Cloned (Comment) | Organism |
---|---|
overexpression of hemF oxygen-dependent enzyme form as N-terminally fusion protein with calmodulin-binding-peptide, overexpression of wild-type and mutant enzymes in strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C167S | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
G127V | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
H106L | site-directed mutagenesis, inactive mutant | Escherichia coli |
H145L | site-directed mutagenesis, inactive mutant | Escherichia coli |
H175L | site-directed mutagenesis, inactive mutant | Escherichia coli |
H96L | site-directed mutagenesis, inactive mutant | Escherichia coli |
additional information | mutation of the nucleotide binding motif GGGXXTP does not affect the enzyme activity | Escherichia coli |
P133A | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
T132A | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
W123L | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
W124R | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
W166L | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
W274L | site-directed mutagenesis, inactive mutant | Escherichia coli |
W298L | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
W36L | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Y135F | site-directed mutagenesis, activity similar to the wild-type | Escherichia coli |
Y160F | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Y170F | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Y213F | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Y240F | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Y276F | site-directed mutagenesis, activity and kinetics similar to the wild-type | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | - |
Escherichia coli | |
EGTA | - |
Escherichia coli | |
additional information | mutation of the nucleotide binding motif GGGXXTP does not affect the enzyme activity | Escherichia coli | |
protoporphyrin-IX | inhibition of HemF | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0016 | - |
Coproporphyrinogen-III | HemF mutant C167S | Escherichia coli | |
0.002 | - |
Coproporphyrinogen-III | HemF mutant Y240F | Escherichia coli | |
0.0021 | - |
Coproporphyrinogen-III | HemF mutant G127V | Escherichia coli | |
0.0026 | - |
Coproporphyrinogen-III | recombinant O2-dependent enzyme form HemF | Escherichia coli | |
0.003 | - |
Coproporphyrinogen-III | HemF mutants Y213F, T132A, and W166L | Escherichia coli | |
0.0032 | - |
Coproporphyrinogen-III | HemF mutant W298L | Escherichia coli | |
0.0033 | - |
Coproporphyrinogen-III | HemF mutant W123L | Escherichia coli | |
0.0035 | - |
Coproporphyrinogen-III | HemF mutants Y160F, W123L, and P133A | Escherichia coli | |
0.0036 | - |
Coproporphyrinogen-III | HemF mutant W124R | Escherichia coli | |
0.0038 | - |
Coproporphyrinogen-III | HemF mutant Y135F | Escherichia coli | |
0.0041 | - |
Coproporphyrinogen-III | HemF mutant Y170F | Escherichia coli | |
0.0043 | - |
Coproporphyrinogen-III | HemF mutants Y276F and W36L | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required, bound to the enzyme, stimulates, restores the enzyme inactivated by metal chelators, coordinated by residues His96, His106, His145, and His175 | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
coproporphyrinogen-III + O2 | Escherichia coli | step in heme biosynthesis | protoporphyrinogen-IX + CO2 + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
oxygen-dependent and oxygen-independent enzymes, HemF and HemN | - |
Purification (Comment) | Organism |
---|---|
recombinant hemF oxygen-dependent enzyme form to homogeneity by affinity chromatography, the tag protein is cleaved off by thrombin, recombinant wild-type and mutant enzymes | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
coproporphyrinogen III + O2 + 2 H+ = protoporphyrinogen-IX + 2 CO2 + 2 H2O | 2 possible catalytic pathways of the reaction, overview, Tyrp274 is catalytically important, the following residues are not important for enzyme activity: Cys167, Tyr135, Tyr160, Tyr170, Tyr213, Tyr240, Tyr276, Trp36, Trp123, Trp166, and Trp198 | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
coproporphyrinogen-III + O2 | step in heme biosynthesis | Escherichia coli | protoporphyrinogen-IX + CO2 + H2O | - |
? | |
coproporphyrinogen-III + O2 | HemF is absolutely dependent on oxygen | Escherichia coli | protoporphyrinogen-IX + CO2 + H2O | - |
? | |
additional information | no activity of HemF with protoporphyrinogen-IX and coporphyrinogen-III | Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
coproporphyrinogen III oxidase | - |
Escherichia coli |
CPO | - |
Escherichia coli |
HemF | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
Coproporphyrinogen-III | recombinant O2-dependent enzyme form | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
recombinant O2-dependent enzyme form | Escherichia coli |