TABLE 1.
Purification of native StoHK from S. tokodaii
| Purification step | Protein (mg) | Activity (U)a | Specific activity (U mg−1)a | Yield (%)b | Purification (fold)b |
|---|---|---|---|---|---|
| Cell extracts | 8,160 | 5.6c | 0.0007c | ||
| DEAE-Sepharose | 1,089 | 11 | 0.010 | 100 | 1 |
| Butyl-Toyopearl | 87 | 6.8 | 0.078 | 62 | 7.8 |
| Mono Q | 2.0 | 3.0 | 1.5 | 27 | 1.5 × 102 |
| Resource PHE | 0.10 | 1.6 | 17 | 15 | 1.7 × 103 |
| Superdex 200 | 0.013 | 0.33 | 26 | 3.1 | 2.6 × 103 |
a
The enzyme activity was measured at 50°C using 5 mM glucose and 2 mM ATP as substrates in the presence of 4 mM Mg2+.
b
Yield and purification were calculated with respect to the DEAE-Sepharose chromatography step.
c
The enzyme activity was determined by subtracting the GDH activity from the value obtained with a G6PDH-coupled assay.