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Literature summary for 1.14.13.242 extracted from

  • Kobayashi, J.; Yoshida, H.; Yagi, T.; Kamitori, S.; Hayashi, H.; Mizutani, K.; Takahashi, N.; Mikami, B.
    Role of the Tyr270 residue in 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase from Mesorhizobium loti (2017), J. Biosci. Bioeng., 123, 154-162 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
-
Mesorhizobium loti
expressed in Escherichia coli BL21(DE3) cells Mesorhizobium loti

Crystallization (Commentary)

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

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

Inhibitors Comment Organism Structure
5-hydroxynicotinic acid
-
Mesorhizobium loti

KM Value [mM]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Mesorhizobium loti Q988D3
-
-
Mesorhizobium loti MAFF 303099 Q988D3
-
-
Mesorhizobium loti MAFF303099 Q988D3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Mesorhizobium loti

Substrates and Products (Substrate)

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

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 [1/s]

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

Cofactor Comment Organism Structure
FAD
-
Mesorhizobium loti
NADH
-
Mesorhizobium loti

IC50 Value

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 [mM/s]

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