Literature summary for 1.7.1.13 extracted from

Wilding, B.; Winkler, M.; Petschacher, B.; Kratzer, R.; Egger, S.; Steinkellner, G.; Lyskowski, A.; Nidetzky, B.; Gruber, K.; Klempier, N.
Targeting the substrate binding site of E. coli nitrile reductase QueF by modeling, substrate and enzyme engineering (2013), Chemistry, 19, 7007-7012.

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Cloned(Commentary)

Cloned (Comment)	Organism
-	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
construction of homology model and docking studies of natural and non-natural substrates. Residues C190 and D197 play an essential role in the catalytic mechanism	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
C190A	complete loss of activity	Escherichia coli
D197N	complete loss of activity	Escherichia coli
E230Q	about 4% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	Escherichia coli
F228W	about 8% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	Escherichia coli
H229A	about 15% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	Escherichia coli
Q89A	poor activity	Escherichia coli
Q89L	poor activity	Escherichia coli
S90A	about 10% of wild-type activity for natural substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.006	-	NADPH	wild-type, pH 7.5, 25°C	Escherichia coli
0.0061	-	2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	wild-type, pH 7.5, 25°C	Escherichia coli
0.176	-	5-cyanopyrrolo[2,3-d]pyrimidin-4-one	wild-type, pH 7.5, 25°C	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+	Escherichia coli	natural substrate	2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	Q46920	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
3	-	substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant E230Q, pH 7.5, 25°C	Escherichia coli
3.8	-	substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant S90A, pH 7.5, 25°C	Escherichia coli
4.4	-	substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant E230Q, pH 7.5, 25°C	Escherichia coli
8.7	-	substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant F228W, pH 7.5, 25°C	Escherichia coli
10.8	-	substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant F228W, pH 7.5, 25°C	Escherichia coli
12.9	-	substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant S90A, pH 7.5, 25°C	Escherichia coli
14.3	-	substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant H229A, pH 7.5, 25°C	Escherichia coli
17.9	-	substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, mutant H229A, pH 7.5, 25°C	Escherichia coli
90	-	substrate 5-cyanopyrrolo[2,3-d]pyrimidin-4-one, wild-type, pH 7.5, 25°C	Escherichia coli
117	-	substrate 2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one, wild-type, pH 7.5, 25°C	Escherichia coli

Storage Stability

Storage Stability	Organism
-70°C, 1 year, 1% residual activity	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+	natural substrate	Escherichia coli	2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+	-	?
2-amino-5-cyanopyrrolo[2,3-d]pyrimidine + 2 NADPH + 2 H+	analogue of natural substrate, poor substrate	Escherichia coli	2-amino-5-amino-methyl-pyrrolo[2,3-d]pyrimidine + 2 NADP+	-	?
5-cyanopyrrolo[2,3-d]pyrimidin-4-one + 2 NADPH + 2 H+	analogue of natural substrate	Escherichia coli	5-amino-methyl-pyrrolo[2,3-d]pyrimidin-4-one + 2 NADP+	-	?

Synonyms

Synonyms	Comment	Organism
QueF	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.05	-	5-cyanopyrrolo[2,3-d]pyrimidin-4-one	wild-type, pH 7.5, 25°C	Escherichia coli
0.11	-	2-amino-5-cyanopyrrolo[2,3-d]pyrimidin-4-one	wild-type, pH 7.5, 25°C	Escherichia coli
0.142	-	NADPH	wild-type, pH 7.5, 25°C	Escherichia coli

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Release 2023.1 (January 2023)