EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.3.17 | expression of wild-type and mutants in Escherichia coli strain Rosetta(DE3)pLysS | Arthrobacter globiformis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.1.3.17 | purified recombinnat wild-type enzyme and mutant V464A, hanging drop vapor diffusion, room temperature, from 10-15% v/v PEG 6000, 50-200 mM magnesium acetate, 200 mM trimethylamine, and 0.08 M sodium cacodylate, pH 6.0, X-ray diffraction structure determination and analysis at 2.2 A resolution | Arthrobacter globiformis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.3.17 | V464A | site-directed mutagenesis, mutation of the residue near the flavin C(4a) atom and the hydrophobic cavity, replacement of Val464 with alanine or threonine does not affect the reductive half-reaction, but it reduces the oxidative half-reaction by about 50fold, and the 3D structure of the Val464Ala enzyme is essentially identical to that of the wild-type enzyme | Arthrobacter globiformis |
1.1.3.17 | V464T | site-directed mutagenesis, mutation of the residue near the flavin C(4a) atom and the hydrophobic cavity, replacement of Val464 with alanine or threonine does not affect the reductive half-reaction, but it reduces the oxidative half-reaction by about 50fold | Arthrobacter globiformis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.3.17 | additional information | - |
additional information | minimal kinetic mechanism for reductive half-reaction of the V464A and V464T mutant enzymes, oxidative half-reactions' steady-state kinetic mechanisms, overview | Arthrobacter globiformis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.3.17 | choline + 2 O2 + H2O2 | Arthrobacter globiformis | - |
betaine + 2 H2O2 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.3.17 | Arthrobacter globiformis | Q7X2H8 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.3.17 | recombinant wild-type and mutants from Escherichia coli strain Rosetta(DE3)pLysS | Arthrobacter globiformis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.3.17 | 3,3-dimethylbutan-1-ol + 2 O2 + H2O2 | - |
Arthrobacter globiformis | ? | - |
? | |
1.1.3.17 | choline + 2 O2 + H2O2 | - |
Arthrobacter globiformis | betaine + 2 H2O2 | - |
? | |
1.1.3.17 | choline + 2 O2 + H2O2 | two-step oxidation of choline with formation of betaine aldehyde as intermediate, the overall reaction consists of oxidative and reductive half-reactions | Arthrobacter globiformis | betaine + 2 H2O2 | - |
? |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.3.17 | 25 | - |
assay at | Arthrobacter globiformis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.3.17 | 10 | - |
assay at | Arthrobacter globiformis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.3.17 | FAD | required for catalysis, covalently bound to the Nepsilon2 atom of His99 of the wild-type enzyme | Arthrobacter globiformis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.3.17 | additional information | the side chain of Val464 in the active site cavity in choline oxidase provides a nonpolar site that is required to guide oxygen in proximity of the C(4a) atom of the flavin, where it will subsequently react via electrostatic catalysis | Arthrobacter globiformis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.3.17 | additional information | - |
additional information | kcat/Koxygen values are independent of the pH between pH 5.0 and 10.0, average value is 1.7 mM | Arthrobacter globiformis | |
1.1.3.17 | 0.031 | - |
3,3-dimethylbutan-1-ol | pH 8.0, 25°C, wild-type enzyme | Arthrobacter globiformis | |
1.1.3.17 | 0.031 | - |
3,3-dimethylbutan-1-ol | pH 8.0, 25°C, mutant V464A | Arthrobacter globiformis | |
1.1.3.17 | 0.1 | - |
O2 | kcat/Kox, pH 10.0, 25°C, mutant V464A | Arthrobacter globiformis | |
1.1.3.17 | 0.125 | - |
O2 | kcat/Kox, pH 10.0, 25°C, mutant V464T | Arthrobacter globiformis | |
1.1.3.17 | 1.9 | - |
O2 | kcat/Kox, pH 10.0, 25°C, wild-type enzyme | Arthrobacter globiformis | |
1.1.3.17 | 2 | - |
3,3-dimethylbutan-1-ol | pH 8.0, 25°C, wild-type enzyme | Arthrobacter globiformis |