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Literature summary for 1.14.14.9 extracted from

  • Hawumba, J.F.; Broezel, V.S.; Theron, J.
    Cloning and characterization of a 4-hydroxyphenylacetate 3-hydroxylase from the thermophile Geobacillus sp. PA-9 (2007), Curr. Microbiol., 55, 480-484.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene hpaH, DNA and amino acid sequence determination and analysis, sequence comparisons, expression in enzyme-deficient Escherichia coli strain DH5alpha Geobacillus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56000
-
x * 56269, sequence calculation, x * 56000, recombinant enzyme, SDS-PAGE Geobacillus sp.
56269
-
x * 56269, sequence calculation, x * 56000, recombinant enzyme, SDS-PAGE Geobacillus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-hydroxyphenylacetate + NADH + H+ + O2 Geobacillus sp.
-
3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?
4-hydroxyphenylacetate + NADH + H+ + O2 Geobacillus sp. PA-9
-
3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus sp. Q4L1M7 gene hpaH
-
Geobacillus sp. PA-9 Q4L1M7 gene hpaH
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain DH5alpha by affinity chromatography on a 4-hydroxyphenylacetate-coupled amino-agarose column to near homogeneity Geobacillus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxyphenylacetate + NADH + H+ + O2
-
Geobacillus sp. 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?
4-hydroxyphenylacetate + NADH + H+ + O2 compared with intracellular cell extracts, the purified recombinant HpaH protein does not display detectable activity toward the 4-hydroxyphenylacetate substrate under the assay conditions used, in contrast to the cell extract, overview Geobacillus sp. 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?
4-hydroxyphenylacetate + NADH + H+ + O2
-
Geobacillus sp. PA-9 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?
4-hydroxyphenylacetate + NADH + H+ + O2 compared with intracellular cell extracts, the purified recombinant HpaH protein does not display detectable activity toward the 4-hydroxyphenylacetate substrate under the assay conditions used, in contrast to the cell extract, overview Geobacillus sp. PA-9 3,4-dihydroxyphenylacetate + NAD+ + H2O
-
?

Subunits

Subunits Comment Organism
? x * 56269, sequence calculation, x * 56000, recombinant enzyme, SDS-PAGE Geobacillus sp.

Synonyms

Synonyms Comment Organism
4-HPA hydroxylase
-
Geobacillus sp.
HPAH
-
Geobacillus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
assay at Geobacillus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
assay at Geobacillus sp.

Cofactor

Cofactor Comment Organism Structure
NADH
-
Geobacillus sp.