BRENDA - Enzyme Database
show all sequences of 1.7.1.B2

Transformation pathway of 2,4,6-trinitrotoluene by Escherichia coli nitroreductases and improvement of activity using structure-based mutagenesis

Bai, J.; Yang, J.; Liu, P.; Yang, Q.; Process Biochem. 50, 705-711 (2015) View publication on PubMed

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene nfsB, recombinant expression of wild-type and mutant N-terminally His6-tagged enzymes in Escherichia coli strain BL21(DE3)
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
F123A
site-directed mutagenesis, residue F123 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124A
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124N
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124W
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
N71S
site-directed mutagenesis, active site residue mutation, the mutant shows increased activity with prodrug 5-[aziridin-1-yl]-2,4-dinitrobenzamide (CB1954) and nitrofurazone compared to the wild-type enzyme
Escherichia coli
T41L
site-directed mutagenesis, active site residue mutation, the mutant shows increased activity with prodrug 5-[aziridin-1-yl]-2,4-dinitrobenzamide (CB1954) and nitrofurazone compared to the wild-type enzyme
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten steady-state kinetics of recombinant His6-tagged wild-type and mutant enzymes
Escherichia coli
0.0023
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.0057
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.0072
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.0091
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.0174
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.0398
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.0534
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
0.0881
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.0928
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
0.0996
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.0997
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
0.163
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.172
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.265
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.306
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.339
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.38
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.411
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
0.518
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.575
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
0.732
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
1.069
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
1.168
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
1.495
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2,4,6-trinitrotoluene + NADH + H+
Escherichia coli
-
4-hydroxyamino-2,6-dinitrotoluene + NAD+ + H2O
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
Escherichia coli
-
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
P38489
-
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant wild-type and mutant N-terminally His6-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to over 95% purity
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2,4,6-trinitrotoluene + NADH + H+
-
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NAD+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
-
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
i.e. TNT, NfsB reduces TNT to the para isomer 4-hydroxyamino-2,6-dinitrotoluene more easily than the ortho isomer 2-hydroxyamino-4,6-dinitrotoluene
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2-amino-4,6-dinitrotoluene + NADH + H+
-
743709
Escherichia coli
4-hydroxyamino-2-aminotoluene + NAD+ + H2O
-
-
-
?
4-amino-2,6-dinitrotoluene + NADH + H+
-
743709
Escherichia coli
2-hydroxyamino-4-aminotoluene + NAD+ + H2O
-
-
-
?
additional information
no activity with 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene
743709
Escherichia coli
?
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
NfsB
-
Escherichia coli
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.76
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1.03
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1.51
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
3.25
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
4.74
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
8.06
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
9.28
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
9.4
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
11.29
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
11.48
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
14.01
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
14.38
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
15.27
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
15.31
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
18.02
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
18.2
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
26.84
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
26.9
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
37.65
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
38.88
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
46.67
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
54.52
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
62.87
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
69.26
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
additional information
NfsB is capable of utilizing bothNADH and NADPH as cofactor
Escherichia coli
NADH
-
Escherichia coli
NADPH
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
gene nfsB, recombinant expression of wild-type and mutant N-terminally His6-tagged enzymes in Escherichia coli strain BL21(DE3)
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
additional information
NfsB is capable of utilizing bothNADH and NADPH as cofactor
Escherichia coli
NADH
-
Escherichia coli
NADPH
-
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
F123A
site-directed mutagenesis, residue F123 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124A
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124N
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
F124W
site-directed mutagenesis, residue F124 is located in helix alpha6 constituting the substrate entry site, the mutant enzyme kinetics are altered compared to the wild-type enzyme
Escherichia coli
N71S
site-directed mutagenesis, active site residue mutation, the mutant shows increased activity with prodrug 5-[aziridin-1-yl]-2,4-dinitrobenzamide (CB1954) and nitrofurazone compared to the wild-type enzyme
Escherichia coli
T41L
site-directed mutagenesis, active site residue mutation, the mutant shows increased activity with prodrug 5-[aziridin-1-yl]-2,4-dinitrobenzamide (CB1954) and nitrofurazone compared to the wild-type enzyme
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten steady-state kinetics of recombinant His6-tagged wild-type and mutant enzymes
Escherichia coli
0.0023
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.0057
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.0072
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.0091
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.0174
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.0398
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.0534
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
0.0881
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.0928
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
0.0996
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.0997
