Literature summary for 3.7.1.3 extracted from

Momany, C.; Levdikov, V.; Blagova, L.; Lima, S.; Phillips, R.S.
Three-dimensional structure of kynureninase from Pseudomonas fluorescens (2004), Biochemistry, 43, 1193-1203.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Pseudomonas fluorescens
subcloned into pSapKO-WT and transformed into Escherichia coli B834(DE3)	Pseudomonas fluorescens

Crystallization (Commentary)

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

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

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Pseudomonas fluorescens	P83788	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
no modification	-	Pseudomonas fluorescens

Purification (Commentary)

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

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-kynurenine + H2O	L-tryptophan metabolism	Pseudomonas fluorescens	anthranilate + L-alanine	-	?

Subunits

Subunits	Comment	Organism
dimer	-	Pseudomonas fluorescens

Synonyms

Synonyms	Comment	Organism
L-kynurenine hydrolase	-	Pseudomonas fluorescens

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	-	Pseudomonas fluorescens