BRENDA - Enzyme Database

Production of theobromine by N-demethylation of caffeine using metabolically engineered E. coli

Algharrawi, K.; Summers, R.; Subramanian, M.; Biocatal. Agricult. Biotechnol. 11, 153-160 (2017)
No PubMed abstract available

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.13.128
gene ndmA, recombinant expression of ndmA and ndmD genes in Escherichia coli
Pseudomonas putida
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.14.13.128
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]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.128
0.037
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.13.128
Fe2+
NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase
Pseudomonas putida
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.13.128
7-methylxanthine + O2 + NADH + H+
Pseudomonas putida
-
xanthine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
7-methylxanthine + O2 + NADH + H+
Pseudomonas putida CBB5
-
xanthine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
Pseudomonas putida
-
theobromine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
Pseudomonas putida CBB5
-
theobromine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
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
?
-
-
-
1.14.13.128
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.13.128
Pseudomonas putida
H9N289
-
-
1.14.13.128
Pseudomonas putida CBB5
H9N289
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.128
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
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
744193
Pseudomonas putida
?
-
-
-
-
1.14.13.128
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
744193
Pseudomonas putida CBB5
?
-
-
-
-
1.14.13.128
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.14.13.128
30
-
in vivo assay, recombinant enzyme
Pseudomonas putida
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.13.128
3.17
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.128
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
1.14.13.128
[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
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.13.128
gene ndmA, recombinant expression of ndmA and ndmD genes in Escherichia coli
Pseudomonas putida
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.128
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
1.14.13.128
[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
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.14.13.128
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.128
0.037
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.13.128
Fe2+
NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase
Pseudomonas putida
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.14.13.128
7-methylxanthine + O2 + NADH + H+
Pseudomonas putida
-
xanthine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
7-methylxanthine + O2 + NADH + H+
Pseudomonas putida CBB5
-
xanthine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
Pseudomonas putida
-
theobromine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
Pseudomonas putida CBB5
-
theobromine + NAD+ + H2O + formaldehyde
-
-
?
1.14.13.128
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
?
-
-
-
1.14.13.128
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
?
-
-
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.128
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
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
744193
Pseudomonas putida
?
-
-
-
-
1.14.13.128
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
744193
Pseudomonas putida CBB5
?
-
-
-
-
1.14.13.128
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
1.14.13.128
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.14.13.128
30
-
in vivo assay, recombinant enzyme
Pseudomonas putida
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.14.13.128
3.17
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
General Information
EC Number
General Information
Commentary
Organism
1.14.13.128
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
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.14.13.128
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 [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.13.128
85.68
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.13.128
85.68
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida