Any feedback?
Literature summary for 1.7.1.B3 extracted from
- Purification and characetrizatio of NfrA1, a Bacillus subtilis nitro/flavin reductase capable of interacting with the bacterial luciferase (1998), Biosci. Biotechnol. Biochem., 62, 1978-1987 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene ipa-43d, DNA and amino acid sequence determination and analysis, genetic structure and sequence comparison, recombinant overexpression in Escherichia coli J83 | Bacillus subtilis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | enzyme kinetic analysis, overview | Bacillus subtilis | |
| 0.00085 | - |
NADPH | pH 7.0, 23°C, recombinant enzyme, with nitrofurazone | Bacillus subtilis |  |
| 0.0163 | - |
nitrofurazone | pH 7.0, 23°C, recombinant enzyme | Bacillus subtilis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 53000 | - |
recombinant untagged enzyme, gel filtration | Bacillus subtilis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| nitrofurazone + NADPH + H+ | Bacillus subtilis | - |
5-(hydroxyamino)furan-2-carbaldehyde semicarbazone + NADP+ + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus subtilis | - |
- |
- |
| Bacillus subtilis ISW 1214 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme 23fold from Escherichia coli to homogeneity by dialysis, anion exchange chromatography, dialysis, two steps of Blue Sepharose affinity chromatography, and again dialysis | Bacillus subtilis |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| an aromatic amine + 3 NADP+ + 2 H2O = an aromatic nitrate + 3 NADPH + 3 H+ | the enzyme catalysis obeys the ping-pong Bi-Bi kinetic mechanism with substrate FMN as well as substrate nitrofurazone, but upon coupling with the bioluminescent reaction of luciferase, it changes to the sequential mechanism | Bacillus subtilis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 52.5 | - |
purified recombinant untagged enzyme, pH 7.0, 23°C | Bacillus subtilis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| methyl 4-nitrobenzoate + NADPH + H+ | - |
Bacillus subtilis | ? + NADP+ + H2O | - |
? | |
| additional information | the enzyme reduces both nitrofurazone and FMN effectively, see for EC 1.5.1.38, it is bifunctional as flavin reductase and nitroreductase. Two different FMN molecules are involved in the FMN reduction process: FMN tightly associated to the enzyme as prosthetic group and FMN as a substrate. The enzyme is also active with FAD, riboflavin, and lumiflavin, the two latter give the highest activity | Bacillus subtilis | ? | - |
? | |
| nitrofurazone + NADPH + H+ | - |
Bacillus subtilis | 5-(hydroxyamino)furan-2-carbaldehyde semicarbazone + NADP+ + H2O | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homodimer | 2 * 28320, sequence calculation, 2 * 28000, recombinant untagged enzyme, SDS-PAGE | Bacillus subtilis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ipa-43d | - |
Bacillus subtilis |
| More | cf. EC 1.5.1.30 | Bacillus subtilis |
| NADPH-FMN reductase | - |
Bacillus subtilis |
| NADPH-nitrofurazone reductase | - |
Bacillus subtilis |
| NfrA1 | - |
Bacillus subtilis |
| nitro/flavin reductase | - |
Bacillus subtilis |
| non-luminescent-bacterial NfsA/Frp-type enzyme | - |
Bacillus subtilis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 23 | - |
assay at | Bacillus subtilis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Bacillus subtilis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FMN | tightly associated to, two different FMN molecules are involved in the FMN reduction process: FMN tightly associated to the enzyme as prosthetic group and FMN as a substrate | Bacillus subtilis | |
| additional information | NADPH is much more effective than NADH | Bacillus subtilis | |
| NADPH | dependent on | Bacillus subtilis | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | analysis of evolutionary or molecular mechanism of divergence of the nitroreductase/flavin reductase family, overview. The enzyme is similar to NfsA from Escherichia coli, overview | Bacillus subtilis |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
