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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | enzyme kinetic analysis, overview | Bacillus subtilis | |
0.0039 | - |
NADPH | pH 7.0, 23°C, recombinant enzyme, with FMN | Bacillus subtilis | |
0.0047 | - |
FMN | pH 7.0, 23°C, recombinant enzyme | Bacillus subtilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
53000 | - |
recombinant untagged enzyme, gel filtration | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
FAD + NADPH + H+ | Bacillus subtilis | - |
reduced FAD + NADP+ | - |
? | |
FAD + NADPH + H+ | Bacillus subtilis ISW 1214 | - |
reduced FAD + NADP+ | - |
? | |
FMN + NADPH + H+ | Bacillus subtilis | - |
reduced FMN + NADP+ | - |
? | |
FMN + NADPH + H+ | Bacillus subtilis ISW 1214 | - |
reduced FMN + NADP+ | - |
? | |
lumiflavin + NADPH + H+ | Bacillus subtilis | - |
reduced lumiflavin + NADP+ | - |
? | |
lumiflavin + NADPH + H+ | Bacillus subtilis ISW 1214 | - |
reduced lumiflavin + NADP+ | - |
? | |
riboflavin + NADPH + H+ | Bacillus subtilis | - |
reduced riboflavin + NADP+ | - |
? | |
riboflavin + NADPH + H+ | Bacillus subtilis ISW 1214 | - |
reduced riboflavin + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Bacillus subtilis ISW 1214 | - |
- |
- |
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 | Comment | Organism | Reaction ID |
---|---|---|---|
reduced riboflavin + NADP+ = riboflavin + NADPH + 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 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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
FAD + NADPH + H+ | - |
Bacillus subtilis | reduced FAD + NADP+ | - |
? | |
FAD + NADPH + H+ | - |
Bacillus subtilis ISW 1214 | reduced FAD + NADP+ | - |
? | |
FMN + NADPH + H+ | - |
Bacillus subtilis | reduced FMN + NADP+ | - |
? | |
FMN + NADPH + H+ | - |
Bacillus subtilis ISW 1214 | reduced FMN + NADP+ | - |
? | |
lumiflavin + NADPH + H+ | - |
Bacillus subtilis | reduced lumiflavin + NADP+ | - |
? | |
lumiflavin + NADPH + H+ | - |
Bacillus subtilis ISW 1214 | reduced lumiflavin + NADP+ | - |
? | |
additional information | the enzyme reduces both nitrofurazone and FMN effectively, 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 | ? | - |
? | |
additional information | the enzyme reduces both nitrofurazone and FMN effectively, 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 ISW 1214 | ? | - |
? | |
riboflavin + NADPH + H+ | - |
Bacillus subtilis | reduced riboflavin + NADP+ | - |
? | |
riboflavin + NADPH + H+ | - |
Bacillus subtilis ISW 1214 | reduced riboflavin + NADP+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 28320, sequence calculation, 2 * 28000, recombinant untagged enzyme, SDS-PAGE | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
ipa-43d | - |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
23 | - |
assay at | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus subtilis |
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 | 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 |