TABLE 1.
Partial purification of NADH:menadione oxidoreductase from S. wolfei
| Purification step | Protein amt (mg) | Sp act (U/mg)a
|
Yield (%) | Purification factor | |
|---|---|---|---|---|---|
| NADH oxidationb | NADH:menadione oxidoreductasec | ||||
| First preparation run | |||||
| Cell extract | 63.9 | 0.3 | 1.7 | NAd | NA |
| Soluble fraction | 41.4 | 0.3 | 3.4 | 100 | 1 |
| DEAE-Sepharose CL-4B (pH 8.0) | 10.2 | 0.56 | 9.9 | 72 | 2.9 |
| MonoQ (pH 8.0) | 1.07 | 2.49 | 19.9 | 15 | 5.8 |
| POROS HQ/H (pH 8.0) | 0.6 | 1.65 | 16.5 | 7 | 4.8 |
| MonoQ (pH 7.5) | 0.04 | 2.34 | 59.3 | 1.6 | 17.4 |
| Second preparation run | |||||
| Cell extract | 72 | 1.7 | 5.9 | NA | NA |
| Soluble fraction | 45.4 | 3.0 | 4.4 | NA | NA |
| Membrane fraction | 28.8 | 0.3 | 1.55 | 100 | 1 |
| Solubilized membrane protein | 14.16 | 0.37 | 5.14 | 163 | 3.3 |
| DEAE-Sepharose CL-6B (pH 8.0) | 7.92 | 0.2 | 4.75 | 84.3 | 3.06 |
| MonoQ (pH 8.0) | 0.88 | 1.46 | 20.2 | 39.8 | 13.03 |
| MonoQ (pH 7.5) | 0.48 | 0.66 | 26.46 | 12.7 | 17.07 |
a
Mean activities of three independent measurements are shown.
b
Unspecific oxidation of NADH before the addition of menadione.
c
Values corrected for unspecific NADH-oxidizing activity.
d
NA, not applicable.