Table 1.
Steps | Total protein (mg) | A280/A456 | Total activity (nmol H2O2/min) | Yield (%) | Specific activity (nmol/min·mg) | Fold purification |
---|---|---|---|---|---|---|
Serum (500 mL diluted 5-fold) | 31350 | 140 | 4148 | 100 | 0.132 | 1 |
Cation exchange (salt gradient) | 10.3 | 31 | 1686 | 411 | 164 | 1240 |
Hydrophobic interaction | 1.52 | 22 | 560 | 13.5 | 369 | 2793 |
Cation exchange (pH gradient) | 0.57 | 12 | 402 | 10 | 706 | 5345 |
During loading the 2.5 L of diluted serum, the cation exchange column was intentionally overloaded as described in Methods. Without such overloading 82% of the sulfhydryl oxidase applied to the column could be recovered from the salt gradient in a single peak of enzyme activity.