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Literature summary extracted from

  • Zhang, S.; Wilson, D.B.; Ganem, B.
    Probing the catalytic mechanism of prephenate dehydratase by site-directed mutagenesis of the Escherichia coli P-protein dehydratase domain (2000), Biochemistry, 39, 4722-4728.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.51 expression of mutant enzymes in enzyme deficient strain NK6024, subcloning in strain DH5alpha, overexpression of the isolated His-tagged wild-type prephenate dehydratase domain in strain BL21(DE3) Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.51 C216A site-directed mutagenesis, inactive mutant Escherichia coli
4.2.1.51 C216S site-directed mutagenesis, increased activity Escherichia coli
4.2.1.51 E159A site-directed mutagenesis, 2.2fold increased activity Escherichia coli
4.2.1.51 E159A/E232A site-directed mutagenesis, 7fold increased kcat, 4.6fold decreased Km compared to the wild-type, increased activity Escherichia coli
4.2.1.51 E232A site-directed mutagenesis, 3.5fold increased activity Escherichia coli
4.2.1.51 H209A site-directed mutagenesis, highly reduced activity Escherichia coli
4.2.1.51 N160A site-directed mutagenesis, 500fold decreased activity Escherichia coli
4.2.1.51 N160D site-directed mutagenesis, highly reduced activity Escherichia coli
4.2.1.51 Q215A site-directed mutagenesis, 500fold decreased activity Escherichia coli
4.2.1.51 S208A site-directed mutagenesis, 100fold decreased activity Escherichia coli
4.2.1.51 S208C site-directed mutagenesis, 100fold decreased activity Escherichia coli
4.2.1.51 S208D site-directed mutagenesis, inactive mutant Escherichia coli
4.2.1.51 T278A site-directed mutagenesis, catalytically inactive mutant, but binds to substrate and inhibitor Escherichia coli
4.2.1.51 T278S site-directed mutagenesis, 100fold decreased activity Escherichia coli
4.2.1.51 T278V site-directed mutagenesis, catalytically inactive mutant, but binds to substrate and inhibitor Escherichia coli
4.2.1.51 W226A site-directed mutagenesis, nearly inactive mutant Escherichia coli
4.2.1.51 W226L site-directed mutagenesis, highly reduced activity Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.51 L-phenylalanine feedback inhibition Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.51 additional information
-
 additional information Km for chorismate of wild-type and mutants Escherichia coli
4.2.1.51 0.12
-
 prephenate mutant E159A/E232A, 37°C Escherichia coli
4.2.1.51 0.16
-
 prephenate mutant E232A, 37°C Escherichia coli
4.2.1.51 0.25
-
 prephenate mutant E159A, 37°C Escherichia coli
4.2.1.51 0.56
-
 prephenate native P-protein, 37°C Escherichia coli
4.2.1.51 0.57
-
 prephenate mutant T278S, 37°C Escherichia coli
4.2.1.51 0.65
-
 prephenate recombinant prephenate dehydratase domain, 37°C Escherichia coli
4.2.1.51 1.65
-
 prephenate mutant N160A, 37°C Escherichia coli
4.2.1.51 2.09
-
 prephenate mutant C216S, 37°C Escherichia coli
4.2.1.51 3
-
 prephenate above, mutant W226L, 37°C Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.51 prephenate Escherichia coli involved in biosynthesis of L-phenylalanine phenylpyruvate + H2O + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.51 Escherichia coli
-
 bifunctional enzyme
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.51 recombinant wild-type prephenate dehydratase domain and the mutant enzymes Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.1.51 prephenate = phenylpyruvate + H2O + CO2 this enzyme in the enteric bacteria also possesses chorismate mutase activity, EC 5.4.99.5, and converts chorismate into prephenate, catalytic mechanism, T278 is involved in catalysis Escherichia coli
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.1.51 additional information
-
 substrate specificity of the bifunctional enzyme, activity of mutant enzymes, overview Escherichia coli
4.2.1.51 18.2
-
 crude native cell extract Escherichia coli
4.2.1.51 37.6
-
 purified recombinant prephenate dehydratase domain Escherichia coli
4.2.1.51 40.4
-
 purified recombinant mutant E159A Escherichia coli
4.2.1.51 42.9
-
 purified recombinant mutant E232A Escherichia coli
4.2.1.51 56.9
-
 purified recombinant double mutant E159A/E232A Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.51 prephenate
-
 Escherichia coli phenylpyruvate + H2O + CO2
-
 ?
4.2.1.51 prephenate involved in biosynthesis of L-phenylalanine Escherichia coli phenylpyruvate + H2O + CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.51 More the enzyme consists of 3 domains: a chorismate mutase domain, a prephenate dehydratase domain, and a regulatory domain Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
4.2.1.51 P-protein
-
 Escherichia coli
4.2.1.51 P-protein dehydratase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.51 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.51 additional information
-
 additional information kcat for chorismate of wild-type and mutants Escherichia coli
4.2.1.51 0.22
-
 prephenate mutant N160A, 37°C Escherichia coli
4.2.1.51 0.41
-
 prephenate mutant T278S, 37°C Escherichia coli
4.2.1.51 20
-
 prephenate mutant W226L, 37°C Escherichia coli
4.2.1.51 32
-
 prephenate native P-protein, 37°C Escherichia coli
4.2.1.51 32
-
 prephenate recombinant prephenate dehydratase domain, 37°C Escherichia coli
4.2.1.51 35
-
 prephenate mutant C216S, 37°C Escherichia coli
4.2.1.51 40
-
 prephenate mutant E232A, 37°C Escherichia coli
4.2.1.51 42
-
 prephenate mutant E159A, 37°C Escherichia coli
4.2.1.51 49
-
 prephenate mutant E159A/E232A, 37°C Escherichia coli

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.2.1.51 additional information
-
 additional information
-
 Escherichia coli