BRENDA - Enzyme Database
show all sequences of 1.14.13.128

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)
Commentary
Organism
gene ndmA, recombinant expression of ndmA and ndmD genes in Escherichia coli
Pseudomonas putida
Engineering
Amino acid exchange
Commentary
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 [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.037
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase
Pseudomonas putida
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas putida
H9N289
-
-
Pseudomonas putida CBB5
H9N289
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
caffeine + O2 + NADH + H+
-
744193
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
744193
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
744193
Pseudomonas putida CBB5
?
-
-
-
-
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
in vivo assay, recombinant enzyme
Pseudomonas putida
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.17
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Cofactor
Cofactor
Commentary
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
Cloned(Commentary) (protein specific)
Commentary
Organism
gene ndmA, recombinant expression of ndmA and ndmD genes in Escherichia coli
Pseudomonas putida
Cofactor (protein specific)
Cofactor
Commentary
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
Engineering (protein specific)
Amino acid exchange
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.037
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
NdmA enzyme is a Rieske (2Fe-2S) non-heme iron monooxygenase
Pseudomonas putida
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
?
-
-
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
7-methylxanthine + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
xanthine + NAD+ + H2O + formaldehyde
-
-
-
?
caffeine + O2 + NADH + H+
-
744193
Pseudomonas putida
theobromine + NAD+ + H2O + formaldehyde
-
-
-
?
caffeine + O2 + NADH + H+
-
744193
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
744193
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
744193
Pseudomonas putida CBB5
?
-
-
-
-
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
theophylline + O2 + NADH + H+
-
744193
Pseudomonas putida CBB5
3-methylxanthine + NAD+ + H2O + formaldehyde
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
in vivo assay, recombinant enzyme
Pseudomonas putida
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.17
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
General Information
General Information
Commentary
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
General Information (protein specific)
General Information
Commentary
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 [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
85.68
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
85.68
-
Caffeine
pH and temperature not specified in the publication
Pseudomonas putida
Other publictions for EC 1.14.13.128
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
744193
Algharrawi
-
Production of theobromine by ...
Pseudomonas putida, Pseudomonas putida CBB5
Biocatal. Agricult. Biotechnol.
11
153-160
2017
-
-
1
-
1
-
-
1
-
1
-
6
-
4
-
-
-
-
-
-
-
-
8
-
1
-
-
1
-
-
-
2
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1
2
-
1
-
-
-
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1
-
1
-
6
-
-
-
-
-
-
-
-
8
-
1
-
-
1
-
-
-
-
-
1
1
-
1
1
727773
Summers
Caffeine junkie: an unpreceden ...
Pseudomonas putida, Pseudomonas putida CBB5
J. Bacteriol.
195
3933-3939
2013
-
-
1
-
-
-
-
1
-
1
-
4
-
3
-
-
1
-
-
-
-
-
4
-
-
-
-
1
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
1
-
1
-
4
-
-
-
1
-
-
-
-
4
-
-
-
-
1
-
-
-
-
-
-
-
-
1
1
722538
Summers
Novel, highly specific N-demet ...
Pseudomonas putida, Pseudomonas putida CBB5
J. Bacteriol.
194
2041-2049
2012
-
-
1
-
-
-
-
-
-
1
1
4
-
10
-
-
1
-
-
-
-
-
4
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
1
1
4
-
-
-
1
-
-
-
-
4
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
709854
Summers
Characterization of a broad-sp ...
Pseudomonas putida, Pseudomonas putida CBB5
Microbiology
157
583-592
2011
-
-
-
-
-
-
-
2
-
1
1
2
-
8
-
-
1
-
-
-
1
2
23
1
1
-
-
2
1
1
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
2
-
1
1
2
-
-
-
1
-
-
1
2
23
1
1
-
-
2
1
1
-
-
-
-
-
-
2
2
717407
Dash
Catabolic pathways and biotech ...
Pseudomonas putida
Biotechnol. Lett.
28
1993-2002
2006
-
2
-
-
-
-
1
1
-
-
1
4
-
7
-
-
1
-
-
-
-
1
7
1
2
-
-
-
2
-
-
1
-
-
-
-
2
-
1
-
-
-
-
1
-
1
-
-
1
4
-
-
-
1
-
-
-
1
7
1
2
-
-
-
2
-
-
-
-
2
2
-
-
-
707235
Glück
-
Heteroxanthinedemethylase, a n ...
Pseudomonas putida
Appl. Microbiol. Biotechnol.
28
59-62
1988
-
-
-
-
-
1
7
1
-
2
-
-
-
1
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1
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1
2
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1
-
1
-
1
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-
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1
-
7
-
1
-
2
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-
1
-
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1
2
-
1
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1
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1
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