Literature summary for 4.2.1.84 extracted from

Ma, D.; Cheng, Z.; Peplowski, L.; Han, L.; Xia, Y.; Hou, X.; Guo, J.; Yin, D.; Rao, Y.; Zhou, Z.
Insight into the broadened substrate scope of nitrile hydratase by static and dynamic structure analysis (2022), Chem. Sci., 13, 8417-8428.

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Protein Variants

Protein Variants	Comment	Organism
A129R	mutation in subunit beta, mutant exhibits an expanded substrate scope, especially toward bulky nitrile substrates. The amino acid residue located at the entrance of the substrate access tunnel is a key factor in enzyme catalytic performance	Pseudonocardia thermophila
M46R	mutation in subunit beta, mutant exhibits an expanded substrate scope, especially toward bulky nitrile substrates. The amino acid residue located at the entrance of the substrate access tunnel is a key factor in enzyme catalytic performance	Pseudonocardia thermophila
M46R/A129R	mutations in subunit beta, double mutant exhibits lower activity than the two single mutants	Pseudonocardia thermophila

Organism

Organism	UniProt	Comment	Textmining
Pseudonocardia thermophila	Q7SID2 and Q7SID3	Q7SID2 i.e. subunit alpha, Q7SID3 i.e. subunit beta	-
Pseudonocardia thermophila JCM 3095	Q7SID2 and Q7SID3	Q7SID2 i.e. subunit alpha, Q7SID3 i.e. subunit beta	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
5.4	-	wild-type, substrate thiacloprid, pH 7.4, 25°C	Pseudonocardia thermophila
13.1	-	wild-type, substrate 1-naphthonitrile, pH 7.4, 25°C	Pseudonocardia thermophila
14.4	-	wild-type, substrate pentanenitrile, pH 7.4, 25°C	Pseudonocardia thermophila
45.8	-	wild-type, substrate cinnamonitrile, pH 7.4, 25°C	Pseudonocardia thermophila
66.7	-	wild-type, substrate 3-cyanopyrazine, pH 7.4, 25°C	Pseudonocardia thermophila
176	-	wild-type, substrate benzonitrile, pH 7.4, 25°C	Pseudonocardia thermophila
203	-	wild-type, substrate 2-cyanopyrazine, pH 7.4, 25°C	Pseudonocardia thermophila
666	-	wild-type, substrate isobutyronitrile, pH 7.4, 25°C	Pseudonocardia thermophila
1982	-	wild-type, substrate hexanenitrile, pH 7.4, 25°C	Pseudonocardia thermophila
2463	-	wild-type, substrate acrylonitrile, pH 7.4, 25°C	Pseudonocardia thermophila

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
1-naphthonitrile + H2O	-	Pseudonocardia thermophila	naphthalene-1-carboxamide	-	?
2-cyanopyrazine + H2O	-	Pseudonocardia thermophila	pyrazincarbamide	-	?
2-cyanopyrazine + H2O	-	Pseudonocardia thermophila JCM 3095	pyrazincarbamide	-	?
3-cyanopyridine + H2O	-	Pseudonocardia thermophila	nicotinamide	-	?
acrylonitrile + H2O	-	Pseudonocardia thermophila	2-propenoic acid amide	-	?
acrylonitrile + H2O	-	Pseudonocardia thermophila JCM 3095	2-propenoic acid amide	-	?
benzonitrile + H2O	-	Pseudonocardia thermophila	benzamide	-	?
cinnamonitrile + H2O	-	Pseudonocardia thermophila	(2E)-3-phenylprop-2-enamide	-	?
hexanenitrile + H2O	-	Pseudonocardia thermophila	hexanamide	-	?
hexanenitrile + H2O	-	Pseudonocardia thermophila JCM 3095	hexanamide	-	?
isobutyronitrile + H2O	-	Pseudonocardia thermophila	2-methylpropanamide	-	?
isobutyronitrile + H2O	-	Pseudonocardia thermophila JCM 3095	2-methylpropanamide	-	?
pentanenitril + H2O	-	Pseudonocardia thermophila	pentanamide	-	?
pentanenitril + H2O	-	Pseudonocardia thermophila JCM 3095	pentanamide	-	?
thiacloprid + H2O	-	Pseudonocardia thermophila	N-[3-[(6-chloropyridin-3-yl)methyl]-1,3-thiazolidin-2-ylidene]urea	-	?

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Release 2026.1 (March 2026)