Cloned (Comment) | Organism |
---|---|
gene ndmA, recombinant expression of ndmA and ndmD genes in Escherichia coli | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
additional information | production of theobromine and paraxanthine by engineered Escherichia coli expressing the enzyme NdmA and NdmN from Psseudomonas putida CBS5, method evaluation and optimization. Addition of metal ions is not required, additon of Co2+ and Zn2+ might inhibit the process | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | - |
Caffeine | pH and temperature not specified in the publication | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7-methylxanthine + O2 + NADH + H+ | Pseudomonas putida | - |
xanthine + NAD+ + H2O + formaldehyde | - |
? | |
7-methylxanthine + O2 + NADH + H+ | Pseudomonas putida CBB5 | - |
xanthine + NAD+ + H2O + formaldehyde | - |
? | |
caffeine + O2 + NADH + H+ | Pseudomonas putida | - |
theobromine + NAD+ + H2O + formaldehyde | - |
? | |
caffeine + O2 + NADH + H+ | Pseudomonas putida CBB5 | - |
theobromine + NAD+ + H2O + formaldehyde | - |
? | |
additional information | Pseudomonas putida | enzyme NdmA catalyzes NADH-dependent N1-demethylation of caffeine to theobromine and theophylline to 3-methylxanthine, and subsequently to 7-methylxanthine, and xanthine. Theobromine is the major product due to highly selective N1-demethylation caffeine by NdmA. But the minor production of paraxanthine from caffeine is due to slight promiscuity of the NdmA towards the N3-position on the caffeine molecule. Direct N-demethylation of caffeine at the N3 position does not occur appreciably with the three N-demethylases involved in caffeine degradation in CBB5 | ? | - |
? | |
additional information | Pseudomonas putida CBB5 | enzyme NdmA catalyzes NADH-dependent N1-demethylation of caffeine to theobromine and theophylline to 3-methylxanthine, and subsequently to 7-methylxanthine, and xanthine. Theobromine is the major product due to highly selective N1-demethylation caffeine by NdmA. But the minor production of paraxanthine from caffeine is due to slight promiscuity of the NdmA towards the N3-position on the caffeine molecule. Direct N-demethylation of caffeine at the N3 position does not occur appreciably with the three N-demethylases involved in caffeine degradation in CBB5 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | H9N289 | - |
- |
Pseudomonas putida CBB5 | H9N289 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7-methylxanthine + O2 + NADH + H+ | - |
Pseudomonas putida | xanthine + NAD+ + H2O + formaldehyde | - |
? | |
7-methylxanthine + O2 + NADH + H+ | - |
Pseudomonas putida CBB5 | xanthine + NAD+ + H2O + formaldehyde | - |
? | |
caffeine + O2 + NADH + H+ | - |
Pseudomonas putida | theobromine + NAD+ + H2O + formaldehyde | - |
? | |
caffeine + O2 + NADH + H+ | - |
Pseudomonas putida CBB5 | theobromine + NAD+ + H2O + formaldehyde | - |
? | |
additional information | enzyme NdmA catalyzes NADH-dependent N1-demethylation of caffeine to theobromine and theophylline to 3-methylxanthine, and subsequently to 7-methylxanthine, and xanthine. Theobromine is the major product due to highly selective N1-demethylation caffeine by NdmA. But the minor production of paraxanthine from caffeine is due to slight promiscuity of the NdmA towards the N3-position on the caffeine molecule. Direct N-demethylation of caffeine at the N3 position does not occur appreciably with the three N-demethylases involved in caffeine degradation in CBB5 | Pseudomonas putida | ? | - |
? | |
additional information | enzyme NdmA catalyzes NADH-dependent N1-demethylation of caffeine to theobromine and theophylline to 3-methylxanthine, and subsequently to 7-methylxanthine, and xanthine. Theobromine is the major product due to highly selective N1-demethylation caffeine by NdmA. But the minor production of paraxanthine from caffeine is due to slight promiscuity of the NdmA towards the N3-position on the caffeine molecule. Direct N-demethylation of caffeine at the N3 position does not occur appreciably with the three N-demethylases involved in caffeine degradation in CBB5 | Pseudomonas putida CBB5 | ? | - |
? | |
theophylline + O2 + NADH + H+ | - |
Pseudomonas putida | 3-methylxanthine + NAD+ + H2O + formaldehyde | - |
? | |
theophylline + O2 + NADH + H+ | - |
Pseudomonas putida CBB5 | 3-methylxanthine + NAD+ + H2O + formaldehyde | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ndmA | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
in vivo assay, recombinant enzyme | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.17 | - |
Caffeine | pH and temperature not specified in the publication | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase that requires a partner reductase, NdmD, to transfer electrons from NADH | Pseudomonas putida | |
[2Fe-2S]-center | NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase that requires a partner reductase, NdmD, to transfer electrons from NADH | Pseudomonas putida |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme NdmA catalyzes NADH-dependent N1-demethylation of caffeine to theobromine and theophylline to 3-methylxanthine, and subsequently to 7-methylxanthine, and xanthine. The oxidoreductase NdmD, UniProt ID H9N291, catalyzes the oxidation of NADH and transfers electrons to NdmA and NdmB, which catalyze the N-demethylation reaction | Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
85.68 | - |
Caffeine | pH and temperature not specified in the publication | Pseudomonas putida |