Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.2.1.51 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
Search for more literature for 4.2.1.51

Cloned(Commentary)

Cloned (Comment) Organism
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

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

Inhibitors

Inhibitors Comment Organism Structure
L-phenylalanine feedback inhibition Escherichia coli

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
 bifunctional enzyme
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
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]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
P-protein
-
 Escherichia coli
P-protein dehydratase
-
 Escherichia coli

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

Ki Value [mM]

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