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Literature summary 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)

EC Number Cloned (Comment) Organism
1.14.13.242
-
 Mesorhizobium loti
1.14.13.242 expressed in Escherichia coli BL21(DE3) cells Mesorhizobium loti

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.13.242 hanging drop vapor diffusion method, using 8% (w/v) PEG 8000, 0.1 M Tris-HCl, pH 8.5 Mesorhizobium loti
1.14.13.242 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

EC Number Protein Variants Comment Organism
1.14.13.242 Y270A the mutant shows reduced ring opening activity but increased NADH oxidation activity compared to the wild type enzyme Mesorhizobium loti
1.14.13.242 Y270A less than 0.1% of the pyridine ring cleavage capacity of wild-type Mesorhizobium loti
1.14.13.242 Y270F the mutant shows reduced ring opening activity but increased NADH oxidation activity compared to the wild type enzyme Mesorhizobium loti
1.14.13.242 Y270F about 1.5% of the pyridine ring cleavage capacity of wild-type Mesorhizobium loti

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.242 5-hydroxynicotinic acid
-
 Mesorhizobium loti

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.242 0.045
-
 5-hydroxynicotinic acid mutant enzyme Y270A, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 0.06
-
 5-hydroxynicotinic acid wild type enzyme, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 0.1
-
 NADH wild type enzyme, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 0.12
-
 NADH mutant enzyme Y270F, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 0.21
-
 5-hydroxynicotinic acid mutant enzyme Y270F, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 0.47
-
 NADH mutant enzyme Y270A, at pH 8.0 and 25°C Mesorhizobium loti

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.242 2-methyl-3-hydroxypyridine-5-carboxylic acid + NADH + O2 + H+ Mesorhizobium loti
-
 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 Mesorhizobium loti
-
 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 Mesorhizobium loti MAFF303099
-
 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2 Mesorhizobium loti MAFF 303099
-
 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 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+
-
 ?
1.14.13.242 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+
-
 ?
1.14.13.242 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 ?
-
 ?
1.14.13.242 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 ?
-
 ?
1.14.13.242 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

EC Number Organism UniProt Comment Textmining
1.14.13.242 Mesorhizobium loti Q988D3
-
-
1.14.13.242 Mesorhizobium loti MAFF 303099 Q988D3
-
-
1.14.13.242 Mesorhizobium loti MAFF303099 Q988D3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.242
-
 Mesorhizobium loti

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.242 2-methyl-3-hydroxypyridine-5-carboxylic acid + NADH + O2 + H+
-
 Mesorhizobium loti 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti MAFF303099 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti MAFF 303099 2-(acetamidomethylene)succinate + NAD+
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti 2-(acetamidomethylene)succinate + NAD+ + H2O
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti MAFF303099 2-(acetamidomethylene)succinate + NAD+ + H2O
-
 ?
1.14.13.242 3-hydroxy-2-methylpyridine-5-carboxylate + NADH + H+ + O2
-
 Mesorhizobium loti MAFF 303099 2-(acetamidomethylene)succinate + NAD+ + H2O
-
 ?
1.14.13.242 5-hydroxynicotinic acid + NADH + H+ + O2
-
 Mesorhizobium loti alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O
-
 ?
1.14.13.242 5-hydroxynicotinic acid + NADH + H+ + O2
-
 Mesorhizobium loti MAFF303099 alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O
-
 ?
1.14.13.242 5-hydroxynicotinic acid + NADH + H+ + O2
-
 Mesorhizobium loti MAFF 303099 alpha-(N-formylaminomethylene)succinic acid + NAD+ + H2O
-
 ?
1.14.13.242 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+
-
 ?
1.14.13.242 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
-
 ?
1.14.13.242 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
-
 ?
1.14.13.242 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
-
 ?
1.14.13.242 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+
-
 ?
1.14.13.242 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 ?
-
 ?
1.14.13.242 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 ?
-
 ?
1.14.13.242 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

EC Number Synonyms Comment Organism
1.14.13.242 2-methyl-3-hydroxypyridine 5-carboxylic acid oxygenase
-
 Mesorhizobium loti
1.14.13.242 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase
-
 Mesorhizobium loti
1.14.13.242 MHPCO
-
 Mesorhizobium loti
1.14.13.242 mlr6788
-
 Mesorhizobium loti

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.242 0.05
-
 5-hydroxynicotinic acid mutant enzyme Y270A, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 19
-
 5-hydroxynicotinic acid mutant enzyme Y270F, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 1300
-
 5-hydroxynicotinic acid wild type enzyme, at pH 8.0 and 25°C Mesorhizobium loti

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.242 FAD
-
 Mesorhizobium loti
1.14.13.242 NADH
-
 Mesorhizobium loti

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.14.13.242 0.48
-
 wild type enzyme, at pH 8.0 and 25°C Mesorhizobium loti 5-hydroxynicotinic acid

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.242 1.1
-
 5-hydroxynicotinic acid mutant enzyme Y270A, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 91
-
 5-hydroxynicotinic acid mutant enzyme Y270F, at pH 8.0 and 25°C Mesorhizobium loti
1.14.13.242 22000
-
 5-hydroxynicotinic acid wild type enzyme, at pH 8.0 and 25°C Mesorhizobium loti