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Literature summary for 1.5.1.38 extracted from

  • Gwenin, V.; Poornima, P.; Halliwell, J.; Ball, P.; Robinson, G.; Gwenin, C.
    Identification of novel nitroreductases from Bacillus cereus and their interaction with the CB1954 prodrug (2015), Biochem. Pharmacol., 98, 392-402 .
    View publication on PubMed
Search for more literature for 1.5.1.38

Cloned(Commentary)

Cloned (Comment) Organism
gene BC_1619, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain DH5alpha Bacillus cereus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 NADPH with FMN, pH 7.2, 37°C Bacillus cereus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60000
-
 recombinant His-tagged enzyme, gel filtration Bacillus cereus

Organism

Organism UniProt Comment Textmining
Bacillus cereus Q81BW8
-
-
Bacillus cereus ATCC 14579 Q81BW8
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain DH5alpha by nickel affinity chromatography and desalting gel filtration Bacillus cereus

Renatured (Commentary)

Renatured (Comment) Organism
insolubleoverexpressed enzyme protein is resuspended in a resuspension buffer containing 20 ml, 20 mM Tris-HCL, 0.5 M NaCl, pH 8.0, sonicated for 50 s discontinuously, spun down at 8000 rpm (5400 x g) at 4°C for 10 min, pellets resuspended in isolation buffer containing 15 ml, 20 mM Tris-HCL, 0.5 M NaCl, 2% Triton X-100, and 12% w/v urea, pH 8.0 Bacillus cereus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FMNH2 + NADP+ FMN is enzyme-bound Bacillus cereus FMN + NADPH + H+
-
 r
FMNH2 + NADP+ FMN is enzyme-bound Bacillus cereus ATCC 14579 FMN + NADPH + H+
-
 r
additional information the nitroreductase of Bacillus cereus strain ATCC 14579 encoded by gene BC_1619 also shows NADPH-dependent FMN reductase activity, and interacts with the prodrug 5-(1-aziridinyl)-2,4-dinitrobenzamide, i.e. CB1954, converting it to either the toxic 2'- or 4'-hydroxylamine metabolites with cofactor NAD(P)H, overview Bacillus cereus ?
-
 ?
additional information the nitroreductase of Bacillus cereus strain ATCC 14579 encoded by gene BC_1619 also shows NADPH-dependent FMN reductase activity, and interacts with the prodrug 5-(1-aziridinyl)-2,4-dinitrobenzamide, i.e. CB1954, converting it to either the toxic 2'- or 4'-hydroxylamine metabolites with cofactor NAD(P)H, overview Bacillus cereus ATCC 14579 ?
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 26000-34000, SDS-PAGE Bacillus cereus

Synonyms

Synonyms Comment Organism
BC_1619
-
 Bacillus cereus
More cf. EC 1.7.12 Bacillus cereus
ydgI
-
 Bacillus cereus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5
-
 NADPH with FMN, pH 7.2, 37°C Bacillus cereus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
 assay at Bacillus cereus

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Bacillus cereus
NADPH
-
 Bacillus cereus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7.3
-
 NADPH with FMN, pH 7.2, 37°C Bacillus cereus