EC Number   |
Protein Variants |
Reference |
|---|
   1.5.3.5 | D166Q |
mutation slightly reduces the KM for nicotine, it also reduced enzyme turnover by 5fold with nicotine |
764029 |
| 1.5.3.5 | K287M |
mutation results in an about 10-fold decreases in kcat/Km and k(red) for (S)-6-hydroxynicotine and a 6000-fold decrease in the kcat/Km value for oxygen |
741943 |
| 1.5.3.5 | more |
adding a maltose-binding protein tag onto the N-terminus markedly increases the thermal stability of the enzyme and increases the observed Vmax value for 6-OH-nicotine by about 4.5fold, due to an increase in the occupancy of the flavin cofactor following expression and purification |
764029 |
| 1.5.3.5 | N166A |
mutation results in an about 30fold decrease in kcat/Km and k(red) for (S)-6-hydroxynicotine, respectively, with larger effects on the kcat/Km value for (S)-6-hydroxynornicotine. The shapes of the pH profiles are not altered |
741943 |
| 1.5.3.5 | N462H |
the variant shows moderately higher oxidase activity, reductive half-reaction using (S)-nicotine is significantly slower than that of wild-type enzyme |
-, 764183 |
| 1.5.3.5 | N462Y/W427Y |
mutant enzyme shows increased catalytic activity |
-, 764183 |
| 1.5.3.5 | R274A/Y311W/C417W |
combination of mutations predicted to enhance enzyme stability, and mutation Y311W. The triple mutant displays an increased kcat value for nicotine resulting in a comparatively robust oxidation of (S)-nicotine, at the same time reducing the specificity for (S)-OH-nicotine by more than 100fold and increasing that for (S)-nicotine by more than fold |
764029 |
| 1.5.3.5 | W427V |
the variant shows moderately higher oxidase activity, reductive half-reaction using (S)-nicotine is significantly slower than that of wild-type enzyme |
-, 764183 |
| 1.5.3.5 | Y311F |
mutation results in an about 30fold decrease in kcat/Km and k(red) for (S)-6-hydroxynicotine, respectively, with larger effects on the kcat/Km value for (S)-6-hydroxynornicotine. The shapes of the pH profiles are not altered |
741943 |
| 1.5.3.5 | Y311W |
active site residue Tyr311 forms a hydrogen bond with the hydroxyl group of (S)-6-OH-nicotine within the catalytic pocket. Replacement by a tryptophan residue reduces the kcat for (S)-6-OH-nicotine by more than 6fold |
764029 |
