Table 2.
Specific activities and metal contents of urease samples enriched from cells containing selected KaUreD variantsa
| KaUreD variant | Specific activity U/mg protein (% WT) | Ni per UreABC (% WT) | Zn per UreABC | Fe per UreABC |
|---|---|---|---|---|
| WT(-Ni) pKK17D | 0 | 0.004 (4 × 10−5) | 0.13 | 0.27 |
| WT pKKDG | 1690 (100) | 1.1 (100) | 0 | 0.02 |
| D63A pKKDG | 434 (25.7) | 0.54 (49.1) | 0.16 | 0.21 |
| D63Q pKKDG | 55.7 (3.29) | 0.22 (20.0) | 0.34 | 0.07 |
| S85K pKKDG | 487 (28.8) | 0.39 (35.5) | 0.06 | 0.30 |
| D142A pKKDG | 14.1 (0.84) | 0.07 (6.4) | 0.52 | 0.14 |
| E176A pKKDG | 1080 (63.7) | 0.51 (46.4) | 0.09 | 0.11 |
| E176Q pKKDG | 375 (22.2) | 0.43 (39.1) | 0.03 | 0.13 |
| R211A pKKDG | 1230 (72.4) | 0.76 (69.1) | 0.21 | 0.29 |
The urease samples were enriched from cell-free extracts of E. coli pKKD*G grown in LB containing 1 mM NiCl2, except for a sample of urease apoprotein that was obtained from E. coli pKK17D grown in LB lacking nickel (-Ni). Units of urease activity are μmole min−1 (mg protein)−1. Metal contents were determined by inductively coupled plasma-atomic emission spectroscopy.