Cloned (Comment) | Organism |
---|---|
gene tmuM, DNA and amino acid sequence determination and analysis, genes involved in the caffeine C-8 oxidation pathway are located in a 25.2-kb genomic DNA fragment of CBB1, including cdhABC (coding for caffeine dehydrogenase) and tmuM (coding for TmuM) | Pseudomonas sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0012 | - |
1-Methylurate | pH 7.5, 30°C | Pseudomonas sp. | |
0.0013 | - |
3,7-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
0.0102 | - |
1,3,7-trimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
0.1265 | - |
1,3-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,3,7-trimethylurate + NADH + H+ + O2 | Pseudomonas sp. | - |
1,3,7-trimethyl-5-hydroxyisourate + NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. | I7ASS7 | - |
- |
Purification (Comment) | Organism |
---|---|
native enzyme 55fold from strain CBB1 by anion exchange, affinity, and hydrophobic interaction chromatography, followed by gel filtration | Pseudomonas sp. |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.128 | - |
purified native enzyme, pH 7.5, 30°C | Pseudomonas sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,3,7-trimethylurate + NADH + H+ + O2 | - |
Pseudomonas sp. | 1,3,7-trimethyl-5-hydroxyisourate + NAD+ + H2O | - |
? | |
1,3-dimethylurate + NADH + H+ + O2 | - |
Pseudomonas sp. | 1,3-dimethyl-5-hydroxyisourate + NAD+ + H2O | - |
? | |
1-methylurate + NADH + H+ + O2 | low activity | Pseudomonas sp. | 1-methyl-5-hydroxyisourate + NAD+ + H2O | - |
? | |
3,7-dimethylurate + NADH + H+ + O2 | best substrate | Pseudomonas sp. | 3,7-dimethyl-5-hydroxyisourate + NAD+ + H2O | - |
? | |
additional information | enzyme TmuM prefers trimethyluric acids and, to a lesser extent, dimethyluric acids as substrates, but it exhibits no activity on uric acid | Pseudomonas sp. | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NADH-dependent trimethyluric acid monooxygenase | - |
Pseudomonas sp. |
tmuM | - |
Pseudomonas sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Pseudomonas sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.485 | - |
1-Methylurate | pH 7.5, 30°C | Pseudomonas sp. | |
1.97 | - |
3,7-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
3.08 | - |
1,3-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
7.48 | - |
1,3,7-trimethylurate | pH 7.5, 30°C | Pseudomonas sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | flavoprotein | Pseudomonas sp. | |
NADH | dependent on | Pseudomonas sp. |
General Information | Comment | Organism |
---|---|---|
evolution | genes involved in the caffeine C-8 oxidation pathway are located in a 25.2-kb genomic DNA fragment of CBB1, including cdhABC (coding for caffeine dehydrogenase) and tmuM (coding for TmuM), Physical map of genes for caffeine transformation in a 25.2-kb gene cluster in Pseudomonas sp. strain CBB1, overview | Pseudomonas sp. |
metabolism | the enzyme takes part in the caffeine C-8 oxidation pathway as the second enzyme, a NADH-dependent trimethyluric acid monooxygenase that catalyzes the conversion of trimethylurate to 1,3,7-trimethyl-5-hydroxyisourate. This product spontaneously decomposes to racemic 3,6,8-trimethylallantoin | Pseudomonas sp. |
additional information | homology models of trimethyluric acid monooxygenase against uric acid oxidase HpxO (which catalyzes uric acid to 5-hydroxyisourate) reveal a much bigger and hydrophobic cavity to accommodate the larger substrates | Pseudomonas sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
25 | - |
1,3-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
408.3 | - |
1-Methylurate | pH 7.5, 30°C | Pseudomonas sp. | |
735 | - |
1,3,7-trimethylurate | pH 7.5, 30°C | Pseudomonas sp. | |
1490 | - |
3,7-dimethylurate | pH 7.5, 30°C | Pseudomonas sp. |