TABLE 3.
Purification of β-methylmalyl-CoA lyase from autotrophically grown cells of C. aurantiacusa
| Purification step | Total condensation reaction activity (μmol min−1)b | Total protein (mg) | Specific condensation reaction activity (μmol min−1 mg of protein−1)b | Recovery of condensation reaction activity (%)b | Purification of condensation reaction activity (fold)b | Total l-malyl-CoA cleavage activity (μmol min−1) | Specific l-malyl-CoA cleavage activity (μmol min−1 mg of protein−1) | Recovery of l-malyl-CoA cleavage activity (%) | Purification of l-malyl-CoA cleavage activity (fold) |
|---|---|---|---|---|---|---|---|---|---|
| Cell extractc | 13.1 (18.6) | 768 | 0.017 (0.024) | 100 (100) | 1 (1) | 36.3 | 0.047 | 100 | 1 |
| Heat precipitation (65°C, 10 min) | 14.4 (15.8) | 375 | 0.038 (0.042) | 109 (85.1) | 2.2 (1.7) | ND | ND | ND | ND |
| DEAE-Sepharose | 11.1 (11.6) | 63.9 | 0.174 (0.182) | 84.5 (62.7) | 10.2 (7.5) | ND | ND | ND | ND |
| Phenyl-Sepharose | 5.2 (ND) | 12.6 | 0.414 (ND) | 39.7 (ND) | 24.2 (ND) | ND | ND | ND | ND |
| Gel filtration | 2.1 (ND) | 2.6 | 0.805 (ND) | 16.0 (ND) | 47.1 (ND) | ND | ND | ND | ND |
| Resource Q | 0.92 (0.91) | 1.8 | 0.512 (0.503) | 7.0 (6.9) | 29.9 (21.0) | 2.2 | 1.2 | 6.0 | 26.1 |
a
β-Methylmalyl-CoA lyase was purified from 25 g (wet weight) of C. aurantiacus cells; the glyoxylate reaction was followed at 55°C.
b
The first value refers to the condensation of glyoxylate with propionyl-CoA, and the value in parentheses refers to the condensation of glyoxylate with acetyl-CoA.
c
100,000 × g supernatant.
d
ND, not determined.