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 | 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 | Comment | Organism | Structure |
---|---|---|---|
L-phenylalanine | feedback inhibition | Escherichia coli |
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
prephenate | Escherichia coli | involved in biosynthesis of L-phenylalanine | phenylpyruvate + H2O + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
bifunctional enzyme | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type prephenate dehydratase domain and the mutant enzymes | Escherichia coli |
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 |
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 | 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 | Comment | Organism |
---|---|---|
More | the enzyme consists of 3 domains: a chorismate mutase domain, a prephenate dehydratase domain, and a regulatory domain | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
P-protein | - |
Escherichia coli |
P-protein dehydratase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia coli |