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Literature summary for 4.2.1.84 extracted from

  • Sari, M.A.; Jaouen, M.; Saroja, N.R.; Artaud, I.
    Influence of cobalt substitution on the activity of iron-type nitrile hydratase: are cobalt type nitrile hydratases regulated by carbon monoxide? (2007), J. Inorg. Biochem., 101, 614-622.
    View publication on PubMed
Search for more literature for 4.2.1.84

Cloned(Commentary)

Cloned (Comment) Organism
nitrile hydratase from Comamonas testosteroni is expressed in Escherichia coli, strain TG1, overexpressed chaperonin system Comamonas testosteroni

Protein Variants

Protein Variants Comment Organism
additional information Fe of an Fe-dependent recombinant nitrile hydratase (wild-type, NilFe) is replaced by Co generating an Co-substituted enzyme (NilCo) Comamonas testosteroni

Inhibitors

Inhibitors Comment Organism Structure
2-Nitrophenol 10 microM, 40-50% residual activity of NilFe, 90% residual activity of NilCo Comamonas testosteroni
3-Nitrophenol 10 microM, 40-50% residual activity of NilFe, fully active NilCo Comamonas testosteroni
4-nitrophenol 10 microM, 40-50% residual activity of NilFe, 80% residual activity of NilCo Comamonas testosteroni
7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole
-
 Comamonas testosteroni
acrylonitrile
-
 Comamonas testosteroni
Butyric acid
-
 Comamonas testosteroni
CO inhibition of NilCo, reversible by photoactivation, no inhibition of NilFe Comamonas testosteroni
KCN
-
 Comamonas testosteroni
NaNO2 NilFe is unaffected by NaNO2 Comamonas testosteroni
NO inhibition of NilCo and NilFe, reversible by photoactivation with 150 W white light Comamonas testosteroni
tertiary-butyl isonitrile NilFe is unaffected by tertiary-butyl isonitrile Comamonas testosteroni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
 (25R)-3beta-hydroxycholest-5-en-27-oate +/- 0.04, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilFe Comamonas testosteroni
0.19
-
 (25R)-3beta-hydroxycholest-5-en-27-oate +/- 0.06, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilCo Comamonas testosteroni

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
22700
-
 alpha2beta2, 2 * 22800 + 2 * 22700 Comamonas testosteroni
22800
-
 alpha2beta2, 2 * 22800 + 2 * 22700 Comamonas testosteroni
90000
-
 gel filtration Comamonas testosteroni

Organism

Organism UniProt Comment Textmining
Comamonas testosteroni
-
 Fe-dependent, wild-type nitrile hydratase is compared with artificial, Co-dependent nitrile hydratase
-

Purification (Commentary)

Purification (Comment) Organism
CoCl2, an overexpressed chaperonin system and butyric acid is necessary for fully soluble and active NilCo protein, 95% purity on 12% SDS-PAGE Comamonas testosteroni

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 vmax: 28 +/- 5 nmol/min/mg, NilCo Comamonas testosteroni
additional information
-
 vmax: 30 +/- 5 nmol/min/mg, NilFe Comamonas testosteroni
0.018
-
 0.5 mM NaNO2, NilFe Comamonas testosteroni
15
-
 +/- 5, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilCo Comamonas testosteroni
15
-
 +/- 5, substrate: 2-nitro-5-thiocyanato-benzoic acid, NilFe Comamonas testosteroni
100
-
 +/- 15, substrate: 5-cyanovaleric acid, NilFe Comamonas testosteroni
110
-
 +/- 10, substrate: 5-cyanovaleric acid, NilCo Comamonas testosteroni

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(25R)-3beta-hydroxycholest-5-en-27-oate
-
 Comamonas testosteroni ?
-
 ?
5-cyanovaleric acid + H2O NilFe and NilCo Comamonas testosteroni 6-amino-6-oxohexanoic acid
-
 r

Subunits

Subunits Comment Organism
tetramer alpha2beta2, 2 * 22800 + 2 * 22700 Comamonas testosteroni

Synonyms

Synonyms Comment Organism
NHase
-
 Comamonas testosteroni
NilCo
-
 Comamonas testosteroni
NilFe
-
 Comamonas testosteroni
nitrile hydratase
-
 Comamonas testosteroni

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.9 7.2 NilFe, NilCo Comamonas testosteroni

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.002
-
 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole +/- 0.0005, NilFe Comamonas testosteroni
0.002
-
 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole +/- 0.0008, NilCo Comamonas testosteroni
0.0025
-
 KCN +/- 0.001, NilCo Comamonas testosteroni
0.003
-
 NaNO2 +/- 0.0005, NilCo Comamonas testosteroni
0.003
-
 KCN +/- 0.0015, NilFe Comamonas testosteroni
0.004
-
 tertiary-butyl isonitrile +/- 0.001, NilCo Comamonas testosteroni
0.005
-
 acrylonitrile +/- 0.0005, NilCo Comamonas testosteroni
0.007
-
 acrylonitrile +/- 0.001, NilFe Comamonas testosteroni

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.003
-
 +/- 0.001, NilCo Comamonas testosteroni KCN
0.003
-
 +/- 0.001, NilCo Comamonas testosteroni 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole
0.004
-
 +/- 0.001, NilFe Comamonas testosteroni 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole
0.005
-
 +/- 0.001, NilFe Comamonas testosteroni KCN
0.005
-
 +/- 0.001, NilFe Comamonas testosteroni 2-Nitrophenol
0.005
-
 +/- 0.003, NilCo Comamonas testosteroni acrylonitrile
0.007
-
 +/- 0.001, NilCo Comamonas testosteroni tertiary-butyl isonitrile
0.008
-
 +/- 0.001, NilCo Comamonas testosteroni NaNO2
0.0086
-
 pH 7.9, NilCo Comamonas testosteroni 4-nitrophenol
0.009
-
 +/- 0.002, NilFe Comamonas testosteroni acrylonitrile
0.01
-
 +/- 0.002, NilFe Comamonas testosteroni 4-nitrophenol
0.01
-
 +/- 0.002, NilFe Comamonas testosteroni 3-Nitrophenol
0.1
-
 value above, NilFe Comamonas testosteroni tertiary-butyl isonitrile
0.3
-
 +/- 0.1, NilCo Comamonas testosteroni Butyric acid
0.4
-
 +/- 0.1, NilFe Comamonas testosteroni Butyric acid
0.5
-
 value above, NilFe Comamonas testosteroni NaNO2