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Literature summary for 5.5.1.2 extracted from

  • Halak, S.; Lehtioe, L.; Basta, T.; Buerger, S.; Contzen, M.; Stolz, A.; Goldman, A.
    Structure and function of the 3-carboxy-cis,cis-muconate lactonizing enzyme from the protocatechuate degradative pathway of Agrobacterium radiobacter S2 (2006), FEBS J., 273, 5169-5182.
    View publication on PubMed
Search for more literature for 5.5.1.2

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Agrobacterium tumefaciens

Crystallization (Commentary)

Crystallization (Comment) Organism
recombinant enzyme at 2.2 A resolution. C-terminal domain present in all other members of fumarase II family is missing and enzyme contains an Arg residue as opposed to Trp in the active site. Activation of the carboxylic nucleophile by a hydrophobic environment is not required for lactonization Agrobacterium tumefaciens

Inhibitors

Inhibitors Comment Organism Structure
citrate noncompetitive, localization of binding pocket on the enzyme outside the active site Agrobacterium tumefaciens
isocitrate noncompetitive, localization of binding pocket on the enzyme outside the active site Agrobacterium tumefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.32
-
 2-carboxy-2,5-dihydro-5-oxofuran-2-acetate
-
 Agrobacterium tumefaciens
11.3
-
 3-sulfomuconate
-
 Agrobacterium tumefaciens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37000
-
 x * 37000, SDS-PAGE Agrobacterium tumefaciens

Organism

Organism UniProt Comment Textmining
Agrobacterium tumefaciens
-
-
-
Agrobacterium tumefaciens Q2HNZ1
-
-
Agrobacterium tumefaciens S2 Q2HNZ1
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Agrobacterium tumefaciens

Reaction

Reaction Comment Organism Reaction ID
cis,cis-butadiene-1,2,4-tricarboxylate = 2-carboxy-2,5-dihydro-5-oxofuran-2-acetate activation of the carboxylic nucleophile by a hydrophobic environment is not required for lactonization Agrobacterium tumefaciens
a

Storage Stability

Storage Stability Organism
-20°C, 50 mM Tris/HCl, 100 mM NaCl, 0.5 mM dithiothreitol, completely stable Agrobacterium tumefaciens
20°C, 50 mM Tris/HCl, 100 mM NaCl, 0.5 mM dithiothreitol, 50% loss of activity Agrobacterium tumefaciens
4°C, 50 mM Tris/HCl, 100 mM NaCl, 0.5 mM dithiothreitol, 36 days, completely stable Agrobacterium tumefaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-carboxy-2,5-dihydro-5-oxofuran-2-acetate
-
 Agrobacterium tumefaciens cis,cis-butadiene-1,2,4-tricarboxylate
-
 ?
2-carboxy-2,5-dihydro-5-oxofuran-2-acetate
-
 Agrobacterium tumefaciens S2 cis,cis-butadiene-1,2,4-tricarboxylate
-
 ?
3-sulfomuconate
-
 Agrobacterium tumefaciens 4-sulfomuconolactone + H2O
-
 ?
3-sulfomuconate
-
 Agrobacterium tumefaciens S2 4-sulfomuconolactone + H2O
-
 ?
additional information no substrate: 3-sulfomuconate Agrobacterium tumefaciens ?
-
 ?

Subunits

Subunits Comment Organism
? x * 37000, SDS-PAGE Agrobacterium tumefaciens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1415
-
 2-carboxy-2,5-dihydro-5-oxofuran-2-acetate
-
 Agrobacterium tumefaciens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 7
-
 Agrobacterium tumefaciens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
7.4
-
 isocitrate
-
 Agrobacterium tumefaciens
18
-
 citrate
-
 Agrobacterium tumefaciens