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Literature summary extracted from

  • Burks, E.A.; Yan, W.; Johnson, W.H.; Li, W.; Schroeder, G.K.; Min, C.; Gerratana, B.; Zhang, Y.; Whitman, C.P.
    Kinetic, crystallographic, and mechanistic characterization of TomN: elucidation of a function for a 4-oxalocrotonate tautomerase homologue in the tomaymycin biosynthetic pathway (2011), Biochemistry, 50, 7600-7611.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.3.2.6 expressed in Escherichia coli BL21-Gold(DE3) cells Pseudomonas putida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
5.3.2.6 sitting drop vapor diffusion method, using 100 mM magnesium acetate/100 mM sodium acetate, 5%-21% (w/v) PEG 8000, pH 4.5, and 100 mM calcium acetate/100 mM sodium acetate, 1%-13% (w/v) PEG 4000, pH 4.5 Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
5.3.2.6 P1A the mutation results in 430fold decreases in kcat/Km compared to the wild type enzyme Pseudomonas putida
5.3.2.6 R11A the mutation results in 280fold decreases in kcat/Km compared to the wild type enzyme Pseudomonas putida
5.3.2.6 R39A the mutation results in 8000fold decreases in kcat/Km compared to the wild type enzyme Pseudomonas putida
5.3.2.6 R61A the mutation results in 8fold decreases in kcat/Km compared to the wild type enzyme Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.3.2.6 2-Oxo-3-pentynoate
-
 Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.3.2.6 0.18
-
 2-hydroxymuconate mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 0.334
-
 (2E)-2-hydroxy-3-phenylprop-2-enoate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 0.44
-
 2-hydroxymuconate mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 0.512
-
 2-hydroxymuconate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 0.78
-
 2-hydroxymuconate mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 1.05
-
 2-hydroxymuconate mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.3.2.6 (2Z,4E)-2-hydroxyhexa-2,4-dienedioate Pseudomonas putida
-
 (3E)-2-oxohex-3-enedioate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.3.2.6 Pseudomonas putida
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.3.2.6 phenyl-Sepharose column chromatography and DEAE-Sepharose column chromatography Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.2.6 (2E)-2-hydroxy-3-phenylprop-2-enoate
-
 Pseudomonas putida 2-oxo-3-phenylpropanoate
-
 ?
5.3.2.6 (2Z,4E)-2-hydroxyhexa-2,4-dienedioate
-
 Pseudomonas putida (3E)-2-oxohex-3-enedioate
-
 r
5.3.2.6 (3E)-6-oxohept-3-enedioate
-
 Pseudomonas putida 2-hydroxy-2,4-heptadiene-1,7-dioate
-
 ?
5.3.2.6 2-hydroxy-2,4-heptadiene-1,7-dioate
-
 Pseudomonas putida (3E)-2-oxohept-3-enedioate
-
 ?
5.3.2.6 2-hydroxy-2,4-pentadienoate
-
 Pseudomonas putida (3E)-2-oxopent-3-enoate
-
 ?
5.3.2.6 2-hydroxymuconate
-
 Pseudomonas putida 2-oxo-3-hexenedioate
-
 r
5.3.2.6 2-oxopent-4-enoate
-
 Pseudomonas putida 2-hydroxy-2,4-pentadienoate
-
 ?
5.3.2.6 4-vinyl-2,3-dihydropyrrole-2-carboxylic acid
-
 Pseudomonas putida 4-ethylidene-3,4-dihydropyrrole-2-carboxylic acid
-
 r

Synonyms

EC Number Synonyms Comment Organism
5.3.2.6 4-OT
-
 Pseudomonas putida
5.3.2.6 4-oxalocrotonate tautomerase
-
 Pseudomonas putida
5.3.2.6 TomN
-
 Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3.2.6 0.2
-
 2-hydroxymuconate mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 1.5
-
 2-hydroxymuconate mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 14
-
 2-hydroxymuconate mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 16
-
 (2E)-2-hydroxy-3-phenylprop-2-enoate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 360
-
 2-hydroxymuconate mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 1850
-
 2-hydroxymuconate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.3.2.6 0.45
-
 2-hydroxymuconate mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 8.3
-
 2-hydroxymuconate mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 13
-
 2-hydroxymuconate mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 48
-
 (2E)-2-hydroxy-3-phenylprop-2-enoate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 460
-
 2-hydroxymuconate mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida
5.3.2.6 3600
-
 2-hydroxymuconate wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C Pseudomonas putida