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Literature summary extracted from

  • Cheng, Z.; Jiang, S.; Zhou, Z.
    Substrate access tunnel engineering for improving the catalytic activity of a thermophilic nitrile hydratase toward pyridine and pyrazine nitriles (2021), Biochem. Biophys. Res. Commun., 575, 8-13.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.84 L48H mutation of residue located at the substrate access tunnel of subunit NthB, specific activities towards 3-cyanopyridine, 2-cyanopyridine and cyanopyrazine are 2.4fold, 2.8fold and 3.1fold higher tha that of wild-type. Introduced H48 can form a long-lasting hydrogen bond with E166 of sub unit NthA, which contributes to the expansion of the entrance of substrate access tunnel Caldalkalibacillus thermarum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.84 0.6
-
 3-Cyanopyridine mutant L48H, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 1.2
-
 3-Cyanopyridine wild-type, pH 7.4, 30°C Caldalkalibacillus thermarum

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.84 Caldalkalibacillus thermarum F5L3T2 and F5L3T3 F5L3T2 i.e. subunit NthA, F5L3T3 i.e. subunit NthB
-
4.2.1.84 Caldalkalibacillus thermarum TA2. A1 F5L3T2 and F5L3T3 F5L3T2 i.e. subunit NthA, F5L3T3 i.e. subunit NthB
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.84 217.2
-
 wild-type, substrate 2-cyanopyridine, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 225.9
-
 wild-type, substrate cyanopyrazine, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 383.8
-
 wild-type, substrate 3-cyanopyridine, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 608.2
-
 mutant L48H, substrate 2-cyanopyridine, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 690.5
-
 mutant L48H, substrate cyanopyrazine, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 902.3
-
 mutant L48H, substrate 3-cyanopyridine, pH 7.4, 30°C Caldalkalibacillus thermarum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.84 2-cyanopyridine + H2O
-
 Caldalkalibacillus thermarum pyridine-2-carbamide
-
 ?
4.2.1.84 2-cyanopyridine + H2O
-
 Caldalkalibacillus thermarum TA2. A1 pyridine-2-carbamide
-
 ?
4.2.1.84 3-cyanopyridine + H2O
-
 Caldalkalibacillus thermarum nicotinamide
-
 ?
4.2.1.84 3-cyanopyridine + H2O
-
 Caldalkalibacillus thermarum TA2. A1 nicotinamide
-
 ?
4.2.1.84 cyanopyrazine + H2O
-
 Caldalkalibacillus thermarum pyrazincarbamide
-
 ?
4.2.1.84 cyanopyrazine + H2O
-
 Caldalkalibacillus thermarum TA2. A1 pyrazincarbamide
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.84 352.1
-
 3-Cyanopyridine wild-type, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 833.2
-
 3-Cyanopyridine mutant L48H, pH 7.4, 30°C Caldalkalibacillus thermarum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.1.84 293.4
-
 3-Cyanopyridine wild-type, pH 7.4, 30°C Caldalkalibacillus thermarum
4.2.1.84 1388.7
-
 3-Cyanopyridine mutant L48H, pH 7.4, 30°C Caldalkalibacillus thermarum