EC Number | Cloned (Comment) | Organism |
---|---|---|
1.13.11.27 | expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Streptomyces avermitilis |
1.13.11.46 | expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Amycolatopsis orientalis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.13.11.27 | N245I | site-directed mutagenesis, the mutant enzyme produces 13% homogentisate and 87% 4-hydroxyphenylacetate, which differs from the wild-type activity | Streptomyces avermitilis |
1.13.11.27 | 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 |
1.13.11.27 | 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 |
1.13.11.27 | 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 |
1.13.11.46 | 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 |
1.13.11.46 | 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 |
1.13.11.46 | T214P | site-directed mutagenesis, mutant shows decreased catalytic efficiency compared to the wild-type enzyme | Amycolatopsis orientalis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.27 | 0.0087 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant P243T | Streptomyces avermitilis | |
1.13.11.27 | 0.015 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant N245Q | Streptomyces avermitilis | |
1.13.11.27 | 0.027 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme and mutant N245I | Streptomyces avermitilis | |
1.13.11.27 | 0.047 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S230A | Streptomyces avermitilis | |
1.13.11.46 | 0.0143 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant I216N | Amycolatopsis orientalis | |
1.13.11.46 | 0.0205 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant T214P | Amycolatopsis orientalis | |
1.13.11.46 | 0.028 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S201A | Amycolatopsis orientalis | |
1.13.11.46 | 0.0476 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme | Amycolatopsis orientalis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.13.11.46 | Fe2+ | a Fe(II)-dependent dioxygenase | Amycolatopsis orientalis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.27 | 4-hydroxyphenylpyruvate + O2 | Streptomyces avermitilis | - |
homogentisate + CO2 + 4-hydroxyphenylacetate | the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate | ? | |
1.13.11.46 | 4-hydroxyphenylpyruvate + O2 | Amycolatopsis orientalis | - |
4-hydroxymandelate + CO2 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.13.11.27 | Streptomyces avermitilis | - |
- |
- |
1.13.11.46 | Amycolatopsis orientalis | - |
gene AOHMS | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.13.11.27 | recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration | Streptomyces avermitilis |
1.13.11.46 | recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration | Amycolatopsis orientalis |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.13.11.27 | 4-hydroxyphenylpyruvate + O2 = homogentisate + CO2 | reaction mechanism, overview | Streptomyces avermitilis | |
1.13.11.46 | 4-hydroxyphenylpyruvate + O2 = (S)-4-hydroxymandelate + CO2 | reaction mechanism, overview | Amycolatopsis orientalis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.13.11.27 | 4-hydroxyphenylpyruvate + O2 | - |
Streptomyces avermitilis | homogentisate + CO2 + 4-hydroxyphenylacetate | the wild-type enzyme produces 85% homogentisate and 15% 4-hydroxyphenylacetate | ? | |
1.13.11.46 | 4-hydroxyphenylpyruvate + O2 | - |
Amycolatopsis orientalis | 4-hydroxymandelate + CO2 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.13.11.27 | HPPD | - |
Streptomyces avermitilis |
1.13.11.46 | hydroxymandelate synthase | - |
Amycolatopsis orientalis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.13.11.27 | 25 | - |
assay at | Streptomyces avermitilis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.27 | 0.03 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant N245Q | Streptomyces avermitilis | |
1.13.11.27 | 0.64 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant N245I | Streptomyces avermitilis | |
1.13.11.27 | 2.1 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant P243T | Streptomyces avermitilis | |
1.13.11.27 | 2.8 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S230A | Streptomyces avermitilis | |
1.13.11.27 | 6.8 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme | Streptomyces avermitilis | |
1.13.11.46 | 1.2 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant T214P | Amycolatopsis orientalis | |
1.13.11.46 | 1.4 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant I216N | Amycolatopsis orientalis | |
1.13.11.46 | 3 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S201A | Amycolatopsis orientalis | |
1.13.11.46 | 3.7 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme | Amycolatopsis orientalis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.13.11.27 | 7 | - |
assay at | Streptomyces avermitilis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.13.11.46 | 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 |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.13.11.27 | 2.4 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant N245Q | Streptomyces avermitilis | |
1.13.11.27 | 24 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant N245I | Streptomyces avermitilis | |
1.13.11.27 | 60 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S230A | Streptomyces avermitilis | |
1.13.11.27 | 241 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant P243T | Streptomyces avermitilis | |
1.13.11.27 | 254 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme | Streptomyces avermitilis | |
1.13.11.46 | 57.5 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant T214P | Amycolatopsis orientalis | |
1.13.11.46 | 77 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, wild-type enzyme | Amycolatopsis orientalis | |
1.13.11.46 | 98.5 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant I216N | Amycolatopsis orientalis | |
1.13.11.46 | 108 | - |
4-hydroxyphenylpyruvate | pH 7.0, 25°C, mutant S201A | Amycolatopsis orientalis |