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
0.163
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.172
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
0.265
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
0.306
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
0.339
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.38
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
0.411
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
0.518
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
0.575
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
0.732
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
1.069
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
1.168
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
1.495
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2,4,6-trinitrotoluene + NADH + H+
Escherichia coli
-
4-hydroxyamino-2,6-dinitrotoluene + NAD+ + H2O
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
Escherichia coli
-
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type and mutant N-terminally His6-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to over 95% purity
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2,4,6-trinitrotoluene + NADH + H+
-
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NAD+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
-
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
i.e. TNT, NfsB reduces TNT to the para isomer 4-hydroxyamino-2,6-dinitrotoluene more easily than the ortho isomer 2-hydroxyamino-4,6-dinitrotoluene
743709
Escherichia coli
4-hydroxyamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2-amino-4,6-dinitrotoluene + NADH + H+
-
743709
Escherichia coli
4-hydroxyamino-2-aminotoluene + NAD+ + H2O
-
-
-
?
4-amino-2,6-dinitrotoluene + NADH + H+
-
743709
Escherichia coli
2-hydroxyamino-4-aminotoluene + NAD+ + H2O
-
-
-
?
additional information
no activity with 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene
743709
Escherichia coli
?
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.76
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1.03
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1.51
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
3.25
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
4.74
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
8.06
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
9.28
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
9.4
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
11.29
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
11.48
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
14.01
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
14.38
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
15.27
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
15.31
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
18.02
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
18.2
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
26.84
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
26.9
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
37.65
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
38.88
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
46.67
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
54.52
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
62.87
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
69.26
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Escherichia coli
General Information
General Information
Commentary
Organism
evolution
nitroreductases are a family of flavoproteins that catalyze the NAD(P)H-dependent reduction of nitro groups to hydroxyamino and/or amino groups on vari-ous nitro-substituted compounds
Escherichia coli
additional information
the enzyme crystal structure of NfsB, PDB ID 1DS7, is used for molecular ligand docking studies
Escherichia coli
General Information (protein specific)
General Information
Commentary
Organism
evolution
nitroreductases are a family of flavoproteins that catalyze the NAD(P)H-dependent reduction of nitro groups to hydroxyamino and/or amino groups on vari-ous nitro-substituted compounds
Escherichia coli
additional information
the enzyme crystal structure of NfsB, PDB ID 1DS7, is used for molecular ligand docking studies
Escherichia coli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
11
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
12
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
26
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
27
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
30
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
31
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
31
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
45
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
46
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
57
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
65
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
181
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
206
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
290
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
547
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
653
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1177
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
2170
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
2526
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
5111
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
5389
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
11
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
12
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
26
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
27
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
30
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
31
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
31
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
45
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
46
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
57
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
65
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
181
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
206
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
290
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124N, with NADH
Escherichia coli
547
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124A, with NADH
Escherichia coli
653
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant N71S, with NADH
Escherichia coli
1177
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant T41L, with NADH
Escherichia coli
2170
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged wild-type enzyme, with NADH
Escherichia coli
2526
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F124W, with NADH
Escherichia coli
5111
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A, with NADH
Escherichia coli
5389
-
2,4,6-trinitrotoluene
pH 7.0, 37°C, recombinant His6-tagged mutant F123A/F124W, with NADH
Escherichia coli
Other publictions for EC 1.7.1.B2
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
743709
Bai
Transformation pathway of 2,4 ...
Escherichia coli
Process Biochem.
50
705-711
2015
-
-
1
-
6
-
-
25
-
-
-
2
-
1
-
-
1
-
-
-
-
-
6
-
1
1
-
-
24
1
-
-
3
-
-
-
-
-
1
3
-
6
-
-
-
-
25
-
-
-
2
-
-
-
1
-
-
-
-
6
-
1
-
-
24
1
-
-
-
-
2
2
-
24
24
743280
Green
Pseudomonas aeruginosa NfsB a ...
Escherichia coli, Pseudomonas aeruginosa
Mol. Cancer
12
58
2013
-
1
2
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
8
-
4
-
-
-
-
-
-
-
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-