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Literature summary for 3.5.5.5 extracted from
- A novel nitrilase, arylacetonitrilase, of Alcaligenes faecalis JM3. Purification and characterization (1990), Eur. J. Biochem., 194, 765-772.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | activation | Alcaligenes faecalis |  |
| dithiothreitol | activation | Alcaligenes faecalis | |
| Sodium hydrosulfide | activation | Alcaligenes faecalis | |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2,2'-dipyridyl | weak | Alcaligenes faecalis | |
| 3-Methyl-2-benzothiazolinone hydrazone | weak | Alcaligenes faecalis | |
| 8-hydroxyquinoline | weak | Alcaligenes faecalis | |
| Ag2+ | strong | Alcaligenes faecalis | |
| ascorbic acid | - |
Alcaligenes faecalis | |
| Cd2+ | - |
Alcaligenes faecalis | |
| Co2+ | - |
Alcaligenes faecalis | |
| Cu2+ | strong | Alcaligenes faecalis | |
| D-cycloserine | weak | Alcaligenes faecalis | |
| diethyldithiocarbamate | - |
Alcaligenes faecalis | |
| Disodium 4,5-dihydroxy-m-benzene-disulfonate | weak | Alcaligenes faecalis | |
| DTNB | strong | Alcaligenes faecalis | |
| H2O2 | - |
Alcaligenes faecalis | |
| Hg2+ | strong | Alcaligenes faecalis | |
| hydroxylamine | weak | Alcaligenes faecalis | |
| iodoacetate | weak | Alcaligenes faecalis | |
| KCN | weak | Alcaligenes faecalis | |
| L-cysteine | - |
Alcaligenes faecalis | |
| additional information | little or no inhibition by Li+, Na+, Mg2+, Ca2+, Ba2+, Mn2+, Fe2+, Pb2+, Fe3+, Al3+, NaN3, NaF, cysteamine, aminoguanidine, L/D-penicillamine, 1,10-phenanthroline or EDTA | Alcaligenes faecalis | |
| N-ethylmaleimide | - |
Alcaligenes faecalis | |
| Ni2+ | - |
Alcaligenes faecalis | |
| p-chloromercuribenzoate | strong | Alcaligenes faecalis | |
| phenylhydrazine | - |
Alcaligenes faecalis | |
| Semicarbazide | weak | Alcaligenes faecalis | |
| Sn2+ | - |
Alcaligenes faecalis | |
| Urea | weak | Alcaligenes faecalis | |
| Zn2+ | - |
Alcaligenes faecalis | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.009 | - |
3-Indoleacetonitrile | - |
Alcaligenes faecalis | |
| 0.01 | - |
benzylcyanide | - |
Alcaligenes faecalis | |
| 0.015 | - |
2-Chlorobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.016 | - |
3-Thiopheneacetonitrile | - |
Alcaligenes faecalis | |
| 0.022 | - |
4-Aminobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.023 | - |
3-tolylacetonitrile | - |
Alcaligenes faecalis | |
| 0.065 | - |
3-Chlorobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.091 | - |
2-Thiopheneacetonitrile | - |
Alcaligenes faecalis | |
| 0.105 | - |
4-Nitrobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.123 | - |
4-tolylacetonitrile | - |
Alcaligenes faecalis | |
| 0.13 | - |
4-Fluorobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.333 | - |
4-Chlorobenzylcyanide | - |
Alcaligenes faecalis | |
| 0.66 | - |
acrylonitrile | - |
Alcaligenes faecalis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| additional information | - |
amino acid composition | Alcaligenes faecalis |
| 44000 | - |
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition | Alcaligenes faecalis |
| 47770 | - |
6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition | Alcaligenes faecalis |
| 260000 | - |
Alcaligenes faecalis, gel filtration on Sepharose 6B | Alcaligenes faecalis |
| 275000 | - |
Alcaligenes faecalis, gel permeation HPLC | Alcaligenes faecalis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Arylacetonitrile + H2O | Alcaligenes faecalis | pathway in nitrile catabolism, inducible enzyme | ? | - |
? | |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| Methanol | stable in the presence of 10% w/v methanol | Alcaligenes faecalis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Alcaligenes faecalis | - |
JM3 | - |
| Alcaligenes faecalis JM3 | - |
JM3 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Alcaligenes faecalis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 144 | - |
- |
Alcaligenes faecalis |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 5 months | Alcaligenes faecalis |
| 4°C, in 10 mM potassium phosphate buffer, pH 7, 10 mM DTT and 50% glycerol, at least 2 months, without glycerol 60% loss of activity within 1 week | Alcaligenes faecalis |
| 4°C, in the presence of 60% saturated ammonium sulfate, 2 M NaCl or 30% w/v propanediol, only about 5% loss of activity within 1 week | Alcaligenes faecalis |
| 5°C, in 0.02 M citrate buffer, pH 7-8, 10 days, about 50% loss of activity in 0.02 M Tris-HCl buffer, pH 9.3 within 10 days | Alcaligenes faecalis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1-cyclohexenylacetonitrile + H2O | at 3.3% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 1-cyclohexenylacetate + NH3 | - |
? | |
| 2,4-dichlorobenzylcyanide + H2O | at 6.5% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 2,4-dichlorophenylacetic acid + NH3 | - |
? | |
| 2-chlorobenzylcyanide + H2O | at 6% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 2-chlorophenylacetic acid + NH3 | - |
? | |
| 2-methoxybenzylcyanide + H2O | at 2% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 2-methoxyphenylacetic acid + NH3 | - |
? | |
| 2-thiopheneacetonitrile + H2O | ir, at 90% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 2-thiopheneacetic acid + NH3 | not via thiopheneacetamide | ? | |
