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Literature summary extracted from

  • Prieto, M.A.; Garcia, J.L.
    Molecular characterization of 4-hydroxyphenylacetate 3-hydroxylase of Escherichia coli. A two-protein component enzyme (1994), J. Biol. Chem., 269, 22823-22829.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.36 overexpression in Escherichia coli Escherichia coli W
1.14.14.9 expressed in Escherichia coli DH1 Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.5.1.36 18679
-
 2 * 18679, calculated from sequence Escherichia coli W
1.14.14.9 18679
-
 x * 18679, SDS-PAGE, enzyme is needed for activity Escherichia coli
1.14.14.9 58781
-
 2 * 58781, homodimer, encodes the hydroxylase Escherichia coli
1.14.14.9 120000
-
 gel filtration Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.14.9 4-hydroxyphenylacetate + NADH + O2 Escherichia coli inducible chromosomally encoded meta-cleavage pathway of aromatic degradation 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.36 Escherichia coli W Q57501
-
-
1.14.14.9 Escherichia coli
-
 strain W, ATCC 11105 derived from ATCC 9637
-
1.14.14.9 no activity in Escherichia coli K-12
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.9 affinity chromatography, gel filtration Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.14.9 4-hydroxyphenylacetate + FADH2 + O2 = 3,4-dihydroxyphenylacetate + FAD + H2O two enzymes suggested to be required for hydroxylase activity, member of new family of two-component aromatic hydroxylases. The hydroxylase, a flavoprotein, is encoded by hpaB and the coupling protein by hpaC gene Escherichia coli
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.14.9 additional information
-
 120000 Da enzyme encoded by hpaB shows low hydroxylase activity which is increased in the presence of FAD and the coupling protein HpaC Escherichia coli

Storage Stability

EC Number Storage Stability Organism
1.14.14.9 4°C, 20 mM phosphate buffer, pH 8.0, concentrated with polyethylene glycol 20000 Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.9 4-hydroxyphenylacetate + NADH + O2
-
 Escherichia coli 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
 ?
1.14.14.9 4-hydroxyphenylacetate + NADH + O2 inducible chromosomally encoded meta-cleavage pathway of aromatic degradation Escherichia coli 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
 ?
1.14.14.9 phenol + ?
-
 Escherichia coli catechol + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.5.1.36 dimer 2 * 18679, calculated from sequence Escherichia coli W
1.14.14.9 ? x * 18679, SDS-PAGE, enzyme is needed for activity Escherichia coli
1.14.14.9 dimer 2 * 58781, homodimer, encodes the hydroxylase Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
1.5.1.36 HpaC
-
 Escherichia coli W

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.9 30
-
 assay at Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.9 8
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.9 FAD as redox chromophore Escherichia coli
1.14.14.9 NADH
-
 Escherichia coli