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Literature summary for 3.5.1.87 extracted from

  • Martinez-Rodriguez, S.; Clemente-Jimenez, J.M.; Rodriguez-Vico, F.; Las Heras-Vazquez, F.J.
    Molecular cloning and biochemical characterization of L-N-carbamoylase from Sinorhizobium meliloti CECT4114 (2005), J. Mol. Microbiol. Biotechnol., 9, 16-25.
    View publication on PubMed
Search for more literature for 3.5.1.87

Activating Compound

Activating Compound Comment Organism Structure
Co2+ 2 mM, specific activity 2.27, relative activity 339% Sinorhizobium meliloti
Fe2+ 2 mM, specific activity 2.03, relative activity 303% Sinorhizobium meliloti
Mn2+ 2 mM, specific activity 3.24, relative activity 484% Sinorhizobium meliloti
Ni2+ 2 mM, specific activity 9.79, relative activity 1461% Sinorhizobium meliloti

Application

Application Comment Organism
biotechnology production of optically pure L-amino acids by an enzymatic method named hydantoinase process Sinorhizobium meliloti

Cloned(Commentary)

Cloned (Comment) Organism
the L-N-carbamoylase gene is amplified by PCR and cloned into the pBSK vector for expression of the protein in Escherichia coli BL21(DE3) cells Sinorhizobium meliloti

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis-(2-nitrobenzoic acid) specific activity 0.39, relative activity 58% Sinorhizobium meliloti
beta-mercaptoethanol specific activity 0.44, relative activity 66% Sinorhizobium meliloti
Cu2+ 2 mM, specific activity 0.2, relative activity 30% Sinorhizobium meliloti
EDTA 10 mM, specific activity 0, relative activity 0% Sinorhizobium meliloti
Fe3+ 2 mM, specific activity 0.44, relative activity 66% Sinorhizobium meliloti
Hg2+ 2 mM, specific activity 0, relative activity 0% Sinorhizobium meliloti
K+ 2 mM, specific activity 0.56, relative activity 84% Sinorhizobium meliloti
Na+ 2 mM, specific activity 0.57, relative activity 85% Sinorhizobium meliloti
Pb2+ 2 mM, specific activity 0.39, relative activity 58% Sinorhizobium meliloti
Zn2+ 2 mM, specific activity 0.29, relative activity 43% Sinorhizobium meliloti

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.65
-
 N-carbamoyl-L-tryptophan determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
0.69
-
 N-carbamoyl-L-methionine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
0.94
-
 N-carbamoyl-L-alanine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
2.61
-
 N-carbamoyl-L-phenylalanine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
4.8
-
 N-carbamoyl-L-tyrosine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
4.91
-
 N-carbamoyl-L-cysteine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
5.5
-
 N-acetyl-L-methionine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
12.9
-
 N-formyl-L-methionine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
34.47
-
 N-carbamoyl-L-valine determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti
51.23
-
 N-carbamoyl-L-glutamic acid determined at 40°C for 20 min at pH 8.0 after preincubation with 2 mM Ni2+ Sinorhizobium meliloti

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42000
-
 SDS-PAGE Sinorhizobium meliloti
44860
-
 molecular mass deduced from the amino acid sequence Sinorhizobium meliloti
90000
-
 gel filtration Sinorhizobium meliloti

Organism

Organism UniProt Comment Textmining
Sinorhizobium meliloti
-
-
-

Purification (Commentary)

Purification (Comment) Organism
a one-step purification procedure of the recombinant L-N-carbamoylase fused to the HIS6-tag is employed using immobilized cobalt affinity chromatography Sinorhizobium meliloti

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.66
-
 N-carbamoyl-L-methionine Sinorhizobium meliloti

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetyl-L-methionine + H2O
-
 Sinorhizobium meliloti L-methionine + acetate
-
 ?
N-carbamoyl-L-alanine + H2O
-
 Sinorhizobium meliloti L-alanine + CO2 + NH3
-
 ?
N-carbamoyl-L-cysteine + H2O
-
 Sinorhizobium meliloti L-cysteine + CO2 + NH3
-
 ?
N-carbamoyl-L-glutamic acid + H2O
-
 Sinorhizobium meliloti L-glutamic acid + CO2 + NH3
-
 ?
N-carbamoyl-L-methionine + H2O activity assay Sinorhizobium meliloti L-methionine + CO2 + NH3
-
 ?
N-carbamoyl-L-phenylalanine + H2O
-
 Sinorhizobium meliloti L-phenylalanine + CO2 + NH3
-
 ?
N-carbamoyl-L-tryptophan + H2O
-
 Sinorhizobium meliloti L-tryptophan + CO2 + NH3
-
 ?
N-carbamoyl-L-tyrosine + H2O
-
 Sinorhizobium meliloti L-tyrosine + CO2 + NH3
-
 ?
N-carbamoyl-L-valine + H2O
-
 Sinorhizobium meliloti L-valine + CO2 + NH3
-
 ?
N-formyl-L-methionine + H2O
-
 Sinorhizobium meliloti L-methionine + HCOOH
-
 ?

Subunits

Subunits Comment Organism
homodimer
-
 Sinorhizobium meliloti

Synonyms

Synonyms Comment Organism
L-N-carbamoylase
-
 Sinorhizobium meliloti
N-carbamoyl-L-amino-acid hydrolase
-
 Sinorhizobium meliloti

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
 activity assay Sinorhizobium meliloti

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 70
-
 Sinorhizobium meliloti

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 activity assay Sinorhizobium meliloti

pH Range

pH Minimum pH Maximum Comment Organism
6 11
-
 Sinorhizobium meliloti