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Literature summary extracted from

  • Pieper, D.H.; Stadler-Fritzsche, K.; Knackmuss, H.J.; Engesser, K.H.; Bruce, N.C.; Cain, R.B.
    Purification and characterization of 4-methylmuconolactone methylisomerase, a novel enzyme of the modified 3-oxoadipate pathway in the gram-negative bacterium Alcaligenes eutrophus JMP 134 (1990), Biochem. J., 271, 529-534.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.4.99.14
-
 Cupriavidus necator

General Stability

EC Number General Stability Organism
5.4.99.14 Chelating agents like EDTA or thiol-group modifying reagents like DTT stabilize Cupriavidus necator

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.4.99.14 2,4-dimethyl-2-enelactone weak Cupriavidus necator
5.4.99.14 2-Methyl-2-enelactone weak Cupriavidus necator
5.4.99.14 3-methyl-2-enelactone weak Cupriavidus necator
5.4.99.14 4-carboxymethylbut-2-en-4-olide
-
 Cupriavidus necator
5.4.99.14 4-methyl-2-enelactone weak Cupriavidus necator
5.4.99.14 CuSO4
-
 Cupriavidus necator
5.4.99.14 additional information not: isomeric methyl-substituted 4-carboxymethylbut-2-en-4-olides Cupriavidus necator
5.4.99.14 NEM
-
 Cupriavidus necator
5.4.99.14 PCMB
-
 Cupriavidus necator
5.4.99.14 trans-dienelactone weak Cupriavidus necator

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.4.99.14 0.028
-
 1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane
-
 Cupriavidus necator
5.4.99.14 0.176
-
 4-carboxymethyl-4-methylbut-2-en-1,4-olide
-
 Cupriavidus necator

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.4.99.14 40000
-
 1 * 40000, Alcaligenes eutrophus (JMP 134, cloned into Pseudomonas sp. B13 FR1), SDS-PAGE Cupriavidus necator

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.4.99.14 4-Carboxymethyl-4-methylbut-2-en-1,4-olide Cupriavidus necator enzyme of the modified 3-oxoadipate pathway in the Gram-negative bacterium Alcaligenes eutrophus JMP 134 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.14 Cupriavidus necator
-
 basonym Alcaligenes eutrophus
-
5.4.99.14 Cupriavidus necator
-
 JMP 134, cloned into Pseudomonas sp. B13 FR1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.4.99.14 JMP 134, cloned into Pseudomonas sp. B13 FR1 Cupriavidus necator

Reaction

EC Number Reaction Comment Organism Reaction ID
5.4.99.14 4-Carboxymethyl-4-methylbut-2-en-1,4-olide = 4-carboxymethyl-3-methylbut-2-en-1,4-olide mechanism Cupriavidus necator
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.4.99.14 45.2
-
-
 Cupriavidus necator

Storage Stability

EC Number Storage Stability Organism
5.4.99.14 4°C or -20°C, highly purified enzyme is rather unstable Cupriavidus necator
5.4.99.14 4°C, 24 h, 10% loss of activity, highly purified enzyme, 5% loss of activity with addition of chelating agents like EDTA or thiol-group modifying reagents like DTT Cupriavidus necator

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.14 1-Methylbislactone i.e. 1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane Cupriavidus necator 4-Carboxymethyl-3-methylbut-2-en-1,4-olide
-
 ?
5.4.99.14 4-Carboxymethyl-4-methylbut-2-en-1,4-olide
-
 Cupriavidus necator 4-Carboxymethyl-3-methylbut-2-en-1,4-olide
-
 ?
5.4.99.14 4-Carboxymethyl-4-methylbut-2-en-1,4-olide enzyme of the modified 3-oxoadipate pathway in the Gram-negative bacterium Alcaligenes eutrophus JMP 134 Cupriavidus necator ?
-
 ?
5.4.99.14 additional information not: isomeric methyl-substituted 4-carboxymethylbut-2-en-4-olides Cupriavidus necator ?
-
 ?

Subunits

EC Number Subunits Comment Organism
5.4.99.14 monomer 1 * 40000, Alcaligenes eutrophus (JMP 134, cloned into Pseudomonas sp. B13 FR1), SDS-PAGE Cupriavidus necator

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.4.99.14 25
-
 assay at Cupriavidus necator

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.4.99.14 3.75
-
 1-methyl-3,7-dioxo-2,6-dioxabicyclo[3.3.0]octane
-
 Cupriavidus necator
5.4.99.14 98.5
-
 4-carboxymethyl-4-methylbut-2-en-1,4-olide
-
 Cupriavidus necator