Table 1.

Purification of the ISPNAR protein fromRhodococcus sp. strain NCIMB12038

Purification stageaProtein (mg)Activity (U)bSp act (U · mg−1)Recovery (%)
(i)  Cell extract198144.70.731100
(ii)  Blue Sepharose column137113.80.83179
(iii)  Q-Sepharose column23119.65.225c 82
(iv)  Superose 12 column1377.45.97153
  • a See text for details.

  • b One unit represents production of 1.0 nmol ofcis-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.