Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Pseudomonas fluorescens |
subcloned into pSapKO-WT and transformed into Escherichia coli B834(DE3) | Pseudomonas fluorescens |
Crystallization (Comment) | Organism |
---|---|
by hanging-drop vapor diffusion method at room temperature, to 1.85 A resolution, Lys-227 is the pyridoxal-5'-phosphate binding residue near the pyridoxal-5'-phosphate nitrogen, but Asp-132 is also strictly conserved and at a similar distance from the pyridinium nitrogen, Tyr-226 donates a hydrogen bond to the phosphate of pyridoxal-5'-phosphate, Trp-256 donates a hydrogen bond to the phosphate through the indole N1-hydrogen | Pseudomonas fluorescens |
performed using the hanging-drop vapor diffusion technique | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
D132A | has lower activity and binds pyridoxal 5'-phosphate weakly | Pseudomonas fluorescens |
D132A | has lower activity and weaklier pyridoxal-5'-phosphate binding than that of wild-type enzyme | Pseudomonas fluorescens |
D132E | has good activity and pyridoxal 5'-phosphate binding stronger than that of the wild-type enzyme | Pseudomonas fluorescens |
D132E | has good activity and stronger pyridoxal-5'-phosphate binding than that of wild-type enzyme | Pseudomonas fluorescens |
D201A | has very low activity, less than 0.01% of that of the wild-type enzyme, binds pyridoxal 5'-phosphate weakly | Pseudomonas fluorescens |
D201A | has very low activity and binds pyridoxal-5'-phosphate weakly, and expresses poorly, giving large amounts of insoluble inclusion bodies and very low levels of soluble protein | Pseudomonas fluorescens |
D201E | has good activity and pyridoxal 5'-phosphate binding stronger than that of the wild-type enzyme | Pseudomonas fluorescens |
D201E | has good activity and stronger pyridoxal-5'-phosphate binding than that of wild-type enzyme | Pseudomonas fluorescens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-kynurenine + H2O | Pseudomonas fluorescens | L-tryptophan metabolism | anthranilate + L-alanine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas fluorescens | P83788 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
no modification | - |
Pseudomonas fluorescens |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas fluorescens |
recombinant enzyme, by centrifugation, protamine sulfate precipitation, HiTrap Q column, omega-aminohexylagarose column, second HiTrap Q column and phenyl-Sepharose column | Pseudomonas fluorescens |
Storage Stability | Organism |
---|---|
-78ºC, 0.1 mM potassium phosphate buffer, 0.2 mM pyridoxal 5'-phosphate, pH 8.0, 1 year, stable | Pseudomonas fluorescens |
-78ºC, 5 mM potassium phosphate buffer, 0.2 mM pyridoxal 5'-phosphate, pH 8.0, 1 month, loses 4% of activity | Pseudomonas fluorescens |
4ºC, 5 mM potassium phosphate buffer, 0.2 mM pyridoxal 5'-phosphate, pH 8.0, 1 week, loses 40% of activity | Pseudomonas fluorescens |
78°C, phosphate buffer, 1 year | Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-kynurenine + H2O | L-tryptophan metabolism | Pseudomonas fluorescens | anthranilate + L-alanine | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Pseudomonas fluorescens |
Synonyms | Comment | Organism |
---|---|---|
L-kynurenine hydrolase | - |
Pseudomonas fluorescens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Pseudomonas fluorescens |