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Literature summary for 4.2.1.83 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)

Cloned (Comment) Organism
-
Pseudomonas straminea

Protein Variants

Protein Variants Comment Organism
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
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
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
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
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
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
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
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

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Pseudomonas straminea
-
-
-
Pseudomonas straminea NGJ1
-
-
-

Oxidation Stability

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

Purification (Commentary)

Purification (Comment) Organism
recombinant Pseudomonas straminea

Specific Activity [micromol/min/mg]

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

Storage Stability

Storage Stability Organism
-25°C, 20 mM potassium phosphate buffer containing 1 mM DTT, stable for at least 2 months Pseudomonas straminea
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)

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

Subunits

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

Synonyms

Synonyms Comment Organism
4-oxalomesaconate hydratase
-
Pseudomonas straminea
PocOMH
-
Pseudomonas straminea

Turnover Number [1/s]

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