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Literature summary for 5.4.99.14 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
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Cloned(Commentary)

Cloned (Comment) Organism
-
 Cupriavidus necator

General Stability

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

Inhibitors

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

KM Value [mM]

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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

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

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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]

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

Storage Stability

Storage Stability Organism
4°C or -20°C, highly purified enzyme is rather unstable Cupriavidus necator
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
4-Carboxymethyl-4-methylbut-2-en-1,4-olide
-
 Cupriavidus necator 4-Carboxymethyl-3-methylbut-2-en-1,4-olide
-
 ?
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 ?
-
 ?
additional information not: isomeric methyl-substituted 4-carboxymethylbut-2-en-4-olides Cupriavidus necator ?
-
 ?

Subunits

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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