Literature summary for 1.13.12.9 extracted from

Ida, K.; Kurabayashi, M.; Suguro, M.; Hiruma, Y.; Hikima, T.; Yamomoto, M.; Suzuki, H.
Structural basis of proteolytic activation of L-phenylalanine oxidase from Pseudomonas sp. P-501 (2008), J. Biol. Chem., 283, 16584-16590.

Search for more literature for 1.13.12.9

Activating Compound

Activating Compound	Comment	Organism	Structure
Pronase	-	Pseudomonas sp.
Trypsin	-	Pseudomonas sp.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the proenzyme in Escherichia coli, site-directed mutation introduced on the wild-type proenzyme gene in the plasmid pPAO+15, expression of proenzyme mutants in Escherichia coli BL21(DE3)	Pseudomonas sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals of noncatalytic proenzyme, activated enzyme, and enzyme o-amino benzoate complex, purified protein in 10 mM Tris-HCl (pH 8.0), concentrated, mixed with 0.1 M HEPES (pH 7.5) and 1.7 M ammonium sulfate (for the proenzyme) or 1.0 M ammonium sulfate (for the active enzyme), 30% glycerol in buffer as cryoprotectant, the enzyme amino benzoate complex is prepared by a soaking method with the cryoprotectant	Pseudomonas sp.

Crystallization (Comment)

Organism

crystals of noncatalytic proenzyme, activated enzyme, and enzyme o-amino benzoate complex, purified protein in 10 mM Tris-HCl (pH 8.0), concentrated, mixed with 0.1 M HEPES (pH 7.5) and 1.7 M ammonium sulfate (for the proenzyme) or 1.0 M ammonium sulfate (for the active enzyme), 30% glycerol in buffer as cryoprotectant, the enzyme amino benzoate complex is prepared by a soaking method with the cryoprotectant

Pseudomonas sp.

Protein Variants

Protein Variants	Comment	Organism
K478A	catalytic activity 1/200 of the wild-type enzyme	Pseudomonas sp.
M142A	kcat is 1/10 of the wild-type enzyme, Km is elevated, affinity for oxygen seems decreased	Pseudomonas sp.

Inhibitors

Inhibitors	Comment	Organism	Structure
2-amino benzoate	-	Pseudomonas sp.

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0159	-	L-phenylalanine	active enzyme comparable to native enzyme, 20 mM Tris-HCl, pH 8.0, 25°C	Pseudomonas sp.
3.4	-	L-phenylalanine	M142A mutant (elevated compared to 1.8 mM for wild-type), 20 mM Tris-HCl, pH 8.0, 25°C	Pseudomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-phenylalanine + O2	Pseudomonas sp.	oxidative deamination and oxygenative decarboxylation of L-phenylalanine, L-tyrosine, L-tryptophan, and L-methionine	2-phenylacetamide + CO2 + H2O	-	?
L-phenylalanine + O2	Pseudomonas sp. P-501	oxidative deamination and oxygenative decarboxylation of L-phenylalanine, L-tyrosine, L-tryptophan, and L-methionine	2-phenylacetamide + CO2 + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas sp.	-	P-501	-
Pseudomonas sp. P-501	-	P-501	-

Purification (Commentary)

Purification (Comment)	Organism
cell disruption by sonication, centrifugation, supernatant applied to HisTrap HP column equilibrated with 20 mM Tris-HCl (pH 8.0) containing 500 mM NaCl, elution with linear gradient of imidazole from 0-0.5 M, ammonium sulfate added to eluted fractions, centrifugation of precipitate, solution in 20 mM Tris-HCl (pH 8.0) containing 300 mM ammonium sulfate, several chromatographic steps using HiTrap Phenyl FF, Resource Q, and HiLoad 16/60 Superdex 200 prep grade, activated enzyme purified by HiTrap Q HP and gel filtration	Pseudomonas sp.

Purification (Comment)

Organism

cell disruption by sonication, centrifugation, supernatant applied to HisTrap HP column equilibrated with 20 mM Tris-HCl (pH 8.0) containing 500 mM NaCl, elution with linear gradient of imidazole from 0-0.5 M, ammonium sulfate added to eluted fractions, centrifugation of precipitate, solution in 20 mM Tris-HCl (pH 8.0) containing 300 mM ammonium sulfate, several chromatographic steps using HiTrap Phenyl FF, Resource Q, and HiLoad 16/60 Superdex 200 prep grade, activated enzyme purified by HiTrap Q HP and gel filtration

Pseudomonas sp.

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-phenylalanine + O2	oxidative deamination and oxygenative decarboxylation of L-phenylalanine, L-tyrosine, L-tryptophan, and L-methionine	Pseudomonas sp.	2-phenylacetamide + CO2 + H2O	-	?
L-phenylalanine + O2	oxidative deamination and oxygenative decarboxylation of L-phenylalanine, L-tyrosine, L-tryptophan, and L-methionine	Pseudomonas sp. P-501	2-phenylacetamide + CO2 + H2O	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * x, N-terminal 14 residue prosequence, alpha subunit, dipeptide, beta subunit	Pseudomonas sp.

Synonyms

Synonyms	Comment	Organism
L-phenylalanine oxidase	-	Pseudomonas sp.
PAOpt	activated enzyme	Pseudomonas sp.
proPAO	noncatalytic proenzyme, activated by proteolytic cleavage	Pseudomonas sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
115	-	L-phenylalanine	active enzyme comparable to native enzyme, 20 mM Tris-HCl, pH 8.0, 25°C	Pseudomonas sp.

Cofactor

Cofactor	Comment	Organism	Structure
flavin adenine dinucleotide	FAD	Pseudomonas sp.

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.87	-	2-amino benzoate	20 mM Tris-HCl, pH 8.0, 25°C	Pseudomonas sp.