
| Blue-Sepharose | 0.11 | 3.3 | 30.64 | 204 | 29 |