Protein Variants | Comment | Organism |
---|---|---|
S51A | site-directed mutagenesis, the mutation leads to a decrease of the kcat/Km parameter for reactions with L-tyrosine and 3-fluoro-L-tyrosine by three orders of magnitude, compared to the wild-type enzyme, phenotype, overview. Influence of replacement of Ser51 by Ala on the kinetic parameters of TPL reactions with inhibitory L-phenylalanine and L-methionine, kinetics and structures, overview | Citrobacter freundii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-methionine | inhibition of alpha,beta-elimination reaction | Citrobacter freundii | |
L-phenylalanine | inhibition of alpha,beta-elimination reaction | Citrobacter freundii | |
additional information | The mutant S51A also shows intermediate formation with L-Met and L-Phe. Influence of replacement of Ser51 by Ala on the kinetic parameters of TPL reactions with L-phenylalanine and L-methionine, kinetics and structures, overview | Citrobacter freundii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics | Citrobacter freundii | |
0.1 | - |
3-fluoro-L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
0.12 | - |
S-Benzyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
0.2 | - |
L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
0.21 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
0.8 | - |
beta-chloro-L-alanine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
1.3 | - |
L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
1.3 | - |
3-fluoro-L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
1.57 | - |
S-ethyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
2.12 | - |
beta-chloro-L-alanine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
2.2 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
2.32 | - |
S-methyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
4.1 | - |
S-Benzyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
5.4 | - |
S-ethyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
10.5 | - |
S-methyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-tyrosine + H2O | Citrobacter freundii | - |
phenol + pyruvate + NH3 | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Citrobacter freundii | P31013 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-tyrosine + H2O = phenol + pyruvate + NH3 | mechanism of alpha,beta-elimination of L-tyrosine catalyzed by enzyme TPL | Citrobacter freundii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-fluoro-L-tyrosine + H2O | - |
Citrobacter freundii | 3-fluorophenol + pyruvate + NH3 | - |
r | |
beta-chloro-L-alanine + H2O | - |
Citrobacter freundii | Cl- + pyruvate + NH3 | - |
r | |
L-tyrosine + H2O | - |
Citrobacter freundii | phenol + pyruvate + NH3 | - |
r | |
additional information | fragment of active site structure of the quinonoid complex of wild-type enzyme TPL with L-alanine, overview | Citrobacter freundii | ? | - |
? | |
S-(2-nitrophenyl)-L-cysteine + H2O | - |
Citrobacter freundii | 2-nitrobenzenethiolate + pyruvate + NH3 | - |
r | |
S-benzyl-L-cysteine + H2O | - |
Citrobacter freundii | thiophenol + pyruvate + NH3 | - |
r | |
S-ethyl-L-cysteine + H2O | - |
Citrobacter freundii | ethanethiol + pyruvate + NH3 | - |
r | |
S-methyl-L-cysteine + H2O | - |
Citrobacter freundii | methanethiol + pyruvate + NH3 | - |
r |
Synonyms | Comment | Organism |
---|---|---|
TPL | - |
Citrobacter freundii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Citrobacter freundii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
3-fluoro-L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.015 | - |
S-Benzyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.03 | - |
L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.044 | - |
S-methyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.083 | - |
S-ethyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.15 | - |
beta-chloro-L-alanine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.63 | - |
S-Benzyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
1.13 | - |
S-methyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
1.4 | - |
3-fluoro-L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
3.5 | - |
L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
5.4 | - |
S-ethyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
6 | - |
beta-chloro-L-alanine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
8.7 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
9.65 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Citrobacter freundii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on | Citrobacter freundii |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | rapid scanning stopped-flow kinetic experiments, kinetics | Citrobacter freundii |
General Information | Comment | Organism |
---|---|---|
additional information | the Ser51 residue interacts with the side chain amino group of Lys257 at the stage of C-alpha-proton abstraction. This interaction ensures the correct orientation of the side chain of Lys257 accepting the C-alpha-proton of the external aldimine and stabilizes its ammonium form. Ser51 takes part in formation of a chain of hydrogen bonds which is necessary to perform the transfer of the Calpha-proton to the C-4'-position of the leaving phenol group in the reaction with the natural substrate. Ser51 is necessary for efficient C-alpha-proton abstraction and its following transfer to the 4'-position of the leaving phenol group in the alpha,beta-elimination reaction of L-tyrosine | Citrobacter freundii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0037 | - |
S-Benzyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.0042 | - |
S-methyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.0092 | - |
3-fluoro-L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.015 | - |
S-ethyl-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.022 | - |
L-tyrosine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.19 | - |
beta-chloro-L-alanine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
0.49 | - |
S-methyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
2.84 | - |
beta-chloro-L-alanine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
3.43 | - |
S-ethyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
3.9 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant mutant S51A, pH 8.0, 30°C | Citrobacter freundii | |
5.26 | - |
S-Benzyl-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
14 | - |
3-fluoro-L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
17.5 | - |
L-tyrosine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii | |
46 | - |
S-(2-nitrophenyl)-L-cysteine | recombinant wild-type enzyme, pH 8.0, 30°C | Citrobacter freundii |