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Literature summary for 3.5.5.1 extracted from

  • Zhu, X.; Gong, J.; Li, H.; Lu, Z.; Zhou, Z.; Shi, J.; Xu, Z.
    Characterization and functional cloning of an aromatic nitrilase from Pseudomonas putida CGMCC3830 with high conversion efficiency toward cyanopyridine (2013), J. Mol. Catal. B, 97, 175-183.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta-gami (DE3) cells Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
Ag+ complete inhibition at 1 mM Pseudomonas putida
ascorbic acid 68% residual activity at 1 mM Pseudomonas putida
Co2+ 44.9% residual activity at 1 mM Pseudomonas putida
Cu2+ 19.7% residual activity at 1 mM Pseudomonas putida
EDTA 93.2% residual activity at 1 mM Pseudomonas putida
Fe2+ 75.4% residual activity at 1 mM Pseudomonas putida
Fe3+ 53.3% residual activity at 1 mM Pseudomonas putida
Hg+ complete inhibition at 1 mM Pseudomonas putida
iodoacetamide 56.6% residual activity at 1 mM Pseudomonas putida
Mn2+ 67.4% residual activity at 1 mM Pseudomonas putida
additional information not inhibited by L-glutathione, L-cysteine, dithiothreitol, and 8-hydroxyquinoline Pseudomonas putida
N-ethylmaleimide 57.3% residual activity at 1 mM Pseudomonas putida
Ni2+ 21.6% residual activity at 1 mM Pseudomonas putida
Zn2+ 57% residual activity at 1 mM Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
27.9
-
3-Cyanopyridine at pH 7.5 and 30°C Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
41000
-
8 * 41000, SDS-PAGE Pseudomonas putida
310000
-
gel filtration Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-cyanopyridine + 2 H2O Pseudomonas putida
-
nicotinic acid + NH3
-
?
3-cyanopyridine + 2 H2O Pseudomonas putida CGMCC3830
-
nicotinic acid + NH3
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-
Pseudomonas putida CGMCC3830
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, butyl-Sepharose column chromatography, Q-Sepharose column chromatography, and Superdex 200 gel filtration Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-cyanopyridine + H2O 75.9% activity compared to 3-cyanopyridine Pseudomonas putida ?
-
?
2-cyanopyridine + 2 H2O 1.5% activity compared to 3-cyanopyridine Pseudomonas putida pyridine 2-carboxylic acid + NH3
-
?
3-cyanopyridine + 2 H2O
-
Pseudomonas putida nicotinic acid + NH3
-
?
3-cyanopyridine + 2 H2O 100% activity Pseudomonas putida nicotinic acid + NH3
-
?
3-cyanopyridine + 2 H2O
-
Pseudomonas putida CGMCC3830 nicotinic acid + NH3
-
?
4-cyanopyridine + 2 H2O 79.5% activity compared to 3-cyanopyridine Pseudomonas putida pyridine 4-carboxylic acid + NH3
-
?
4-methoxyphenylacetonitrile + H2O 0.6% activity compared to 3-cyanopyridine Pseudomonas putida 4-methoxyphenylacetic acid + NH3
-
?
acetonitrile + H2O 1.0% activity compared to 3-cyanopyridine Pseudomonas putida acetic acid + NH3
-
?
acrylonitrile + H2O 17.2% activity compared to 3-cyanopyridine Pseudomonas putida acrylic acid + NH3
-
?
acrylonitrile + H2O 17.2% activity compared to 3-cyanopyridine Pseudomonas putida CGMCC3830 acrylic acid + NH3
-
?
aminoacetonitrile + H2O 9.0% activity compared to 3-cyanopyridine Pseudomonas putida aminoacetic acid + NH3
-
?
benzonitrile + 2 H2O 70.4% activity compared to 3-cyanopyridine Pseudomonas putida benzoic acid + NH3
-
?
benzonitrile + 2 H2O 70.4% activity compared to 3-cyanopyridine Pseudomonas putida CGMCC3830 benzoic acid + NH3
-
?
beta-aminopropionitrile + H2O 8.6% activity compared to 3-cyanopyridine Pseudomonas putida beta-aminopropionic acid + NH3
-
?
iminodiacetonitrile + 2 H2O 27% activity compared to 3-cyanopyridine Pseudomonas putida iminodiacetic acid + 2 NH3
-
?
iminodiacetonitrile + 2 H2O 27% activity compared to 3-cyanopyridine Pseudomonas putida CGMCC3830 iminodiacetic acid + 2 NH3
-
?
additional information no activity with isopentanenitrile, adiponitrile, 2-chloro-3-cyanopyridine, and mandelonitrile Pseudomonas putida ?
-
?
additional information no activity with isopentanenitrile, adiponitrile, 2-chloro-3-cyanopyridine, and mandelonitrile Pseudomonas putida CGMCC3830 ?
-
?
pentanenitrile + H2O 1.6% activity compared to 3-cyanopyridine Pseudomonas putida ?
-
?
pyrazinecarbonitrile + H2O 32.7% activity compared to 3-cyanopyridine Pseudomonas putida ?
-
?
succinonitrile + H2O 21.74% activity compared to 3-cyanopyridine Pseudomonas putida succinic acid + NH3
-
?

Subunits

Subunits Comment Organism
homooctamer 8 * 41000, SDS-PAGE Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Pseudomonas putida

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 more than 70% activity between 30 and 50°C. A reaction temperature of 55°C leads to almost totally inactivation of the enzyme Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Pseudomonas putida

pH Range

pH Minimum pH Maximum Comment Organism
7 8 more than 80% activity between pH 7.0 and 8.0 Pseudomonas putida

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas putida isoelectric focusing
-
5.2