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Literature summary for 1.7.1.B3 extracted from

  • Bai, J.; Yang, J.; Liu, P.; Yang, Q.
    Transformation pathway of 2,4,6-trinitrotoluene by Escherichia coli nitroreductases and improvement of activity using structure-based mutagenesis (2015), Process Biochem., 50, 705-711 .
    View publication on PubMed
Search for more literature for 1.7.1.B3

Cloned(Commentary)

Cloned (Comment) 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 Comment 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, 37°C, recombinant His6-tagged enzyme Escherichia coli
0.3257
-
 2-amino-4,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli
0.6427
-
 4-amino-2,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2,4,6-trinitrotoluene + NADPH + H+ Escherichia coli
-
 4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P17117
-
-

Purification (Commentary)

Purification (Comment) 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4,6-trinitrotoluene + NADPH + H+
-
 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 Escherichia coli 4-hydroxylamino-2,6-dinitrotoluene + NADP+ + H2O
-
 ?
2-amino-4,6-dinitrotoluene + NADPH + H+
-
 Escherichia coli ? + NADP+ + H2O
-
 ?
4-amino-2,6-dinitrotoluene + NADPH + H+
-
 Escherichia coli ? + NADP+ + H2O
-
 ?
additional information no activity with 2,4-diamino-6-nitrotoluene and 2,6-diamino-4-nitrotoluene Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
NfsA
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.05
-
 4-amino-2,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli
12.36
-
 2-amino-4,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli
56.2
-
 2,4,6-trinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADPH
-
 Escherichia coli

General Information

General Information Comment 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 Comment Organism Structure
1.6
-
 4-amino-2,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli
38
-
 2-amino-4,6-dinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli
1931
-
 2,4,6-trinitrotoluene pH 7.0, 37°C, recombinant His6-tagged enzyme Escherichia coli