EC Number   |
Protein Variants |
Reference |
|---|
  1.2.1.77 | C296A |
same purification behavior as the native enzyme indicating that the mutation does not alter basic enzyme structure, but no catalytic activity detected consistent with a critical nucleophilic role for C296 |
726445 |
| 1.2.1.77 | E167A |
significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 77%, likely candidate to present the catalytic water to the general base E257 that leads to deacylation and product release |
726445 |
| 1.2.1.77 | E257Q |
tolerated with respect to overall protein stability, but dramatic reduction in kcat value and no activity detectable, E257 serves probably as the primary general base to deprotonate the nucleophilic C296 |
726445 |
| 1.2.1.77 | E400A |
91% reduction in kcat value, E400 plays an essential role in facilitating the acquisition of the hydride conformation |
726445 |
| 1.2.1.77 | E496A |
significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 90% |
726445 |
| 1.2.1.77 | H485A |
dramatic reduction in enzyme solubility |
726445 |
| 1.2.1.77 | H485Q |
completely recalcitrant to purification, H485 may be important in stabilizing the nicotinamide amide moiety of NADP+ |
726445 |
| 1.2.1.77 | K168A |
significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 92.5%, K168 likely serves to stabilize charges and tune pKa of the active site glutamate side-chains |
726445 |
