Literature summary for 3.5.1.54 extracted from

Zhao, J.; Zhu, L.; Fan, C.; Wu, Y.; Xiang, S.
Structure and function of urea amidolyase (2018), Biosci. Rep., 38, BSR20171617 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene KLLA0_E08119g, phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)Star	Kluyveromyces lactis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme, sitting drop vapor diffusion method, mixing of protein in 0.5 mM urea, 0.5 mM ADP, and 0.5 mM sodium malonate, pH 7.0, with reservoir solution containing 0.1 M Tris/HCl, pH 7.5, 0.2 M ammonium sulfate, 12% PEG 8000, and 2% PEG 3350, X-ray diffraction structure determination and analysis at 6.5 A resolution, modelling by molecular replacement method using the structures of KlUC (PDB 3VA7) and KlAH (PDB 4ISS) as search models	Kluyveromyces lactis

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme, sitting drop vapor diffusion method, mixing of protein in 0.5 mM urea, 0.5 mM ADP, and 0.5 mM sodium malonate, pH 7.0, with reservoir solution containing 0.1 M Tris/HCl, pH 7.5, 0.2 M ammonium sulfate, 12% PEG 8000, and 2% PEG 3350, X-ray diffraction structure determination and analysis at 6.5 A resolution, modelling by molecular replacement method using the structures of KlUC (PDB 3VA7) and KlAH (PDB 4ISS) as search models

Kluyveromyces lactis

Protein Variants

Protein Variants	Comment	Organism
G559E/G572E	site-directed mutagenesis, the mutant shows abolished activity	Kluyveromyces lactis
additional information	generation of the KlUADELTABCCP construct via insertion a stop codon after base pair 5223	Kluyveromyces lactis
S177A	site-directed mutagenesis, the mutation inactivates the isolated KlAH	Kluyveromyces lactis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
400000	-	gel filtration	Kluyveromyces lactis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
urea-1-carboxylate + H2O	Kluyveromyces lactis	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis CBS 2359	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis NRRL Y-1140	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis DSM 70799	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis WM37	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis ATCC 8585	-	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	Kluyveromyces lactis NBRC 1267	-	2 CO2 + 2 NH3	-	?

Organism

Organism	UniProt	Comment	Textmining
Kluyveromyces lactis	Q6CP22	-	-
Kluyveromyces lactis ATCC 8585	Q6CP22	-	-
Kluyveromyces lactis CBS 2359	Q6CP22	-	-
Kluyveromyces lactis DSM 70799	Q6CP22	-	-
Kluyveromyces lactis NBRC 1267	Q6CP22	-	-
Kluyveromyces lactis NRRL Y-1140	Q6CP22	-	-
Kluyveromyces lactis WM37	Q6CP22	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)Star by nickel affinity chromatography and gel filtration	Kluyveromyces lactis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis CBS 2359	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis NRRL Y-1140	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis DSM 70799	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis WM37	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis ATCC 8585	?	-	?
additional information	allophanate is produced at the active site of the UC C-terminal domain and is translocated to that of the AH N-domain for subsequent reaction. Allophanate is translocated from the active site of UC C-terminal domain to that of the AH N-domain via diffusion through solvent, instead of being channeled through the dimer	Kluyveromyces lactis NBRC 1267	?	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis CBS 2359	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis NRRL Y-1140	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis DSM 70799	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis WM37	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis ATCC 8585	2 CO2 + 2 NH3	-	?
urea-1-carboxylate + H2O	-	Kluyveromyces lactis NBRC 1267	2 CO2 + 2 NH3	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 190000, recombinant enzyme, SDS-PAGE	Kluyveromyces lactis
More	analysis of interactions between the KlUA monomers	Kluyveromyces lactis
More	urea amidolyase is composed of urea carboxylase (UC) and allophanate hydrolase (AH) domains. KlUC and KlAH are monomeric and dimeric in solution, respectively. The relatively smaller UC-AH interface therefore does not play a major role in the UA holo-enzyme assembly. In the isolated KlAH, the active sites are located near the dimer interface. The extensive interactions at the dimer interface most likely stabilize the structure of the active sites. Consistent with this, the G559E/G572E mutation that renders the isolated KlAH monomeric severely inhibited its activity	Kluyveromyces lactis

Synonyms

Synonyms	Comment	Organism
KlAH	-	Kluyveromyces lactis
KLLA0_E08119g	-	Kluyveromyces lactis
urea amidolyase	-	Kluyveromyces lactis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Kluyveromyces lactis

General Information

General Information	Comment	Organism
metabolism	urea amidolyase catalyzes the conversion of urea to ammonium, the essential first step in utilizing urea as a nitrogen source	Kluyveromyces lactis
additional information	urea amidolyase is composed of urea carboxylase (UC) and allophanate hydrolase (AH) domains. UC converts urea to allophanate, and AH subsequently converts it to ammonium. The AH domain is composed of N- and C-domains, which catalyze sequential reactions in the allophanate to ammonium conversion	Kluyveromyces lactis