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Literature summary for 1.5.7.1 extracted from

  • Mock, J.; Wang, S.; Huang, H.; Kahnt, J.; Thauer, R.K.
    Evidence for a hexaheteromeric methylenetetrahydrofolate reductase in Moorella thermoacetica (2014), J. Bacteriol., 196, 3303-3314 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.7.1

Activating Compound

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(Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Moorella thermoacetica 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ the enzyme contains 110 nmol FAD per mg of enzyme protein Moorella thermoacetica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
 about, hexaheteromeric complex including MetF Moorella thermoacetica

Organism

Organism UniProt Comment Textmining
Moorella thermoacetica
-
-
-
Moorella thermoacetica DSM 521
-
-
-

Purification (Commentary)

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

Source Tissue Comment Organism Textmining
additional information optimum growth temperature of Moorella thermoacetica is 55°C Moorella thermoacetica
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
heterohexamer
-
 Moorella thermoacetica

Synonyms

Synonyms Comment Organism
metF
-
 Moorella thermoacetica
methylene-H4F reductase
-
 Moorella thermoacetica
methylenetetrahydrofolate reductase
-
 Moorella thermoacetica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
 Moorella thermoacetica

Temperature Range [°C]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Moorella thermoacetica

Cofactor

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

General Information Comment Organism
metabolism the enzyme is involved in the Wood-Ljungdahl pathway Moorella thermoacetica