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Literature summary for 3.7.1.25 extracted from

  • Duggleby, C.; Williams, P.
    Purification and some properties of the 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase (2-hydroxymuconic semialdehyde hydrolase) encoded by the TOL plasmid pWW0 from Pseudomonas putida mt-2 (1986), J. Gen. Microbiol., 132, 717-726 .
No PubMed abstract available

Activating Compound

Activating Compound Comment Organism Structure
additional information neither EDTA, 2,2'-bipyridyl nor diethyldithiocarbamate have any effect upon enzyme activity. The activity is unaffected by reduced glutathione, 2-mercaptoethanol or L-cysteine (all at 1 mM) Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
Ag+ Ag+ concentrations in the range 0.001-0.01 mM lower the activity to less than 50% of its uninhibited value Pseudomonas putida
Cu2+ Cu2+ concentrations in the range 0.001-0.01 mM lower the activity to less than 50% of its uninhibited value Pseudomonas putida
iodoacetamide 2 mM iodoacetate inhibits the enzyme activity to 50% or less of its uninhibited value after 15 min incubation with the enzyme at 0°C Pseudomonas putida
p-chloromercuribenzoate 0.2 mM p-chloromercuribenzoate inhibits the enzyme activity to 50% or less of its uninhibited value immediately after incubation with the enzyme at 0°C Pseudomonas putida
sodium arsenite 0.2 mM sodium arsenite inhibits the enzyme activity to 50% or less of its uninhibited value after 15 min incubation with the enzyme at 0°C Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.036
-
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate at pH 7.5 and 25°C Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
65000
-
gel filtration Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O Pseudomonas putida
-
(2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O Pseudomonas putida ATCC 33015
-
(2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
mt-2
-
Pseudomonas putida ATCC 33015
-
mt-2
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, CM-cellulose column chromatography, DEAE-cellulose column chromatography, Sephadex G-100 gel filtration, and QAE-Sephadex A-50 gel filtration Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.58
-
cell-free extract, at pH 7.5 and 25°C Pseudomonas putida
48
-
after 82.8fold purification, at pH 7.5 and 25°C Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
Pseudomonas putida (2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O 100% activity Pseudomonas putida (2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
Pseudomonas putida ATCC 33015 (2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?
(2Z,4E)-2-hydroxy-6-oxohepta-2,4-dienoate + H2O 100% activity Pseudomonas putida ATCC 33015 (2Z)-2-hydroxypenta-2,4-dienoate + acetate
-
?

Subunits

Subunits Comment Organism
homodimer 31000-33000, SDS-PAGE Pseudomonas putida

Synonyms

Synonyms Comment Organism
2-hydroxymuconic semialdehyde hydrolase
-
Pseudomonas putida
HOD hydrolase
-
Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Pseudomonas putida

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 9 the enzyme is fairly stable to pH values between 5.5 to 9.0, with maximum stability at 7.0 to 8.0. Loss of activity is rapid at pH values below 5.0 and above 9.5 Pseudomonas putida

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas putida isoelectric focusing
-
4.7