EC Number   |
Protein Variants |
Reference |
|---|
   1.6.5.10 | D50N |
less than 0.1% of wild-type activity |
714140 |
| 1.6.5.10 | H117A |
less than 0.1% of wild-type activity |
714140 |
| 1.6.5.10 | K84M |
complete loss of activity |
714140 |
| 1.6.5.10 | K84R |
complete loss of activity |
714140 |
| 1.6.5.10 | more |
improvement of the kinetic and thermodynamic stability of the azoreductase by directed evolution via rational design approaches, five rounds of mutagenesis/recombination are followed by high-throughput screening. Mutant 1B6 shows a 300fold higher half-life at 50°C compared to the wild-type enzyme. mutant 1B6 has a folded state slightly less stable than the wild-type (with lower melting and optimal temperatures) but in contrast is more resistant to irreversible denaturation. The superior kinetic stability of 1B6 variant is therefore related to an increased resistance of the unfolded monomers to aggregation through the introduction of mutations that disturb hydrophobic patches and increase the surface net charge of the protein. Mutants 2A1 and 2A1-Y179H show increased thermodynamic stability with a 10-20°C higher melting temperature than wild-type, these residues are mostly involved in strengthening the solvent-exposed loops or the inter-dimer interactions of the folded state. Molecular details of mutations that improve stability, overview |
-, 743650 |
| 1.6.5.10 | P187S |
Pro187Ser polymorphism in NQO1 has a limited role in the development of Tardive dyskinesia (a potentially irreversible side effect of antipsychotic medication treatment that occurs in approximately 25% of chronically treated schizophrenia patients) |
701035 |
| 1.6.5.10 | Q192R |
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 60 min) relative to parental mutant variant B1G6 |
-, 743650 |
| 1.6.5.10 | Q192R/A46P/V159A |
site-directed mutagenesis, analysis of initial activity and thermostability (at 55°C, 90 min) relative to parental mutant variant 16B7 |
-, 743650 |
| 1.6.5.10 | Q192R/A46P/V159A/A48P |
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 2A1 |
743650 |
| 1.6.5.10 | Q192R/A46P/V159A/C129S |
site-directed mutagenesis, analysis of initial activity and thermostability (at 60°C, 45 min) relative to parental mutant variant 23C10 |
743650 |
