Activating Compound | Comment | Organism | Structure |
---|---|---|---|
MetV | the enzyme complex subunit MetV, an iron-sulfur zinc protein, is required by MetF for full activity, and activates the enzyme 70fold with benzyl viologen | Moorella thermoacetica |
Cloned (Comment) | Organism |
---|---|
gene metF, which is part of a transcription unit also containing the genes hdrCBA, mvhD, and metV, genetic structure, recombinant expression of Strep-tagged enzyme MetF in Escherichia coli strain C41(DE3) harboring pCodonPlus and pRKISC, the recombinant MetF contains FMN rather than FAD | Moorella thermoacetica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | the reduction of benzyl viologen with NADH is competitively inhibited by NADP+ | Moorella thermoacetica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
benzyl viologen | pH 7.5, 45°C, recombinant enzyme, with 5-methyltetrahydrofolate | Moorella thermoacetica | |
1.8 | - |
5-methyltetrahydrofolate | pH 7.5, 45°C, recombinant enzyme, with benzyl viologen | Moorella thermoacetica |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Moorella thermoacetica | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | the enzyme contains 110 nmol FAD per mg of enzyme protein | Moorella thermoacetica |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
about, hexaheteromeric complex including MetF | Moorella thermoacetica |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Moorella thermoacetica | - |
- |
- |
Moorella thermoacetica DSM 521 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
native enzyme aerobically 40fold by ultracentrifugation, anion exchange chromatography, gel filtration, ultrafiltration, hydroxyapatite chromatography, followed by again ultrafiltration and gel filtration, enzyme MetF copurifies with the other five proteins encoded in the unit in a hexaheteromeric complex with an apparent molecular mass in the 320-kDa range. Recombinant Strep-tagged enzyme MetF anaerobically from Escherichia coli strain C41(DE3) by avidine affinity chromatography and ultrafiltration | Moorella thermoacetica |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | optimum growth temperature of Moorella thermoacetica is 55°C | Moorella thermoacetica | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
16 | - |
purified enzyme, substrate 5-methyltetrahydrofolate, pH 7.5, 45°C | Moorella thermoacetica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-methyltetrahydrofolate + 2 benzyl viologen | - |
Moorella thermoacetica | 5,10-methylenetetrahydrofolate + 2 reduced benzyl viologen + 2 H+ | - |
r | |
5-methyltetrahydrofolate + 2 benzyl viologen | - |
Moorella thermoacetica DSM 521 | 5,10-methylenetetrahydrofolate + 2 reduced benzyl viologen + 2 H+ | - |
r | |
additional information | the enzyme catalyzes reduction of methylenetetrahydrofolate with reduced benzyl viologen but not with NAD(P)H in either the absence or presence of oxidized ferredoxin. The enzyme also catalyzes the reversible reduction of benzyl viologen with NADH (diaphorase activity) at 48 U/mg | Moorella thermoacetica | ? | - |
? | |
additional information | the enzyme catalyzes reduction of methylenetetrahydrofolate with reduced benzyl viologen but not with NAD(P)H in either the absence or presence of oxidized ferredoxin. The enzyme also catalyzes the reversible reduction of benzyl viologen with NADH (diaphorase activity) at 48 U/mg | Moorella thermoacetica DSM 521 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterohexamer | - |
Moorella thermoacetica |
Synonyms | Comment | Organism |
---|---|---|
metF | - |
Moorella thermoacetica |
methylene-H4F reductase | - |
Moorella thermoacetica |
methylenetetrahydrofolate reductase | - |
Moorella thermoacetica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
- |
Moorella thermoacetica |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 55 | at 55°C the activity is twice as high as at 45°C | Moorella thermoacetica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Moorella thermoacetica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | the enzyme contains 3.1 nmol FAD per mg of enzyme protein. In the absence of FAD, the enzyme is still active, the purified recombinant MetF contains FMN rather than FAD | Moorella thermoacetica | |
FMN | the enzyme contains 3.4 nmol FAD per mg of enzyme protein, the purified recombinant MetF contains FMN rather than FAD | Moorella thermoacetica | |
additional information | The enzyme complex does not catalyze the reduction of methylene-H4F with NADH or NADPH. The enzyme complex subunit HdrA contains iron-sulfur clusters and 2 FADs and catalyzes the reduction of benzyl viologen with NADH. But the physiological electron donor for methylenetetrahydrofolate reduction in Moorella thermoacetica is NADH, and the exergonic reduction of methylenetetrahydrofolate with NADH is coupled via flavin-based electron bifurcation with the endergonic reduction of an yet unknown electron acceptor | Moorella thermoacetica |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in the Wood-Ljungdahl pathway | Moorella thermoacetica |