Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Kluyveromyces lactis |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapor diffusion method, using 2.4 M ammonium sulfate and 0.1 M bis-Tris, pH 7.0 | Kluyveromyces lactis |
Protein Variants | Comment | Organism |
---|---|---|
D1321A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
D1584N | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
E1792A | the mutant shows reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
K1605A | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
Y1324F | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
Y1628F | the mutant shows severely reduced catalytic efficiency compared to the wild type enzyme | Kluyveromyces lactis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.32 | - |
Urea | wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
0.39 | - |
Urea | mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
1 | 1.3 | Urea | mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
7.7 | - |
Urea | mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
15 | - |
Urea | mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
78 | - |
Urea | mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
493 | - |
Urea | mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
137000 | - |
1 * 137000, calculated from amino acid sequence | Kluyveromyces lactis |
141000 | - |
gel filtration | Kluyveromyces lactis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + urea + HCO3- + H+ | Kluyveromyces lactis | - |
ADP + phosphate + urea-1-carboxylate | - |
? | |
ATP + urea + HCO3- + H+ | Kluyveromyces lactis ATCC 8585 | - |
ADP + phosphate + urea-1-carboxylate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Kluyveromyces lactis | Q6CP22 | - |
- |
Kluyveromyces lactis ATCC 8585 | Q6CP22 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography and Sephacryl S-300 gel filtration | Kluyveromyces lactis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + urea + HCO3- + H+ | - |
Kluyveromyces lactis | ADP + phosphate + urea-1-carboxylate | - |
? | |
ATP + urea + HCO3- + H+ | - |
Kluyveromyces lactis ATCC 8585 | ADP + phosphate + urea-1-carboxylate | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 137000, calculated from amino acid sequence | Kluyveromyces lactis |
monomer | 1 * 141000, dynamic light scattering | Kluyveromyces lactis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.44 | - |
Urea | mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
3.16 | - |
Urea | mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
45.8 | - |
Urea | mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
61.8 | - |
Urea | mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
64.9 | - |
Urea | mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
68.7 | - |
Urea | mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
210.8 | - |
Urea | wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Kluyveromyces lactis | |
biotin | - |
Kluyveromyces lactis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0029 | - |
Urea | mutant enzyme K1605A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
0.041 | - |
Urea | mutant enzyme D1321A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
4.3 | - |
Urea | mutant enzyme Y1628F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
5.5 | - |
Urea | mutant enzyme D1564N, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
5.9 | - |
Urea | mutant enzyme Y1324F, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
176 | - |
Urea | mutant enzyme E1792A, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis | |
659 | - |
Urea | wild type enzyme, in 200 mM Tris-HCl, pH 8.0, 0.001 mM EGTA, 3 mM magnesium chloride, 19.5 mM potassium chloride, at 22°C | Kluyveromyces lactis |