Application | Comment | Organism |
---|---|---|
environmental protection | degradation of nitrile waste | Rhodococcus sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetamide | 10 mM, product inhibition, reduction of acrylonitrile degrading activity from 1.2 micromol/min/mg to 1.0 micromol/min/mg | Rhodococcus sp. | |
CuCl2 | 1 mM, strong inhibition of EDTA-dialysed enzyme, 19% remaining activity | Rhodococcus sp. | |
ethanolamine | markedly lower activity | Rhodococcus sp. | |
Fe2+ | 1 mM, inhibition of EDTA-dialysed enzyme, 38% remaining activity | Rhodococcus sp. | |
propioamide | 10 mM, product inhibition, reduction of acrylonitrile degrading activity from 1.2 micromol/min/mg to 0.91 micromol/min/mg | Rhodococcus sp. | |
Zn2+ | 1 mM, inhibition of EDTA-dialysed enzyme, 56% remaining activity | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
Propionitrile | +/- 0.1, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. | |
1.9 | - |
Butyronitrile | +/- 0.2, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. | |
5.4 | - |
acetonitrile | +/- 0.3, 10 mM HEPES, pH 7.5, 37°C, Michaelis-Menten behavior at acetonitrile concentrations from 0.1 mM to 50 mM | Rhodococcus sp. | |
6.4 | - |
acrylonitrile | +/- 0.3, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | component of enzyme, one ion per holoenzyme, ICP-MS | Rhodococcus sp. | |
Co2+ | increase of activity of EDTA-dialysed enzyme, 120% activity | Rhodococcus sp. | |
Cu2+ | component of enzyme, two ions per holoenzyme, ICP-MS | Rhodococcus sp. | |
Mg2+ | increase of activity of EDTA-dialysed enzyme, 130% activity | Rhodococcus sp. | |
additional information | metal content is unusual for NHases | Rhodococcus sp. | |
Zn2+ | component of enzyme, one ion per holoenzyme, ICP-MS | Rhodococcus sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
55746 | - |
alpha,beta2, 1 * 55746 + 2 * 63001, 32% amino acid sequence identity of subunits, MALDI-TOF MS and HPLC | Rhodococcus sp. |
63001 | - |
alpha,beta2, 1 * 55746 + 2 * 63001, 32% amino acid sequence identity of subunits, MALDI-TOF MS and HPLC | Rhodococcus sp. |
364000 | - |
proposal for holoenzyme subunit composition: alpha2 beta4, based on calibrated gel filtration and absorption spectra | Rhodococcus sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | - |
- |
- |
Rhodococcus sp. RHA1 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
anion exchange and hydrophobic interaction chromatography, yield of purified protein: 1.9% (specific activity 5.9 U/mg), cytosolic proteom of cells grown on 0.1 M acetonitrile and acetic acid/ammonia are compared via quantitative 2D gel electrophoresis, spot identification by MS, no significant amino acid sequence similarity with other NHases | Rhodococcus sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.6 | - |
+/- 0.2, acrylonitrile, 50 mM, coupled assay with amidase | Rhodococcus sp. |
2.8 | - |
+/- 0.2, acrylonitrile, 50 mM, HPLC-based assay | Rhodococcus sp. |
5.7 | - |
+/- 0.2, acetonitrile, 50 mM, HPLC-based assay | Rhodococcus sp. |
5.9 | - |
+/- 0.4, acetonitrile, 50 mM, coupled assay with amidase | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetonitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. | acetamide | - |
r | |
acetonitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. RHA1 | acetamide | - |
r | |
acrylonitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. | acrylamide | - |
r | |
acrylonitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. RHA1 | acrylamide | - |
r | |
butyronitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. | butyramide | - |
r | |
butyronitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. RHA1 | butyramide | - |
r | |
additional information | no detectably transformation with 2-methyl-2-butenenitrile, benzonitrile and phenylacetonitrile | Rhodococcus sp. | ? | - |
? | |
additional information | no detectably transformation with 2-methyl-2-butenenitrile, benzonitrile and phenylacetonitrile | Rhodococcus sp. RHA1 | ? | - |
? | |
propionitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. | propionamide | - |
r | |
propionitrile + H2O | substrate specificity: acetonitrile ~ propionitrile > acrylonitrile >> butyronitrile | Rhodococcus sp. RHA1 | propionamide | - |
r |
Subunits | Comment | Organism |
---|---|---|
heterotrimer | alpha,beta2, 1 * 55746 + 2 * 63001, 32% amino acid sequence identity of subunits, MALDI-TOF MS and HPLC | Rhodococcus sp. |
More | sequence similarity and size of subunits is unusual for NHases | Rhodococcus sp. |
Synonyms | Comment | Organism |
---|---|---|
ANHase | acetonitrile hydratase | Rhodococcus sp. |
NHase | - |
Rhodococcus sp. |
nitrile hydratase | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.99 | - |
Butyronitrile | +/- 0.04, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. | |
2 | 8 | acrylonitrile | +/- 0.5, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. | |
2.6 | - |
Propionitrile | +/- 0.2, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. | |
41 | - |
acetonitrile | +/- 0.9, 10 mM HEPES, pH 7.5, 37°C | Rhodococcus sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Rhodococcus sp. |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5.5 | 10 | activity drops sharply below pH 5.5 and above pH 10.0 | Rhodococcus sp. |