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Literature summary extracted from

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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.54 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)

EC Number Crystallization (Comment) Organism
3.5.1.54 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

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.54 400000
-
 gel filtration Kluyveromyces lactis

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.54 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 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 ?
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis CBS 2359 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis NRRL Y-1140 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis DSM 70799 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis WM37 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis ATCC 8585 2 CO2 + 2 NH3
-
 ?
3.5.1.54 urea-1-carboxylate + H2O
-
 Kluyveromyces lactis NBRC 1267 2 CO2 + 2 NH3
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.54 homodimer 2 * 190000, recombinant enzyme, SDS-PAGE Kluyveromyces lactis
3.5.1.54 More analysis of interactions between the KlUA monomers Kluyveromyces lactis
3.5.1.54 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

EC Number Synonyms Comment Organism
3.5.1.54 KlAH
-
 Kluyveromyces lactis
3.5.1.54 KLLA0_E08119g
-
 Kluyveromyces lactis
3.5.1.54 urea amidolyase
-
 Kluyveromyces lactis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.54 8
-
 assay at Kluyveromyces lactis

General Information

EC Number General Information Comment Organism
3.5.1.54 metabolism urea amidolyase catalyzes the conversion of urea to ammonium, the essential first step in utilizing urea as a nitrogen source Kluyveromyces lactis
3.5.1.54 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