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Literature summary for 1.17.1.4 extracted from
- Purification and properties of xanthine dehydrogenase from Streptomyces cyanogenus (1979), J. Biochem., 86, 45-53.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| KCl | strong activation at 50 mM | Streptomyces cyanogenus |  |
| NaCl | nearly 2fold activation at 100 mM | Streptomyces cyanogenus | |
| NH4Cl | 2.5fold activation at 100 mM | Streptomyces cyanogenus | |
General Stability
| General Stability | Organism |
|---|---|
| enzyme from crude extract can be stabilized by addition of nicotinamide, cysteine and EDTA, purified enzyme more stable | Streptomyces cyanogenus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-chloromercuribenzoate | 89.2% inhibition of hypoxanthine oxidation at 1 mM | Streptomyces cyanogenus | |
| 5,5-dithiobis-(2-nitrobenzoate) | 45.4% inhibition of hypoxanthine oxidation at 1 mM | Streptomyces cyanogenus | |
| adenine | 62.8% inhibition of hypoxanthine oxidation at 0.5 mM | Streptomyces cyanogenus | |
| EDTA | 19.8% inhibition of hypoxanthine oxidation at 10 mM | Streptomyces cyanogenus | |
| iodoacetic acid | 21% inhibition at 1 mM | Streptomyces cyanogenus | |
| KCN | complete inhibition of hypoxanthine oxidation at 1 mM | Streptomyces cyanogenus | |
| o-phenanthroline | 19.5% inhibition of hypoxanthine oxidation at 10 mM | Streptomyces cyanogenus | |
| Quinacrine | 29.7% inhibition of hypoxanthine oxidation at 1 mM | Streptomyces cyanogenus | |
| Tiron | 36.8% inhibition of hypoxanthine oxidation at 10 mM | Streptomyces cyanogenus | |
| Urate | 36.6% inhibition of hypoxanthine oxidation at 0.5 mM | Streptomyces cyanogenus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.015 | - |
guanine | - |
Streptomyces cyanogenus | |
| 0.055 | - |
hypoxanthine | - |
Streptomyces cyanogenus | |
| 0.11 | - |
NAD+ | - |
Streptomyces cyanogenus | |
| 0.15 | - |
xanthine | - |
Streptomyces cyanogenus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Fe | - |
Streptomyces cyanogenus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 67000 | - |
2 * 67000, SDS-PAGE | Streptomyces cyanogenus |
| 125000 | - |
gel filtration, sedimentation equilibrium analysis | Streptomyces cyanogenus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2 2-hydroxypurine + 2 NAD+ + 2 H2O | Streptomyces cyanogenus | considerable activity | xanthine + 2,8-dihydroxypurine + 2 NADH + 2 H+ | 84% xanthine, 8% 2,8-dihydroxypurine formed | ? | |
| 2 hypoxanthine + 2 NAD+ + 2 H2O | Streptomyces cyanogenus | preferred substrate | xanthine + 6,8-dihydroxypurine + 2 NADH + 2 H+ | 100% xanthine, 51% 6,8-dihydroxypurine formed | ? | |
| 3 purine + 3 NAD+ + 3 H2O | Streptomyces cyanogenus | poor substrate | hypoxanthine + 8-hydroxypurine + 2-hydroxypurine + 3 NADH + 3 H+ | 2.3% hypoxanthine, 2.3% 8-hydroxypurine and traces of 2-hydroxypurine formed | ? | |
| 6,8-dihydroxypurine + NAD+ + H2O | Streptomyces cyanogenus | - |
urate + NADH | 18% urate formed | ? | |
| guanine + NAD+ + H2O | Streptomyces cyanogenus | 81.3% of the activity compared to hypoxanthine | ? + NADH | - |
? | |
| hypoxanthine + NADP+ + H2O | Streptomyces cyanogenus | 2.4% of the activity compared to NAD+ | xanthine + NADPH | - |
? | |
| hypoxanthine + O2 | Streptomyces cyanogenus | very low activity | xanthine + H2O2 | - |
? | |
| xanthine + NAD+ + H2O | Streptomyces cyanogenus | 61% of the activity compared to hypoxanthine | urate + NADH | 91% urate formed | ? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces cyanogenus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| to homogeneity, chromatography techniques | Streptomyces cyanogenus |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| cell culture | - |