| 3,4-(methylenedioxy)phenylacetonitrile + H2O | at 65% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3,4-(methylendioxy)phenylacetic acid + NH3 | - |
? | |
| 3,4-dichlorobenzylcyanide + H2O | at 14% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3,4-dichlorophenylacetic acid + NH3 | - |
? | |
| 3-chlorobenzylcyanide + H2O | at 43% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3-chlorophenylacetic acid + NH3 | - |
? | |
| 3-indoleacetonitrile + H2O | at 9% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3-indoleacetic acid + NH3 | - |
? | |
| 3-indoleacetonitrile + H2O | at 9% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis JM3 | 3-indoleacetic acid + NH3 | - |
? | |
| 3-phenylpropionitrile + H2O | at 26% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3-phenylpropionic acid + NH3 | - |
? | |
| 3-pyridineacetonitrile + H2O | at 77% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | ? | - |
? | |
| 3-thiopheneacetonitrile + H2O | at 24% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 3-thiopheneacetic acid + NH3 | - |
? | |
| 4-aminobenzylcyanide + H2O | at 44% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 4-aminophenylacetic acid + NH3 | - |
? | |
| 4-bromobenzylcyanide + H2O | at 26% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | ? | - |
? | |
| 4-bromobenzylcyanide + H2O | at 26% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis JM3 | ? | - |
? | |
| 4-chlorobenzylcyanide + H2O | i.e. p-chlorophenylacetonitrile, best substrate | Alcaligenes faecalis | 4-chlorophenylacetic acid + NH3 | - |
? | |
| 4-chlorobenzylcyanide + H2O | i.e. p-chlorophenylacetonitrile, best substrate | Alcaligenes faecalis JM3 | 4-chlorophenylacetic acid + NH3 | - |
? | |
| 4-fluorobenzylcyanide + H2O | i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 4-fluorophenylacetic acid + NH3 | - |
? | |
| 4-fluorobenzylcyanide + H2O | i.e. p-fluorophenylacetonitrile, at 96% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis JM3 | 4-fluorophenylacetic acid + NH3 | - |
? | |
| 4-hydroxybenzylcyanide + H2O | at 29% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 4-hydroxyphenylacetic acid + NH3 | - |
? | |
| 4-methoxybenzylcyanide + H2O | at 62% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 4-methoxyphenylacetic acid + NH3 | - |
? | |
| 4-nitrobenzylcyanide + H2O | at 45% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | 4-nitrophenylacetate + NH3 | - |
? | |
| acrylonitrile + H2O | at 7% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | propenoic acid + NH3 | - |
? | |
| Arylacetonitrile + H2O | pathway in nitrile catabolism, inducible enzyme | Alcaligenes faecalis | ? | - |
? | |
| benzylcyanide + H2O | at 53% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | phenylacetic acid + NH3 | - |
? | |
| m-tolylacetonitrile + H2O | at 32% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | m-tolylacetic acid + NH3 | - |
? | |
| m-xylenedicyanide + H2O | at 2.7% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | ? | - |
? | |
| mandelonitrile + 2 H2O | at 8% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | mandelic acid + NH3 | - |
? | |
| additional information | very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile | Alcaligenes faecalis | ? | - |
? | |
| additional information | very poor substrates (less than 1.5% the rate of 4-chlorobenzylcyanide hydrolysis) are 1,4-dichlorobenzylcyanide, 4-nitrobenzylcyanide, 2-pyridineacetonitrile, 2-phenylpropionitrile, 1-naphthylacetonitrile, diphenylacetonitrile, o-xylenedicyanide, acetonitrile (i.e. methylcyanide). No substrates are 2,6-dichlorobenzylcyanide, allylcyanide, propionitrile, aminoacetonitrile, iminodiacetonitrile, methacrylonitrile, crotonitrile, benzonitrile, 2- or 3-cyanopyridine, cyanopyrazine, 2-thiophenecarbonitrile or 2-furaneacrylonitrile | Alcaligenes faecalis JM3 | ? | - |
? | |
| o-tolylacetonitrile + H2O | at 4% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | o-tolylacetic acid + NH3 | - |
? | |
| p-tolylacetonitrile + H2O | at 72% the rate of 4-chlorobenzylcyanide hydrolysis | Alcaligenes faecalis | p-tolylacetic acid + NH3 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| hexamer | 6 * 44000, Alcaligenes faecalis, SDS-PAGE, 6 * 47770, Alcaligenes faecalis, minimal value derived from amino acid composition | Alcaligenes faecalis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | 30 | assay at | Alcaligenes faecalis |
| 45 | - |
- |
Alcaligenes faecalis |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
at 50°C and above, rapid loss of activity | Alcaligenes faecalis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
and below, 30 min, stable in 10 mM potassium phosphate buffer, pH 7.5 | Alcaligenes faecalis |
| 35 | - |
30 min, in 10 mM potassium phosphate buffer, pH 7.5, 25% loss of activity | Alcaligenes faecalis |
| 40 | - |
30 min, in 10 mM potassium phosphate buffer, pH 7.5, inactivation | Alcaligenes faecalis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
2-thiopheneacetonitrile as substrate | Alcaligenes faecalis |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6.4 | 9.4 | about half-maximal activity at pH 6.4 and 9.4, 2-thiopheneacetonitrile as substrate | Alcaligenes faecalis |
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