Table 2.
Purification of the PAL enzyme from Trichosporon cutaneum a.
| Method | Total activityb (U) | Total protein (mg) | Specific activity (U/mg) | Purification fold | Yield (%) |
|---|---|---|---|---|---|
| Anion exchange | |||||
| Crude | 0.05 ± 0.01 | 0.38 ± 0.09 | 0.15 ± 0.00 | — | 100 |
| Post | 0.01 ± 0.00 | 0.001 ± 0.00 | 7.39 ± 0.73 | 49.6 | 20 |
| Acid precipitation | |||||
| Crude | 0.16 ± 0.03 | 1.04 ± 0.74 | 0.20 ± 0.10 | — | 100 |
| Post | 0.15 ± 0.04 | 0.61 ± 0.28 | 0.26 ± 0.06 | 1.3 | 94 |
| Aqueous two-phasec | |||||
| Crude | 0.07 ± 0.03 | 0.39 ± 0.14 | 0.21 ± 0.05 | — | 100 |
| PEG/Na2SO4 | 0.02 ± 0.00 | 0.08 ± 0.00 | 0.33 ± 0.03 | 1.5 | 28 |
| PEG/(NH4)2SO4 | 0.02 ± 0.00 | 0.10 ± 0.08 | 0.10 ± 0.04 | 0.5 | 28 |
| PEG/ Na2CO3 | 0.01 ± 0.00 | 0.31 ± 0.04 | 0.02 ± 0.01 | 0.1 | 14 |
aResults are the average ± range of two independent trials. Crude samples are cell culture extracts obtained from a PD-10 column. b1 U of activity = 1 μmoL of released tran-cinnamic acid per min. cFor aqueous two-phase partitioning, the same crude enzyme extraction was used for each of the PEG/salt systems.