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Literature summary extracted from

  • Thompson, L.C.; Ladner, J.E.; Codreanu, S.G.; Harp, J.; Gilliland, G.L.; Armstrong, R.N.
    2-Hydroxychromene-2-carboxylic acid isomerase: a kappa class glutathione transferase from Pseudomonas putida (2007), Biochemistry, 46, 6710-6722.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.18 expression in Escherichia coli strain BL21(DE3) Pseudomonas putida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.18 purified enzyme complexed with glutathione, 2-hydroxychromene-2-carboxylic acid, and trans-o-hydroxybenzylidene pyruvic acid, 10 mg/ml protein in storage buffer, with 1 mM GSH, and 1 mM HCCA at 21°C, all crystals are grown under 7 ml of Al’s oil in a microbatch plate, X-ray diffraction structure determination and analysis at 1.7 A resolution, structure modeling Pseudomonas putida
5.99.1.4 crystal structure of the enzyme at 1.7 A resolution Pseudomonas putida

General Stability

EC Number General Stability Organism
5.99.1.4 unstable in absence of GSH Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.18 trans-o-hydroxybenzylidene pyruvic acid
-
 Pseudomonas putida
5.99.1.4 (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate i.e. (E)-2'-hydroxybenzylidenepyruvate Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.18 additional information
-
 additional information steady-state and ligand binding kinetics, recombinant enzyme Pseudomonas putida
2.5.1.18 0.017
-
 reduced glutathione pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
2.5.1.18 0.084
-
 2-hydroxychromene-2-carboxylic acid pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
2.5.1.18 0.138
-
 trans-o-hydroxybenzylidene pyruvic acid pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
5.99.1.4 0.084
-
 2-hydroxychromene-2-carboxylate pH 7.0, 25°C Pseudomonas putida
5.99.1.4 0.138
-
 (E)-2'-hydroxybenzylidenepyruvate pH 7.0, 25°C Pseudomonas putida

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.5.1.18 soluble
-
 Pseudomonas putida
-
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.18 2-hydroxychromene-2-carboxylic acid Pseudomonas putida HCCA isomerase catalyzes the glutathione-dependent interconversion of 2-hydroxychromene-2-carboxylic acid and trans-o-hydroxybenzylidene pyruvic acid in the naphthalene catabolic pathway of Pseudomonas putida, mechanisms in the naphthalene catabolic pathwayoverview trans-o-hydroxybenzylidene pyruvic acid
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.18 Pseudomonas putida
-
-
-
5.99.1.4 Pseudomonas putida Q51948
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.18 recombinant soluble enzyme from supernatant by anion exchange chromatograpy and gel filtration Pseudomonas putida

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.18 270
-
 purified recombinant enzyme in presence of reduced glutathione Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.18 2-hydroxychromene-2-carboxylic acid HCCA isomerase catalyzes the glutathione-dependent interconversion of 2-hydroxychromene-2-carboxylic acid and trans-o-hydroxybenzylidene pyruvic acid in the naphthalene catabolic pathway of Pseudomonas putida, mechanisms in the naphthalene catabolic pathwayoverview Pseudomonas putida trans-o-hydroxybenzylidene pyruvic acid
-
 ?
2.5.1.18 2-hydroxychromene-2-carboxylic acid the dimeric protein binds one molecule of GSH very tightly and a second molecule of GSH with much lower affinity, GSH acts as a tightly bound cofactor in the reaction, binding structure, overview Pseudomonas putida trans-o-hydroxybenzylidene pyruvic acid
-
 ?
2.5.1.18 2-hydroxychromene-2-carboxylic acid + reduced glutathione
-
 Pseudomonas putida trans-o-hydroxybenzylidene pyruvic acid + ?
-
 ?
5.99.1.4 2-hydroxy-2H-chromene-2-carboxylate glutathione (GSH)-dependent interconversion. The isomerization reaction involves a short-lived covalent adduct between the sulfur of GSH and C7 of the substrate Pseudomonas putida (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate i.e. (E)-2'-hydroxybenzylidenepyruvate, i.e. trans-o-hydroxybenzylidenepyruvate ?

Subunits

EC Number Subunits Comment Organism
2.5.1.18 dimer
-
 Pseudomonas putida
5.99.1.4 dimer
-
 Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
2.5.1.18 2-hydroxychromene-2-carboxylic acid isomerase
-
 Pseudomonas putida
2.5.1.18 HCCA isomerase
-
 Pseudomonas putida
2.5.1.18 kappa class glutathione transferase
-
 Pseudomonas putida
2.5.1.18 kappa class GSH transferase
-
 Pseudomonas putida
5.99.1.4 2-hydroxychromene-2-carboxylic acid isomerase
-
 Pseudomonas putida
5.99.1.4 HCCA isomerase
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.18 25
-
 assay at Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.18 19
-
 trans-o-hydroxybenzylidene pyruvic acid pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
2.5.1.18 39
-
 reduced glutathione pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
2.5.1.18 47
-
 2-hydroxychromene-2-carboxylic acid pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
5.99.1.4 19
-
 (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate pH 7.0, 25°C Pseudomonas putida
5.99.1.4 47
-
 2-hydroxy-2H-chromene-2-carboxylate pH 7.0, 25°C Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.18 7
-
 assay at Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.18 GSH binding structure, overview Pseudomonas putida
5.99.1.4 glutathione the dimeric protein binds one molecule of GSH very tightly and a second molecule of GSH with much lower affinity. The enzyme is unstable in the absence of GSH. The turnover number in the forward direction greatly exceeds off rates for GSH, suggesting that GSH acts as a tightly bound cofactor in the reaction. Km: 0.017 mM Pseudomonas putida

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.5.1.18 0.136
-
 trans-o-hydroxybenzylidene pyruvic acid pH 7.0, 25°C, recombinant enzyme Pseudomonas putida
5.99.1.4 0.136
-
 (3E)-4-(2-hydroxyphenyl)-2-oxobut-3-enoate pH 7.0, 25°C Pseudomonas putida