Streptomyces cyanogenus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 10.6 | - |
- |
Streptomyces cyanogenus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2 2-hydroxypurine + 2 NAD+ + 2 H2O | considerable activity | Streptomyces cyanogenus | xanthine + 2,8-dihydroxypurine + 2 NADH + 2 H+ | 84% xanthine, 8% 2,8-dihydroxypurine formed | ? | |
| 2 hypoxanthine + 2 NAD+ + 2 H2O | preferred substrate | Streptomyces cyanogenus | xanthine + 6,8-dihydroxypurine + 2 NADH + 2 H+ | 100% xanthine, 51% 6,8-dihydroxypurine formed | ? | |
| 3 purine + 3 NAD+ + 3 H2O | poor substrate | Streptomyces cyanogenus | hypoxanthine + 8-hydroxypurine + 2-hydroxypurine + 3 NADH + 3 H+ | 2.3% hypoxanthine, 2.3% 8-hydroxypurine and traces of 2-hydroxypurine formed | ? | |
| 6,8-dihydroxypurine + NAD+ + H2O | - |
Streptomyces cyanogenus | urate + NADH | 18% urate formed | ? | |
| acetaldehyde + NAD+ + H2O | very low activity | Streptomyces cyanogenus | acetate + NADH + H+ | - |
? | |
| guanine + NAD+ + H2O | 81.3% of the activity compared to hypoxanthine | Streptomyces cyanogenus | ? + NADH | - |
? | |
| hypoxanthine + 2,6-dichlorophenolindophenol + H2O | 12.2% of the activity compared to NAD+ | Streptomyces cyanogenus | xanthine + ? | - |
? | |
| hypoxanthine + cytochrome c + H2O | 2.1% of the activity compared to NAD+ | Streptomyces cyanogenus | xanthine + reduced cytochrome c | - |
? | |
| hypoxanthine + ferricyanide + H2O | 17.2% of the activity compared to NAD+ | Streptomyces cyanogenus | xanthine + ferrocyanide | - |
? | |
| hypoxanthine + NADP+ + H2O | 2.4% of the activity compared to NAD+ | Streptomyces cyanogenus | xanthine + NADPH | - |
? | |
| hypoxanthine + nitroblue tetrazolium | 18% of the activity compared to NAD+ | Streptomyces cyanogenus | xanthine + ? | - |
? | |
| hypoxanthine + O2 | very low activity | Streptomyces cyanogenus | xanthine + H2O2 | - |
? | |
| inosine + NAD+ + H2O | low activity | Streptomyces cyanogenus | ? + NADH | - |
? | |
| propionaldehyde + NAD+ + H2O | low activity | Streptomyces cyanogenus | propionic acid + NADH | - |
? | |
| theobromine + NAD+ + H2O | very low activity | Streptomyces cyanogenus | ? + NADH | - |
? | |
| theophylline + NAD+ + H2O | low activity | Streptomyces cyanogenus | ? + NADH | - |
? | |
| xanthine + NAD+ + H2O | 61% of the activity compared to hypoxanthine | Streptomyces cyanogenus | urate + NADH | 91% urate formed | ? | |
| xanthosine + NAD+ + H2O | 15.1% of the activity compared to hypoxanthine | Streptomyces cyanogenus | ? + NADH | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 67000, SDS-PAGE | Streptomyces cyanogenus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
- |
Streptomyces cyanogenus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | - |
up to 150% activity when kept 24-120 h in a pH range of 9-11 | Streptomyces cyanogenus |
| 55 | - |
stable below | Streptomyces cyanogenus |
| 65 | - |
complete loss of activity in 10 min | Streptomyces cyanogenus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.7 | - |
- |
Streptomyces cyanogenus |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 12 | stable for 24 h if stored at 4°C | Streptomyces cyanogenus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | - |
Streptomyces cyanogenus | |
| additional information | iron-containing flavoprotein | Streptomyces cyanogenus | |
| additional information | molar ratio of FAD to iron to labile sulfide per mol enzyme is 2 : 14 : 2 | Streptomyces cyanogenus | |
| NAD+ | cannot be replaced by NADP+ | Streptomyces cyanogenus | |
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