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Literature summary for 3.5.5.7 extracted from

  • Fan, H.; Chen, L.; Sun, H.; Wang, H.; Ren, Y.; Wei, D.
    A novel nitrilase from Ralstonia eutropha H16 and its application to nicotinic acid production (2017), Bioprocess Biosyst. Eng., 40, 1271-1281 .
    View publication on PubMed

Application

Application Comment Organism
industry under optimized conditions, using the fed-batch reaction mode, total of 1050 mM 3-cyanopyridine is hydrolyzed completely in 20.8 h with eight substrate feedings, yielding 129.2 g/l production of nicotinic acid and thus showing a potential for industrial application Cupriavidus necator
synthesis under optimized conditions, using the fed-batch reaction mode, total of 1050 mM 3-cyanopyridine is hydrolyzed completely in 20.8 h with eight substrate feedings, yielding 129.2 g/l production of nicotinic acid and thus showing a potential for industrial application Cupriavidus necator

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli BL21(DE3) Cupriavidus necator

Inhibitors

Inhibitors Comment Organism Structure
acetone 25% (v/v), about 40% inhibition Cupriavidus necator
Ag+ 5 mM, 94% inhibition Cupriavidus necator
Ca2+ 5 mM, 97% inhibition Cupriavidus necator
Co2+ 5 mM, 15% inhibition Cupriavidus necator
Cu2+ 5 mM, 70% inhibition Cupriavidus necator
dimethyl sulfoxide 15% (v/v),about 35% inhibition Cupriavidus necator
ethanol 25% (v/v), about 20% inhibition Cupriavidus necator
Fe2+ 5 mM, 51% inhibition Cupriavidus necator
Isopropanol 25% (v/v), about 10% inhibition Cupriavidus necator
Li+ 5 mM, 37% inhibition Cupriavidus necator
methanol 25% (v/v), about 20% inhibition Cupriavidus necator
Mg2+ 5 mM, 31% inhibition Cupriavidus necator
Mn2+ 5 mM, 61% inhibition Cupriavidus necator
Ni2+ 5 mM, 47% inhibition Cupriavidus necator
tetrahydrofuran 10% (v/v), about 95% inhibition Cupriavidus necator
Zn2+ 5 mM, 47% inhibition Cupriavidus necator

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.4
-
3-Cyanopyridine pH 7.0, 30°C Cupriavidus necator

Organism

Organism UniProt Comment Textmining
Cupriavidus necator
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-cyanopyridine + 2 H2O activity is 3.35fold higher compared to activity with 3-cyanopyridine Cupriavidus necator pyridine 2-carboxylic acid + NH3
-
?
2-methylglutaronitrile + 2 H2O activity is 10% compared to activity with 3-cyanopyridine Cupriavidus necator 4-cyanopentanoic acid + NH3
-
?
2-phenylbutyronitrile + 2 H2O activity is 11% compared to activity with 3-cyanopyridine Cupriavidus necator 2-phenylpropionic acid + NH3
-
?
3-cyanopyridine + 2 H2O
-
Cupriavidus necator pyridine 3-carboxylic acid + NH3
-
?
3-hydroxyglutaronitrile + 4 H2O activity is 38% compared to activity with 3-cyanopyridine Cupriavidus necator 3-hydroxyglutaric acid + 2 NH3
-
?
3-hydroxypropionitrile + 2 H2O activity is 1.5fold higher compared to activity with 3-cyanopyridine Cupriavidus necator 3-hydroxypropionitrile + NH3
-
?
3-phenylpropionitrile + 2 H2O activity is 61% compared to activity with 3-cyanopyridine Cupriavidus necator 3-phenylpropionic acid + NH3
-
?
4-chlorobutyronitrile + 2 H2O activity is 1.49fold higher compared to activity with 3-cyanopyridine Cupriavidus necator 4-chlorobutyric acid + NH3
-
?
4-cyanopyridine + 2 H2O activity is 3fold higher compared to activity with 3-cyanopyridine Cupriavidus necator pyridine 4-carboxylic acid + NH3
-
?
adiponitrile + 4 H2O activity is 1.14fold higher compared to activity with 3-cyanopyridine Cupriavidus necator adipic acid + 2 NH3
-
?
cinnamonitrile + 2 H2O activity is 8fold higher compared to activity with 3-cyanopyridine Cupriavidus necator cinnamic acid + NH3
-
?
fumaronitrile + 4 H2O activity is 36fold higher compared to activity with 3-cyanopyridine Cupriavidus necator fumaric acid + 2 NH3
-
?
glycolonitrile + 2 H2O activity is 15.9fold higher compared to activity with 3-cyanopyridine Cupriavidus necator glycolic acid + NH3
-
?
iminodiacetonitrile + 4 H2O activity is 39% compared to activity with 3-cyanopyridine Cupriavidus necator iminodiacetic acid + 2 NH3
-
?
indole-3-acetonitrile + 2 H2O activity is 48% compared to activity with 3-cyanopyridine Cupriavidus necator indol-3-acetic acid + NH3
-
?
additional information the enzyme shows a broad hydrolytic activity toward aliphatic and heterocyclic nitriles and shows high tolerance of 3-cyanopyridine. No activity with: benzonitrile, phenylacetonitrile, 4-chlorobenzyl cyanide, mandelonitrile, 2-chloromandelonitrile, alpha-methylphenylacetonitrile, 1,2-phenylenediacetonitrile, dodecanenitrile Cupriavidus necator ?
-
-
pimelonitrile + 4 H2O activity is 85% compared to activity with 3-cyanopyridine Cupriavidus necator pimelic acid + 2 NH3
-
?
sebaconitrile + 4 H2O activity is 1.66fold higher compared to activity with 3-cyanopyridine Cupriavidus necator sebaconic acid + 2 NH3
-
?
succinonitrile + 4 H2O activity is 1.97fold higher compared to activity with 3-cyanopyridine Cupriavidus necator succinic acid + 2 NH3
-
?
valeronitrile + 2 H2O activity is 99% compared to activity with 3-cyanopyridine Cupriavidus necator valeric acid + NH3
-
?

Synonyms

Synonyms Comment Organism
REH16
-
Cupriavidus necator

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Cupriavidus necator

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 50 20°C: about 60% of maximal activity, 50°C: about 85% of maximal activity Cupriavidus necator

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25
-
120 h, 5% loss of activity Cupriavidus necator

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.6
-
-
Cupriavidus necator

pH Range

pH Minimum pH Maximum Comment Organism
5 8.5 pH 5.0: about 40% of maximal activity, pH 8.5: about 55% of maximal activity Cupriavidus necator