TABLE 1.
Purification of the ISPNAR protein from Rhodococcus sp. strain NCIMB12038
| Purification stagea | Protein (mg) | Activity (U)b | Sp act (U · mg−1) | Recovery (%) |
|---|---|---|---|---|
| (i) Cell extract | 198 | 144.7 | 0.731 | 100 |
| (ii) Blue Sepharose column | 137 | 113.8 | 0.831 | 79 |
| (iii) Q-Sepharose column | 23 | 119.6 | 5.225c | 82 |
| (iv) Superose 12 column | 13 | 77.4 | 5.971 | 53 |
a
See text for details.
b
One unit represents production of 1.0 nmol of cis-dihydrodiol · min−1.
c
Fractions A and B were both necessary for enzyme activity. When either fraction was assayed alone, coenzyme-dependent specific activity was reduced to approximately 1 and 14%, respectively, of the combined activity. In enzyme assays used for stages iii and iv, 0.3 mg of fraction A and 0.1 mg of fraction B or ISPNAR were combined in 200 μl of assay mixture.