TABLE 1.
Purification of MPLA activity from M. smegmatis
| Purification step for MPLA activity from M. smegmatis 14468 | Vol (ml) | Total proteina (mg) | Total activityb (trace units of FA) | Sp act (trace units of FA mg−1) | % Yield | Purification factor (fold) |
|---|---|---|---|---|---|---|
| Culture supernatant | 7,185 | 11,783 | 23,719,984 | 0.171 | 100 | 1 |
| 60% ammonium sulfate | 63.6 | 91.6 | 5,169,812 | 616 | 22 | 3,602 |
| precipitation | ||||||
| Cation-exchange flowthrough | 108.8 | 97.0 | 4,382,557 | 466 | 18.5 | 2,725 |
| UNO Q6, pH 8.1 | 90.0 | 13.5 | 552,123c | 3,029 | 2.3 | 17,713 |
| UNO Q1, pH 8.6 | 50 | 1.5 | 51,327c | 22,812 | 0.22 | 133,403 |
| UNO Q1, pH 9.2 | 16 | 0.88 | 29,476c | 38,062 | 0.12 | 222,585 |
| Native gel electrophoresis | 0.3 | 0.03 | —d |
a
Total protein was measured using the BCA assay.
b
Total activity was measured using a [14C]PC assay and Quantity One software to measure trace units of released FA.
c
Total activity in these specimens may be inaccurate because all of the substrate was consumed.
d
These calculations were not made because purified preparation was conserved for mass spectrometry and other assays.