Literature summary for 3.5.1.54 extracted from

Balotra, S.; Newman, J.; French, N.G.; Briggs, L.J.; Peat, T.S.; Scott, C.
Crystallization and preliminary X-ray diffraction analysis of the amidase domain of allophanate hydrolase from Pseudomonas sp. strain ADP (2014), Acta crystallogr. Sect. F, 70, 310-315.

Cloned(Commentary)

Cloned (Comment)	Organism
gene atzF, DNA and amino acid sequence determination and analysis, recombinant expression of functional His-tagged enzyme in Escherichia coli strain BL21 (lambdaDE3)	Pseudomonas sp.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified full-length wild-type enzyme and truncated mutant enzyme, trypsin-treated AtzF (in situ proteolysis) from 1 M ammonium sulfate, 1 M lithium sulfate, 0.1 M Tris-HCl, pH 8.5, AtzF467 crystals grown from 20% w/v PEG 6000, 0.1 M Na MES pH 6.5, 0.2 M calcium chloride, are used in microseeding for truncated AtzF crystal growth from 11% w/v PEG 3350, 2% Tacsimate, pH 5.0, X-ray diffraction structure determination and analysis at 2.5 A resolution	Pseudomonas sp.

Crystallization (Comment)

Organism

purified full-length wild-type enzyme and truncated mutant enzyme, trypsin-treated AtzF (in situ proteolysis) from 1 M ammonium sulfate, 1 M lithium sulfate, 0.1 M Tris-HCl, pH 8.5, AtzF467 crystals grown from 20% w/v PEG 6000, 0.1 M Na MES pH 6.5, 0.2 M calcium chloride, are used in microseeding for truncated AtzF crystal growth from 11% w/v PEG 3350, 2% Tacsimate, pH 5.0, X-ray diffraction structure determination and analysis at 2.5 A resolution

Pseudomonas sp.

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of the gene encoding C-terminally truncated AtzF mutant, AtzF467	Pseudomonas sp.

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
50000	-	6 * 50000, recombinant enzyme, SDS-PAGE	Pseudomonas sp.
360000	-	gel filtration	Pseudomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
allophanate + H2O	Pseudomonas sp.	the reverse reaction is catalyzed by urea carboxylase	2 CO2 + 2 NH3	-	ir

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas sp.	Q936X2	gene atzF	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant active His-tagged enzyme from Escherichia coli strain BL21 (lambdaDE3) by nickel affinity chromatography and gel filtration to about 98% purity and apparent homogeneity	Pseudomonas sp.

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
allophanate + H2O	-	Pseudomonas sp.	2 CO2 + 2 NH3	-	ir
allophanate + H2O	the reverse reaction is catalyzed by urea carboxylase	Pseudomonas sp.	2 CO2 + 2 NH3	-	ir

Subunits

Subunits	Comment	Organism
homohexamer	6 * 50000, recombinant enzyme, SDS-PAGE	Pseudomonas sp.

Synonyms

Synonyms	Comment	Organism
AtzF	-	Pseudomonas sp.

General Information

General Information	Comment	Organism
evolution	allophanate hydrolase is a member of the amidase family of enzymes that possess a conserved serine- and glycine-rich motif, the so-called amidase signature sequence	Pseudomonas sp.
metabolism	allophanate hydrolase also participates in the cyanuric acid mineralization pathway, in which the cyanuric acid ring is hydrolytically opened by cyanuric acid hydrolase (AtzD or TrzD, EC 3.5.2.15) forming the unstable metabolite carboxybiuret, which spontaneously decarboxylates to form biuret. Allophanate is produced from biuret by AtzE (biuret hydrolase; EC 3.5.1.84) via a single deamination. Hydrolysis of allophanate is then carried out by allophanate hydrolase. Both pathways, cyanuric acid mineralization pathway and urea carboxylase pathway, depend upon allophanate deamination by allophanate hydrolase to avoid spontaneous decarboxylation (and urea formation)	Pseudomonas sp.