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show all sequences of 1.7.1.B3

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)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene nfsA, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
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 enzyme
Escherichia coli
0.029
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
0.3257
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
0.6427
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,4,6-trinitrotoluene + NADPH + H+
Escherichia coli
-
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P17117
-
-
Purification (Commentary)
Commentary
Organism
recombinant N-terminally His6-tagged enzyme 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
2,4,6-trinitrotoluene + NADPH + H+
-
743709
Escherichia coli
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
i.e. TNT, NfsA reduces TNT to the para isomer 4-hydroxylamino-2,6-dinitrotoluene more easily than the ortho isomer 2-hydroxylamino-4,6-dinitrotoluene
743709
Escherichia coli
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2-amino-4,6-dinitrotoluene + NADPH + H+
-
743709
Escherichia coli
? + NADP+ + H2O
-
-
-
?
4-amino-2,6-dinitrotoluene + NADPH + H+
-
743709
Escherichia coli
? + NADP+ + H2O
-
-
-
?
additional information
no activity with 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene
743709
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
1.05
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
12.36
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
56.2
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
gene nfsA, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
-
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 enzyme
Escherichia coli
0.029
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
0.3257
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
0.6427
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2,4,6-trinitrotoluene + NADPH + H+
Escherichia coli
-
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His6-tagged enzyme 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
2,4,6-trinitrotoluene + NADPH + H+
-
743709
Escherichia coli
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2,4,6-trinitrotoluene + NADPH + H+
i.e. TNT, NfsA reduces TNT to the para isomer 4-hydroxylamino-2,6-dinitrotoluene more easily than the ortho isomer 2-hydroxylamino-4,6-dinitrotoluene
743709
Escherichia coli
4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
-
-
?
2-amino-4,6-dinitrotoluene + NADPH + H+
-
743709
Escherichia coli
? + NADP+ + H2O
-
-
-
?
4-amino-2,6-dinitrotoluene + NADPH + H+
-
743709
Escherichia coli
? + NADP+ + 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
1.05
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
12.36
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
56.2
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
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 hydroxylamino and/or amino groups on various nitro-substituted compounds
Escherichia coli
additional information
the enzyme crystal structure of NfsA, PDB ID 1F5V, 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 hydroxylamino and/or amino groups on various nitro-substituted compounds
Escherichia coli
additional information
the enzyme crystal structure of NfsA, PDB ID 1F5V, 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
1.6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
38
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
1931
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
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
1.6
-
4-amino-2,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
38
-
2-amino-4,6-dinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
1931
-
2,4,6-trinitrotoluene
pH 7.0, 37C, recombinant His6-tagged enzyme
Escherichia coli
Other publictions for EC 1.7.1.B3
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
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)
741786
Valiauga
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14-22
2017
-
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-
-
-
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-
-
-
-
-
1
-
-
-
1
-
-
-
-
13
-
1
-
-
12
1
-
-
2
2
-
-
-
2
-
2
-
-
-
-
2
2
13
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-
-
-
-
-
-
-
-
-
-
-
13
-
1
-
-
12
1
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-
-
-
2
2
-
12
12
743709
Bai
Transformation pathway of 2,4 ...
Escherichia coli
Process Biochem.
50
705-711
2015
-
-
1
-
-
-
-
4
-
-
-
1
-
1
-
-
1
-
-
-
-
-
5
-
1
-
-
3
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
4
-
-
-
1
-
-
-
1
-
-
-
-
5
-
1
-
-
3
1
-
-
-
-
2
2
-
3
3
742167
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Escherichia coli
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-
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1
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19
-
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1
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4
-
5
1
1
-
-
20
1
-
-
2
-
-
-
-
-
1
2
-
3
-
-
1
-
19
-
-
-
-
-
1
-
1
-
-
4
-
5
1
1
-
-
20
1
-
-
-
-
1
1
-
20
20
743280
Green
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Escherichia coli
Mol. Cancer
12
58
2013
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
742109
Zenno
Purification and characetriza ...
Bacillus subtilis, Bacillus subtilis ISW 1214
Biosci. Biotechnol. Biochem.
62
1978-1987
1998
-
-
1
-
-
-
-
3
-
-
1
1
-
2
-
-
1
1
-
-
1
-
3
1
1
-
-
-
1
-
-
3
-
-
-
-
-
1
3
-
-
-
-
-
-
3
-
-
1
1
-
-
-
1
-
-
1
-
3
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-