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Literature summary for 3.5.5.7 extracted from
- A nitrilase from a metagenomic library acts regioselectively on aliphatic dinitriles (2011), Appl. Microbiol. Biotechnol., 89, 91-98.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | uncultured bacterium |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ag+ | 5% residual activity at 1 mM | uncultured bacterium |  |
| EDTA | 84% residual activity at 1 mM | uncultured bacterium | |
| Fe2+ | 72% residual activity at 1 mM | uncultured bacterium | |
| additional information | Nit1 is not or very weakly affected by Na+, Ca2+, Mg2+, and Fe3+ | uncultured bacterium |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 47 | - |
glutaronitrile | at 30°C in 50 mM sodium phosphate buffer (pH 7.5) and 1 mM dithiothreitol | uncultured bacterium | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 38000 | - |
x * 38000, SDS-PAGE | uncultured bacterium |
| 700000 | - |
gel filtration | uncultured bacterium |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| uncultured bacterium | E1A0Z9 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-NTA column chromatography and Superdex 200 gel filtration | uncultured bacterium |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-methylglutaronitrile + 2 H2O | 77% activity compared to glutaronitrile | uncultured bacterium | 4-cyanopentanoic acid + NH3 | - |
? | |
| 3-cyanopyridine + H2O | 11% activity compared to glutaronitrile | uncultured bacterium | 3-carboxypyridine + NH3 | - |
? | |
| 3-phenylpropionitrile + H2O | 59% activity compared to glutaronitrile | uncultured bacterium | ? | - |
? | |
| acetonitrile + H2O | 5% activity compared to glutaronitrile | uncultured bacterium | ? | - |
? | |
| adiponitrile + H2O | 91% activity compared to glutaronitrile | uncultured bacterium | adipate + NH3 | - |
? | |
| butyronitrile + H2O | 6% activity compared to glutaronitrile | uncultured bacterium | ? | - |
? | |
| cinnamonitrile + H2O | 28% activity compared to glutaronitrile | uncultured bacterium | cinnamate + NH3 | - |
? | |
| crotonitrile + H2O | 7% activity compared to glutaronitrile | uncultured bacterium | ? | - |
? | |
| glutaronitrile + H2O | 100% activity | uncultured bacterium | glutarate + NH3 | - |
? | |
| additional information | Nit1 does not hydrolyze malononitrile, mandelonitrile, 4-cyanobenzoic acid, 3-ethoxybenzonitrile, 4-hydroxybenzonitrile, alpha-methylbenzyl cyanide, acrylonitrile, 4-tolunitrile, and benzonitrile | uncultured bacterium | ? | - |
? | |
| additional information | Nit1 is an aliphatic nitrilase favoring dinitriles over mononitriles | uncultured bacterium | ? | - |
? | |
| propionitrile + H2O | 5% activity compared to glutaronitrile | uncultured bacterium | ? | - |
? | |
| succinonitrile + H2O | 76% activity compared to glutaronitrile | uncultured bacterium | succinate + NH3 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 38000, SDS-PAGE | uncultured bacterium |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NIT1 | - |
uncultured bacterium |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
using the substrate glutaronitrile | uncultured bacterium |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | 60 | up to 40°C Nit1 is stable for 1 h, while the loss of activity is about 50% after 30 min at 50°C, the enzyme is completely inactivated after 10 min incubation at 60°C | uncultured bacterium |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
using the substrate glutaronitrile | uncultured bacterium |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 9 | Nit1 tolerates a pH range between 5 and 9 with an activity of more than 50% | uncultured bacterium |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.483 | - |
glutaronitrile | at 30°C in 50 mM sodium phosphate buffer (pH 7.5) and 1 mM dithiothreitol | uncultured bacterium | |
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