Crystallization (Comment) | Organism |
---|---|
x-ray diffraction, special focus on iron center vicinity and hydrogen bond network, similar structures of wild type and alphaQ90N mutant enzyme in nitrosylated state, resolution: 1.43 A | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
Q90E | alpha subunit | Rhodococcus sp. |
Q90N | alpha subunit | Rhodococcus sp. |
R56K | beta subunit | Rhodococcus sp. |
General Stability | Organism |
---|---|
purified mutant NHases are stored in the dark without n-butyric acid, before use, NHases are activated by light irradiation. | Rhodococcus sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
n-butyric acid | - |
Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.28 | - |
Methacrylonitrile | mutant alphaQ90E, vmax: 280 micromol/min/mg | Rhodococcus sp. | |
0.68 | - |
Methacrylonitrile | wild type, vmax: 1200 micromol/min/mg | Rhodococcus sp. | |
1.9 | - |
Methacrylonitrile | mutant betaR56K, vmax: 2.5 micromol/min/mg | Rhodococcus sp. | |
2.6 | - |
Methacrylonitrile | mutant alphaQ90N, vmax: 57 micromol/min/mg | Rhodococcus sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | - |
- |
- |
Rhodococcus sp. N771 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
3 successive column chromatographies in the presence of 40 mM n-butyric acid (only wild type), alphaQ90N mutein is purified in nitrosylated state in the dark | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
methacrylonitrile + H2O | - |
Rhodococcus sp. | methacrylamide | - |
? | |
methacrylonitrile + H2O | - |
Rhodococcus sp. N771 | methacrylamide | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NHase | - |
Rhodococcus sp. |
nitrile hydratase | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
120 | - |
Methacrylonitrile | mutant betaR56K | Rhodococcus sp. | |
2600 | - |
Methacrylonitrile | mutant alphaQ90N | Rhodococcus sp. | |
13000 | - |
Methacrylonitrile | mutant alphaQ90E | Rhodococcus sp. | |
55000 | - |
Methacrylonitrile | wild type | Rhodococcus sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
2.6 | - |
Butyric acid | +/- 0.2, substrate: methacrylonitrile, mutant betaR56K | Rhodococcus sp. | |
5.1 | - |
Butyric acid | +/- 0.3, substrate: methacrylonitrile, mutant alphaQ90E | Rhodococcus sp. | |
6.2 | - |
Butyric acid | +/- 0.9, substrate: methacrylonitrile, wild type | Rhodococcus sp. | |
14 | - |
Butyric acid | +/- 2.7, substrate: methacrylonitrile, mutant alphaQ90N | Rhodococcus sp. |