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Literature summary for 4.2.1.84 extracted from

  • Takarada, H.; Kawano, Y.; Hashimoto, K.; Nakayama, H.; Ueda, S.; Yohda, M.; Kamiya, N.; Dohmae, N.; Maeda, M.; Odaka, M.
    Mutational study on alphaGln90 of Fe-type nitrile hydratase from Rhodococcus sp. N771 (2006), Biosci. Biotechnol. Biochem., 70, 881-889.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
x-ray diffraction, special focus on iron center vicinity and hydrogen bond network, similar structures of wild type and alphaQ90N mutant enzyme in nitrosylated state, resolution: 1.43 A Rhodococcus sp.

Protein Variants

Protein Variants Comment Organism
Q90E alpha subunit Rhodococcus sp.
Q90N alpha subunit Rhodococcus sp.
R56K beta subunit Rhodococcus sp.

General Stability

General Stability Organism
purified mutant NHases are stored in the dark without n-butyric acid, before use, NHases are activated by light irradiation. Rhodococcus sp.

Inhibitors

Inhibitors Comment Organism Structure
n-butyric acid
-
Rhodococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.28
-
Methacrylonitrile mutant alphaQ90E, vmax: 280 micromol/min/mg Rhodococcus sp.
0.68
-
Methacrylonitrile wild type, vmax: 1200 micromol/min/mg Rhodococcus sp.
1.9
-
Methacrylonitrile mutant betaR56K, vmax: 2.5 micromol/min/mg Rhodococcus sp.
2.6
-
Methacrylonitrile mutant alphaQ90N, vmax: 57 micromol/min/mg Rhodococcus sp.

Organism

Organism UniProt Comment Textmining
Rhodococcus sp.
-
-
-
Rhodococcus sp. N771
-
-
-

Purification (Commentary)

Purification (Comment) Organism
3 successive column chromatographies in the presence of 40 mM n-butyric acid (only wild type), alphaQ90N mutein is purified in nitrosylated state in the dark Rhodococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methacrylonitrile + H2O
-
Rhodococcus sp. methacrylamide
-
?
methacrylonitrile + H2O
-
Rhodococcus sp. N771 methacrylamide
-
?

Synonyms

Synonyms Comment Organism
NHase
-
Rhodococcus sp.
nitrile hydratase
-
Rhodococcus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
120
-
Methacrylonitrile mutant betaR56K Rhodococcus sp.
2600
-
Methacrylonitrile mutant alphaQ90N Rhodococcus sp.
13000
-
Methacrylonitrile mutant alphaQ90E Rhodococcus sp.
55000
-
Methacrylonitrile wild type Rhodococcus sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.6
-
Butyric acid +/- 0.2, substrate: methacrylonitrile, mutant betaR56K Rhodococcus sp.
5.1
-
Butyric acid +/- 0.3, substrate: methacrylonitrile, mutant alphaQ90E Rhodococcus sp.
6.2
-
Butyric acid +/- 0.9, substrate: methacrylonitrile, wild type Rhodococcus sp.
14
-
Butyric acid +/- 2.7, substrate: methacrylonitrile, mutant alphaQ90N Rhodococcus sp.