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Literature summary extracted from

  • Li, S.; Kimura, M.; Takashima, T.; Hayashi, K.; Inoue, K.; Ishiguro, R.; Sugisaki, H.; Maruyama, K.
    Role of cysteine residues in 4-oxalomesaconate hydratase from Pseudomonas ochraceae NGJ1 (2007), Biosci. Biotechnol. Biochem., 71, 449-457.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.83
-
Pseudomonas straminea

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.83 C124S kcat/Km for 4-oxalomesaconate is 1.9fold higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C131S kcat/Km for 4-oxalomesaconate is 26% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C183S kcat/Km for 4-oxalomesaconate is 1.8fold higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C186S kcat/Km for 4-oxalomesaconate is 16% higher than wild-type value, mutant enzyme is more stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C205S kcat/Km for 4-oxalomesaconate is 29% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C260S kcat/Km for 4-oxalomesaconate is 18% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C92S kcat/Km for 4-oxalomesaconate is 18% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea
4.2.1.83 C96S kcat/Km for 4-oxalomesaconate is 3.9fold lower than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT Pseudomonas straminea

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.83 5,5'-dithiobis(2-nitrobenzoic acid) inhibition is primarily due to modification of Cys186. DTNB-treated C186S remains fully active Pseudomonas straminea
4.2.1.83 HgCl2 1 mM, all the mutant enzymes lose activity with incubation (C92S, C96S, C124S, C131S, C183S, C186S, C205S, C260S) Pseudomonas straminea
4.2.1.83 Zn2+
-
Pseudomonas straminea

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.83 0.0029
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C92S Pseudomonas straminea
4.2.1.83 0.00618
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C205S Pseudomonas straminea
4.2.1.83 0.00733
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C260S Pseudomonas straminea
4.2.1.83 0.00794
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C124S Pseudomonas straminea
4.2.1.83 0.00838
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C131S Pseudomonas straminea
4.2.1.83 0.00851
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C183S Pseudomonas straminea
4.2.1.83 0.00994
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C186S Pseudomonas straminea
4.2.1.83 0.0107
-
4-oxalomesaconate 25°C, pH 8.0, wild-type enzyme Pseudomonas straminea
4.2.1.83 0.087
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C96S Pseudomonas straminea

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.83 Zn2+ PocOMH possesses one Zn2+ per subunit. It strongly binds to the enzyme protein Pseudomonas straminea

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.83 38000
-
2 * 38000, SDS-PAGE Pseudomonas straminea
4.2.1.83 72000
-
gel filtration Pseudomonas straminea

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.83 Pseudomonas straminea
-
-
-
4.2.1.83 Pseudomonas straminea NGJ1
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
4.2.1.83 the enzyme is unstable in absence of reducing reagents Pseudomonas straminea

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.83 recombinant Pseudomonas straminea

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.83 113
-
-
Pseudomonas straminea

Storage Stability

EC Number Storage Stability Organism
4.2.1.83 -25°C, 20 mM potassium phosphate buffer containing 1 mM DTT, stable for at least 2 months Pseudomonas straminea
4.2.1.83 4°C, 20 mM potassium phosphate buffer containing 1 mM DTT, wild-type enzyme loses 20% of initial activity after 1 d Pseudomonas straminea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.83 4-oxalomesaconate
-
Pseudomonas straminea ?
-
?
4.2.1.83 4-oxalomesaconate
-
Pseudomonas straminea NGJ1 ?
-
?

Subunits

EC Number Subunits Comment Organism
4.2.1.83 dimer 2 * 38000, SDS-PAGE Pseudomonas straminea

Synonyms

EC Number Synonyms Comment Organism
4.2.1.83 4-oxalomesaconate hydratase
-
Pseudomonas straminea
4.2.1.83 PocOMH
-
Pseudomonas straminea

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.83 2.2
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C131S Pseudomonas straminea
4.2.1.83 2.3
-
4-oxalomesaconate 25°C, pH 8.0, wild-type enzyme Pseudomonas straminea
4.2.1.83 24.1
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C92S Pseudomonas straminea
4.2.1.83 33.9
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C183S Pseudomonas straminea
4.2.1.83 56
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C205S Pseudomonas straminea
4.2.1.83 60.8
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C260S Pseudomonas straminea
4.2.1.83 74.1
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C131S Pseudomonas straminea
4.2.1.83 75.4
-
4-oxalomesaconate 25°C, pH 8.0, wild-type enzyme Pseudomonas straminea
4.2.1.83 81.3
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C186S Pseudomonas straminea
4.2.1.83 104
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C124S Pseudomonas straminea
4.2.1.83 159
-
4-oxalomesaconate 25°C, pH 8.0, mutant enzyme C96S Pseudomonas straminea