EC Number | Cloned (Comment) | Organism |
---|---|---|
1.5.1.20 | expression of wild-type and mutant enzymes in Escherichia coli strain AB109 and AB1909-(DE3)7D | Escherichia coli |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.5.1.20 | D120A | site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction | Escherichia coli |
1.5.1.20 | D120K | site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction | Escherichia coli |
1.5.1.20 | D120N | site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction | Escherichia coli |
1.5.1.20 | D120S | site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction | Escherichia coli |
1.5.1.20 | D120V | site-directed mutagenesis, decreased catalytic efficiency in the folate oxidative half-reaction, loss of negative charge near the flavin, small increasing effects on the NADH reductive half reaction | Escherichia coli |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.5.1.20 | methylenetetrahydrofolate | - |
Escherichia coli | |
1.5.1.20 | NADH | - |
Escherichia coli |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.5.1.20 | additional information | - |
additional information | redox potentials of wild-type and mutant enzymes, kinetic mechanism, and rapid-reaction kinetics for the half-reactions, steady-state kinetics | Escherichia coli | |
1.5.1.20 | 0.0004 | - |
methylenetetrahydrofolate | recombinant wild-type enzyme, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.0035 | - |
NADH | recombinant wild-type enzyme, pH 7.2, 4°C, the mutant enzymes all show a Km below 0.0035 mM | Escherichia coli | |
1.5.1.20 | 0.017 | - |
methylenetetrahydrofolate | recombinant mutant D120N, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.043 | - |
methylenetetrahydrofolate | recombinant mutant D120S, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.099 | - |
methylenetetrahydrofolate | recombinant mutant D120A, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.142 | - |
methylenetetrahydrofolate | recombinant mutant D120K, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.187 | - |
methylenetetrahydrofolate | recombinant mutant D120V, pH 7.2, 4°C | Escherichia coli |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.5.1.20 | 5-methyltetrahydrofolate + NAD+ | Escherichia coli | - |
5,10-methylenetetrahydrofolate + NADH | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.5.1.20 | Escherichia coli | - |
- |
- |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.5.1.20 | 5-methyltetrahydrofolate + NAD(P)+ = 5,10-methylenetetrahydrofolate + NAD(P)H + H+ | substrate binding and catalyic mechanism involve Asp120, half-reaction mechanisms | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.5.1.20 | (6R,S)-5,10-methylenetetrahydrofolate + ? | - |
Escherichia coli | ? | - |
? | |
1.5.1.20 | 5-methyltetrahydrofolate + NAD+ | - |
Escherichia coli | 5,10-methylenetetrahydrofolate + NADH | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.5.1.20 | methylenetetrahydrofolate reductase | - |
Escherichia coli |
1.5.1.20 | MTHFR | - |
Escherichia coli |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.5.1.20 | 0.007 | - |
methylenetetrahydrofolate | recombinant mutant D120N, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.011 | - |
methylenetetrahydrofolate | recombinant mutant D120K, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.017 | - |
methylenetetrahydrofolate | recombinant mutant D120V, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.02 | - |
methylenetetrahydrofolate | recombinant mutant D120S, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 0.022 | - |
methylenetetrahydrofolate | recombinant mutant D120A, pH 7.2, 4°C | Escherichia coli | |
1.5.1.20 | 2.2 | - |
methylenetetrahydrofolate | recombinant wild-type enzyme, pH 7.2, 4°C | Escherichia coli |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.5.1.20 | 7.2 | - |
assay at | Escherichia coli |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.5.1.20 | FAD | redox properties of enzyme-bound FAD are influenced by Asp120, which electrostatically stabilizes putative 5-iminium cation intermediate during catalysis | Escherichia coli | |
1.5.1.20 | NAD+ | - |
Escherichia coli | |
1.5.1.20 | NADH | - |
Escherichia coli |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.5.1.20 | 0.014 | - |
NADH | recombinant wild-type enzyme, pH 7.2, 4°C, the mutant enzymes all show a Km below 0.0035 mM | Escherichia coli | |
1.5.1.20 | 0.061 | - |
methylenetetrahydrofolate | recombinant wild-type enzyme, pH 7.2, 4°C | Escherichia coli |