Cloned (Comment) | Organism |
---|---|
nitrile hydratase from Comamonas testosteroni is expressed in Escherichia coli, strain TG1, overexpressed chaperonin system | Comamonas testosteroni |
Protein Variants | Comment | Organism |
---|---|---|
additional information | Fe of an Fe-dependent recombinant nitrile hydratase (wild-type, NilFe) is replaced by Co generating an Co-substituted enzyme (NilCo) | Comamonas testosteroni |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-Nitrophenol | 10 microM, 40-50% residual activity of NilFe, 90% residual activity of NilCo | Comamonas testosteroni | |
3-Nitrophenol | 10 microM, 40-50% residual activity of NilFe, fully active NilCo | Comamonas testosteroni | |
4-nitrophenol | 10 microM, 40-50% residual activity of NilFe, 80% residual activity of NilCo | Comamonas testosteroni | |
7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole | - |
Comamonas testosteroni | |
acrylonitrile | - |
Comamonas testosteroni | |
Butyric acid | - |
Comamonas testosteroni | |
CO | inhibition of NilCo, reversible by photoactivation, no inhibition of NilFe | Comamonas testosteroni | |
KCN | - |
Comamonas testosteroni | |
NaNO2 | NilFe is unaffected by NaNO2 | Comamonas testosteroni | |
NO | inhibition of NilCo and NilFe, reversible by photoactivation with 150 W white light | Comamonas testosteroni | |
tertiary-butyl isonitrile | NilFe is unaffected by tertiary-butyl isonitrile | Comamonas testosteroni |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
(25R)-3beta-hydroxycholest-5-en-27-oate | +/- 0.04, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilFe | Comamonas testosteroni | |
0.19 | - |
(25R)-3beta-hydroxycholest-5-en-27-oate | +/- 0.06, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilCo | Comamonas testosteroni |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
22700 | - |
alpha2beta2, 2 * 22800 + 2 * 22700 | Comamonas testosteroni |
22800 | - |
alpha2beta2, 2 * 22800 + 2 * 22700 | Comamonas testosteroni |
90000 | - |
gel filtration | Comamonas testosteroni |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Comamonas testosteroni | - |
Fe-dependent, wild-type nitrile hydratase is compared with artificial, Co-dependent nitrile hydratase | - |
Purification (Comment) | Organism |
---|---|
CoCl2, an overexpressed chaperonin system and butyric acid is necessary for fully soluble and active NilCo protein, 95% purity on 12% SDS-PAGE | Comamonas testosteroni |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
vmax: 28 +/- 5 nmol/min/mg, NilCo | Comamonas testosteroni |
additional information | - |
vmax: 30 +/- 5 nmol/min/mg, NilFe | Comamonas testosteroni |
0.018 | - |
0.5 mM NaNO2, NilFe | Comamonas testosteroni |
15 | - |
+/- 5, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilCo | Comamonas testosteroni |
15 | - |
+/- 5, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilFe | Comamonas testosteroni |
100 | - |
+/- 15, substrate: 5-cyanovaleric acid, NilFe | Comamonas testosteroni |
110 | - |
+/- 10, substrate: 5-cyanovaleric acid, NilCo | Comamonas testosteroni |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(25R)-3beta-hydroxycholest-5-en-27-oate | - |
Comamonas testosteroni | ? | - |
? | |
5-cyanovaleric acid + H2O | NilFe and NilCo | Comamonas testosteroni | 6-amino-6-oxohexanoic acid | - |
r |
Subunits | Comment | Organism |
---|---|---|
tetramer | alpha2beta2, 2 * 22800 + 2 * 22700 | Comamonas testosteroni |
Synonyms | Comment | Organism |
---|---|---|
NHase | - |
Comamonas testosteroni |
NilCo | - |
Comamonas testosteroni |
NilFe | - |
Comamonas testosteroni |
nitrile hydratase | - |
Comamonas testosteroni |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.9 | 7.2 | NilFe, NilCo | Comamonas testosteroni |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole | +/- 0.0005, NilFe | Comamonas testosteroni | |
0.002 | - |
7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole | +/- 0.0008, NilCo | Comamonas testosteroni | |
0.0025 | - |
KCN | +/- 0.001, NilCo | Comamonas testosteroni | |
0.003 | - |
NaNO2 | +/- 0.0005, NilCo | Comamonas testosteroni | |
0.003 | - |
KCN | +/- 0.0015, NilFe | Comamonas testosteroni | |
0.004 | - |
tertiary-butyl isonitrile | +/- 0.001, NilCo | Comamonas testosteroni | |
0.005 | - |
acrylonitrile | +/- 0.0005, NilCo | Comamonas testosteroni | |
0.007 | - |
acrylonitrile | +/- 0.001, NilFe | Comamonas testosteroni |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.003 | - |
+/- 0.001, NilCo | Comamonas testosteroni | KCN | |
0.003 | - |
+/- 0.001, NilCo | Comamonas testosteroni | 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole | |
0.004 | - |
+/- 0.001, NilFe | Comamonas testosteroni | 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole | |
0.005 | - |
+/- 0.001, NilFe | Comamonas testosteroni | KCN | |
0.005 | - |
+/- 0.001, NilFe | Comamonas testosteroni | 2-Nitrophenol | |
0.005 | - |
+/- 0.003, NilCo | Comamonas testosteroni | acrylonitrile | |
0.007 | - |
+/- 0.001, NilCo | Comamonas testosteroni | tertiary-butyl isonitrile | |
0.008 | - |
+/- 0.001, NilCo | Comamonas testosteroni | NaNO2 | |
0.0086 | - |
pH 7.9, NilCo | Comamonas testosteroni | 4-nitrophenol | |
0.009 | - |
+/- 0.002, NilFe | Comamonas testosteroni | acrylonitrile | |
0.01 | - |
+/- 0.002, NilFe | Comamonas testosteroni | 4-nitrophenol | |
0.01 | - |
+/- 0.002, NilFe | Comamonas testosteroni | 3-Nitrophenol | |
0.1 | - |
value above, NilFe | Comamonas testosteroni | tertiary-butyl isonitrile | |
0.3 | - |
+/- 0.1, NilCo | Comamonas testosteroni | Butyric acid | |
0.4 | - |
+/- 0.1, NilFe | Comamonas testosteroni | Butyric acid | |
0.5 | - |
value above, NilFe | Comamonas testosteroni | NaNO2 |