TABLE 1.
Purification of TodC1 from JM109(pDTG626)a
| Purification step | Total protein (mg) | Activityb (U) | Sp act (U/mg of TodC1) | Recovery (%) | Purifi-cation (fold) |
|---|---|---|---|---|---|
| Crude cell extract | 2,893 | 554 | 0.19 | 100 | 1 |
| Q-Sepharose | 378 | 196 | 0.52 | 35 | 2.7 |
| Hydroxyapatite | 126 | 310 | 2.46 | 56 | 13 |
a
TDO activity was determined by measuring the formation of radioactive cis-toluene dihydrodiol (32). Purified TodC1 (20), TodB, and TodA (19) were used to assay pooled fractions from each column. Protein concentrations were determined by the method of Bradford (2) with bovine serum albumin as the standard.
b
One unit of activity was defined as the amount of protein required to form 1 μmol of cis-toluene dihydrodiol per min.