Cloned (Comment) | Organism |
---|---|
- |
Mesorhizobium loti |
expressed in Escherichia coli BL21(DE3) cells | Mesorhizobium loti |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, using 8% (w/v) PEG 8000, 0.1 M Tris-HCl, pH 8.5 | Mesorhizobium loti |
structure of enzyme with and without substrates 2-methyl-3-hydroxypyridine-5-carboxylic acid, 5-hydroxynicotinic acid and 5-pyridoxic acid, and of mutant Y270F. Residue Y270 is located in the active site | Mesorhizobium loti |
Protein Variants | Comment | Organism |
---|---|---|
Y270A | the mutant shows reduced ring opening activity but increased NADH oxidation activity compared to the wild type enzyme | Mesorhizobium loti |
Y270A | less than 0.1% of the pyridine ring cleavage capacity of wild-type | Mesorhizobium loti |
Y270F | the mutant shows reduced ring opening activity but increased NADH oxidation activity compared to the wild type enzyme | Mesorhizobium loti |
Y270F | about 1.5% of the pyridine ring cleavage capacity of wild-type | Mesorhizobium loti |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5-hydroxynicotinic acid | - |
Mesorhizobium loti |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
5-hydroxynicotinic acid | mutant enzyme Y270A, at pH 8.0 and 25°C | Mesorhizobium loti | |
0.06 | - |
5-hydroxynicotinic acid | wild type enzyme, at pH 8.0 and 25°C | Mesorhizobium loti | |
0.1 | - |
NADH | wild type enzyme, at pH 8.0 and 25°C | Mesorhizobium loti | |
0.12 | - |
NADH | mutant enzyme Y270F, at pH 8.0 and 25°C | Mesorhizobium loti | |
0.21 | - |
5-hydroxynicotinic acid | mutant enzyme Y270F, at pH 8.0 and 25°C | Mesorhizobium loti | |
0.47 | - |
NADH | mutant enzyme Y270A, at pH 8.0 and 25°C | Mesorhizobium loti |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-methyl-3-hydroxypyridine-5-carboxylic acid + NADH + O2 + H+ | Mesorhizobium loti | - |
2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | Mesorhizobium loti | - |
2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | Mesorhizobium loti MAFF303099 | - |
2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | Mesorhizobium loti MAFF 303099 | - |
2-(acetamidomethylene)succinate + NAD+ | - |
? | |
5-hydroxynicotinic acid + NADH + O2 + H+ | Mesorhizobium loti | about 100% of the activity with 2-methyl-3-hydroxypyridine-5-carboxylic acid | 2-(formylamidomethylene)succinate + NAD+ | - |
? | |
5-pyridoxic acid + NADH + O2 + H+ | Mesorhizobium loti | 5% of the activity with 2-methyl-3-hydroxypyridine-5-carboxylic acid | 2-(acetamidomethylene)-beta-hydroxymethyl succinate + NAD+ | - |
? | |
additional information | Mesorhizobium loti | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | ? | - |
? | |
additional information | Mesorhizobium loti MAFF303099 | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | ? | - |
? | |
additional information | Mesorhizobium loti MAFF 303099 | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mesorhizobium loti | Q988D3 | - |
- |
Mesorhizobium loti MAFF 303099 | Q988D3 | - |
- |
Mesorhizobium loti MAFF303099 | Q988D3 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Mesorhizobium loti |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-methyl-3-hydroxypyridine-5-carboxylic acid + NADH + O2 + H+ | - |
Mesorhizobium loti | 2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti | 2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF303099 | 2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF 303099 | 2-(acetamidomethylene)succinate + NAD+ | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti | 2-(acetamidomethylene)succinate + NAD+ + H2O | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF303099 | 2-(acetamidomethylene)succinate + NAD+ + H2O | - |
? | |
3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF 303099 | 2-(acetamidomethylene)succinate + NAD+ + H2O | - |
? | |
5-hydroxynicotinic acid + NADH + H+ + O2 | - |
Mesorhizobium loti | alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O | - |
? | |
5-hydroxynicotinic acid + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF303099 | alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O | - |
? | |
5-hydroxynicotinic acid + NADH + H+ + O2 | - |
Mesorhizobium loti MAFF 303099 | alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O | - |
? | |
5-hydroxynicotinic acid + NADH + O2 + H+ | about 100% of the activity with 2-methyl-3-hydroxypyridine-5-carboxylic acid | Mesorhizobium loti | 2-(formylamidomethylene)succinate + NAD+ | - |
? | |
5-pyridoxic acid + NADH + H+ + O2 | 5% activity compared to 3-hydroxy-2-methylpyridine-5-carboxylate | Mesorhizobium loti | alpha-(N-acetylaminomethylene)-beta-hydroxymethyl succinic acid + NAD+ + H2O | - |
? | |
5-pyridoxic acid + NADH + H+ + O2 | 5% activity compared to 3-hydroxy-2-methylpyridine-5-carboxylate | Mesorhizobium loti MAFF303099 | alpha-(N-acetylaminomethylene)-beta-hydroxymethyl succinic acid + NAD+ + H2O | - |
? | |
5-pyridoxic acid + NADH + H+ + O2 | 5% activity compared to 3-hydroxy-2-methylpyridine-5-carboxylate | Mesorhizobium loti MAFF 303099 | alpha-(N-acetylaminomethylene)-beta-hydroxymethyl succinic acid + NAD+ + H2O | - |
? | |
5-pyridoxic acid + NADH + O2 + H+ | 5% of the activity with 2-methyl-3-hydroxypyridine-5-carboxylic acid | Mesorhizobium loti | 2-(acetamidomethylene)-beta-hydroxymethyl succinate + NAD+ | - |
? | |
additional information | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | Mesorhizobium loti | ? | - |
? | |
additional information | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | Mesorhizobium loti MAFF303099 | ? | - |
? | |
additional information | also catalyzes the NADH oxidation reaction uncoupled with ring opening in the absence of 2-methyl-3-hydroxypyridine-5-carboxylic acid | Mesorhizobium loti MAFF 303099 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
2-methyl-3-hydroxypyridine 5-carboxylic acid oxygenase | - |
Mesorhizobium loti |
2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase | - |
Mesorhizobium loti |
MHPCO | - |
Mesorhizobium loti |
mlr6788 | - |
Mesorhizobium loti |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
5-hydroxynicotinic acid | mutant enzyme Y270A, at pH 8.0 and 25°C | Mesorhizobium loti | |
19 | - |
5-hydroxynicotinic acid | mutant enzyme Y270F, at pH 8.0 and 25°C | Mesorhizobium loti | |
1300 | - |
5-hydroxynicotinic acid | wild type enzyme, at pH 8.0 and 25°C | Mesorhizobium loti |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Mesorhizobium loti | |
NADH | - |
Mesorhizobium loti |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.48 | - |
wild type enzyme, at pH 8.0 and 25°C | Mesorhizobium loti | 5-hydroxynicotinic acid |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
5-hydroxynicotinic acid | mutant enzyme Y270A, at pH 8.0 and 25°C | Mesorhizobium loti | |
91 | - |
5-hydroxynicotinic acid | mutant enzyme Y270F, at pH 8.0 and 25°C | Mesorhizobium loti | |
22000 | - |
5-hydroxynicotinic acid | wild type enzyme, at pH 8.0 and 25°C | Mesorhizobium loti |