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Literature summary for 1.13.11.46 extracted from

  • Shah, D.D.; Conrad, J.A.; Heinz, B.; Brownlee, J.M.; Moran, G.R.
    Evidence for the mechanism of hydroxylation by 4-hydroxyphenylpyruvate dioxygenase and hydroxymandelate synthase from intermediate partitioning in active site variants (2011), Biochemistry, 50, 7694-7704.
    View publication on PubMed
Search for more literature for 1.13.11.46

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Amycolatopsis orientalis

Protein Variants

Protein Variants Comment Organism
I216N site-directed mutagenesis, the mutant shows increased catalytic efficiency compared to the wild-type enzyme, the mutant produces 90% 4-hydroxmandelate and 10% 4-hydroxyphenylacetate from 4-hydroxyphenylpyruvate, which differs from the wild-type, that does not produce any 4-hydroxyphenylacetate Amycolatopsis orientalis
S201A site-directed mutagenesis, the mutant shows increased catalytic efficiency compared to the wild-type enzyme, the mutant produces 88% 4-hydroxmandelate and 12% 4-hydroxyphenylacetate from 4-hydroxyphenylpyruvate, which differs from the wild-type, that does not produce any 4-hydroxyphenylacetate Amycolatopsis orientalis
T214P site-directed mutagenesis, mutant shows decreased catalytic efficiency compared to the wild-type enzyme Amycolatopsis orientalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0143
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant I216N Amycolatopsis orientalis
0.0205
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant T214P Amycolatopsis orientalis
0.028
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant S201A Amycolatopsis orientalis
0.0476
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, wild-type enzyme Amycolatopsis orientalis

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ a Fe(II)-dependent dioxygenase Amycolatopsis orientalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-hydroxyphenylpyruvate + O2 Amycolatopsis orientalis
-
 4-hydroxymandelate + CO2
-
 ?

Organism

Organism UniProt Comment Textmining
Amycolatopsis orientalis
-
 gene AOHMS
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration Amycolatopsis orientalis

Reaction

Reaction Comment Organism Reaction ID
4-hydroxyphenylpyruvate + O2 = (S)-4-hydroxymandelate + CO2 reaction mechanism, overview Amycolatopsis orientalis
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxyphenylpyruvate + O2
-
 Amycolatopsis orientalis 4-hydroxymandelate + CO2
-
 ?

Synonyms

Synonyms Comment Organism
hydroxymandelate synthase
-
 Amycolatopsis orientalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant T214P Amycolatopsis orientalis
1.4
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant I216N Amycolatopsis orientalis
3
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant S201A Amycolatopsis orientalis
3.7
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, wild-type enzyme Amycolatopsis orientalis

General Information

General Information Comment Organism
evolution 4-hydroxyphenylpyruvate dioxygenase, HPPD, EC 1.13.11.27, and hydroxymandelate synthase catalyze similar reactions using the same substrates, 4-hydroxyphenylpyruvate and dioxygen. Initially, both enzymes reduce and activate dioxygen in order to decarboxylate 4-hydroxyphenylpyruvate, yielding 4-hydroxyphenylacetate, CO2, and an activated oxo intermediate. Both enzymes then hydroxylate 4-hydroxyphenylacetate but do so in different positions Amycolatopsis orientalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
57.5
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant T214P Amycolatopsis orientalis
77
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, wild-type enzyme Amycolatopsis orientalis
98.5
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant I216N Amycolatopsis orientalis
108
-
 4-hydroxyphenylpyruvate pH 7.0, 25°C, mutant S201A Amycolatopsis orientalis