TABLE 1.
Purification of recombinant HexA and M6PHexA
| Step no. | Purification method | Protein (mg) | Total activity (mmol/h)
|
Sp act (mmol/h/mg)
|
Recovery of MUG (%) | Purification (fold) of MUG | MUG/MUGS ratio | ||
|---|---|---|---|---|---|---|---|---|---|
| MUG | MUGS | MUG | MUGS | ||||||
| 1 | Culture supernatant, 4 liters | 1,500 | 74 | 43 | 0.051 | 0.029 | 100 (23)a | 1.0 | 1.7 |
| 2 | HisTrap (HexA) | 7.6 | 11 | 4.3 | 1.4 | 0.57 | 14 | 28 | 2.6 |
| 3 | Mannosidase treatment | 16 | 7.9 | 3.2 | 0.51 | 0.20 | 11 | 10 | 2.5 |
| 4 | HisTrap (M6PHexA) | 3.3 | 4.9 | 1.7 | 1.5 | 0.50 | 6.6 | 29 | 2.9 |
a
Amount of HexA in the total isozymes, calculated from their ratio as detected by DEAE elution. Steps 2 and 4 describe purified HexA and M6PHexA, respectively.