Literature summary for 1.13.11.27 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.

Search for more literature for 1.13.11.27

Cloned(Commentary)

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

Protein Variants

Protein Variants	Comment	Organism
N245I	site-directed mutagenesis, the mutant enzyme produces 13% homogentisate and 87% 4-hydroxyphenylacetate, which differs from the wild-type activity	Streptomyces avermitilis
N245Q	site-directed mutagenesis, the mutant enzyme produces 44.5% homogentisate and 52% 4-hydroxyphenylacetate, and 4% quinolacetate, which differs from the wild-type activity	Streptomyces avermitilis
P243T	site-directed mutagenesis, the mutant enzyme produces 13% homogentisate, 8.7% 4-hydroxyphenylacetate, and 78% quinolacetate, which differs from the wild-type activity	Streptomyces avermitilis
S230A	site-directed mutagenesis, the mutant enzyme produces 11.6% homogentisate, 30.6% 4-hydroxyphenylacetate, and 57.8% quinolacetate, which differs from the wild-type activity	Streptomyces avermitilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0087	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant P243T	Streptomyces avermitilis
0.015	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant N245Q	Streptomyces avermitilis
0.027	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, wild-type enzyme and mutant N245I	Streptomyces avermitilis
0.047	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant S230A	Streptomyces avermitilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
4-hydroxyphenylpyruvate + O2	Streptomyces avermitilis	-	homogentisate + CO2 + 4-hydroxyphenylacetate	the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate	?

Organism

Organism	UniProt	Comment	Textmining
Streptomyces avermitilis	-	-	-

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	Streptomyces avermitilis

Reaction

Reaction	Comment	Organism	Reaction ID
4-hydroxyphenylpyruvate + O2 = homogentisate + CO2	reaction mechanism, overview	Streptomyces avermitilis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-hydroxyphenylpyruvate + O2	-	Streptomyces avermitilis	homogentisate + CO2 + 4-hydroxyphenylacetate	the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate	?

Synonyms

Synonyms	Comment	Organism
HPPD	-	Streptomyces avermitilis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Streptomyces avermitilis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.03	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant N245Q	Streptomyces avermitilis
0.64	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant N245I	Streptomyces avermitilis
2.1	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant P243T	Streptomyces avermitilis
2.8	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant S230A	Streptomyces avermitilis
6.8	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, wild-type enzyme	Streptomyces avermitilis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Streptomyces avermitilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
2.4	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant N245Q	Streptomyces avermitilis
24	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant N245I	Streptomyces avermitilis
60	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant S230A	Streptomyces avermitilis
241	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, mutant P243T	Streptomyces avermitilis
254	-	4-hydroxyphenylpyruvate	pH 7.0, 25°C, wild-type enzyme	Streptomyces avermitilis

Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.

Release 2023.1 (January 2023)