EC Number | Cloned (Comment) | Organism |
---|---|---|
3.7.1.3 | expression of the recombinant enzyme in Escherichia coli strain DH5alpha | Pseudomonas fluorescens |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.7.1.3 | Y226F | site-directed mutagenesis, the mutant has approximately 3000fold lower activity than wild-type, and does not show the pKa values of 6.8 on kcat and 6.5 and 8.8 on kcat/Km seen for the wild-type enzyme | Pseudomonas fluorescens |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.7.1.3 | (alphaS,4S)-dihydrokynurenine | the (alphaS,4R) diastereomer is a slow substrate for a retro-aldol reaction, and the (alphaS,4S) isomer is a potent competitive inhibitor, proposed mechanism for reaction of dihydrokynurenine | Pseudomonas fluorescens | |
3.7.1.3 | 3-hydroxyhippuric acid | a competitive inhibitor | Pseudomonas fluorescens | |
3.7.1.3 | 5-bromodihydrokynurenine | a potent transition-state analogue inhibitor | Pseudomonas fluorescens | |
3.7.1.3 | S-(2-aminophenyl)-L-cysteine dioxide | - |
Pseudomonas fluorescens |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.7.1.3 | L-kynurenine + H2O | Pseudomonas fluorescens | - |
anthranilate + L-alanine | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.7.1.3 | Pseudomonas fluorescens | P83788 | gene kynU | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.7.1.3 | recombinant enzyme from Escherichia coli strain DH5alpha | Pseudomonas fluorescens |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
3.7.1.3 | L-kynurenine + H2O = anthranilate + L-alanine | the proposed mechanism of the retro-Claisen reaction requires extensive acid/base catalysis, the phosphate of pyridoxal 5'-phosphate is an acid/base catalyst in the mechanism of Pseudomonas fluorescens kynureninase, Tyr226 hydrogen bonds to the phosphate of the pyridoxal 5'-phosphate cofactor. Tyr226 mediates proton transfer between the substrate and the phosphate, which accelerates formation of external aldimine and gem-diol intermediates, catalytic reaction mechanism via several intermediates, e.g. external aldimine, quinonoid, ketimine, Gem-diolate, and enamine, overview | Pseudomonas fluorescens |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.7.1.3 | (alphaS,4R)-dihydrokynurenine + H2O | the (alphaS,4R) diastereomer is a slow substrate for a retro-aldol reaction, and the (alphaS,4S) isomer is a potent competitive inhibitor, proposed mechanism for reaction of dihydrokynurenine | Pseudomonas fluorescens | ? | - |
? | |
3.7.1.3 | L-kynurenine + H2O | - |
Pseudomonas fluorescens | anthranilate + L-alanine | - |
? | |
3.7.1.3 | L-kynurenine + H2O | the enzyme catalyzes the hydrolytic Cbeta-Cgamma cleavage of L-kynurenine to alanine and anthranilic acid catalyzing a retro-Claisen reaction | Pseudomonas fluorescens | anthranilate + L-alanine | - |
? | |
3.7.1.3 | O-benzoyl-L-serine + H2O | O-benzoyl-L-serine is an alternate substrate for kynureninase, which undergoes a normal beta-elimination reaction, proposed mechanism for reaction of O-benzoyl-L-serine, overview | Pseudomonas fluorescens | benzoate + pyruvate + ammonium | - |
? |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.7.1.3 | 37 | - |
assay at | Pseudomonas fluorescens |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.7.1.3 | 0.0058 | - |
L-kynurenine | enzyme mutant Y226F, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 0.1 | - |
O-benzoyl-L-serine | enzyme mutant Y226F, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 2.5 | - |
O-benzoyl-L-serine | wild-type enzyme, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 16 | - |
L-kynurenine | wild-type enzyme, pH 8.0, 37°C | Pseudomonas fluorescens |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.7.1.3 | 8 | - |
assay at | Pseudomonas fluorescens |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
3.7.1.3 | pyridoxal 5'-phosphate | the phosphate of pyridoxal 5'-phosphate is an acid/base catalyst in the mechanism of kynureninase. Tyr226 hydrogen bonds to the phosphate of the pyridoxal 5'-phosphate cofactor | Pseudomonas fluorescens |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.7.1.3 | 0.00003 | - |
S-(2-aminophenyl)-L-cysteine dioxide | wild-type enzyme, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 0.00092 | - |
S-(2-aminophenyl)-L-cysteine dioxide | enzyme mutant Y226F, pH 8.0, 37°C | Pseudomonas fluorescens |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.7.1.3 | 0.0041 | - |
O-benzoyl-L-serine | enzyme mutant Y226F, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 0.53 | - |
L-kynurenine | enzyme mutant Y226F, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 11 | - |
O-benzoyl-L-serine | wild-type enzyme, pH 8.0, 37°C | Pseudomonas fluorescens | |
3.7.1.3 | 200 | - |
L-kynurenine | wild-type enzyme, pH 8.0, 37°C | Pseudomonas fluorescens |