Information on EC 2.1.1.159 - theobromine synthase

Word Map on EC 2.1.1.159
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.1.1.159
-
RECOMMENDED NAME
GeneOntology No.
theobromine synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + 7-methylxanthine = S-adenosyl-L-homocysteine + 3,7-dimethylxanthine
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
caffeine biosynthesis I
-
-
theobromine biosynthesis I
-
-
Caffeine metabolism
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:7-methylxanthine N3-methyltransferase
This is the third enzyme in the caffeine-biosynthesis pathway. This enzyme can also catalyse the conversion of paraxanthine into caffeine, although the paraxanthine pathway is considered to be a minor pathway for caffeine biosynthesis [2,3].
CAS REGISTRY NUMBER
COMMENTARY hide
72270-63-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Coffea sp.
-
-
-
Manually annotated by BRENDA team
variant Paullinia cupana sorbilis, cultivars BRS-Amazonas, BRS-Maues, BRS-Luzeia, BRS-CG372, and BRS-CG611
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
Coffea sp.
-
the enzyme catalyzes the second step in caffeine biosynthesis, pathway overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + 1,7-dimethylxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
-
i.e. paraxanthine
i.e. caffeine
-
?
S-adenosyl-L-methionine + 1-methylxanthine
S-adenosyl-L-homocysteine + 1,3-dimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + 3,7-dimethylxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + 3-methylxanthine
S-adenosyl-L-homocysteine + 1,3-dimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + 7-methylxanthine
S-adenosyl-L-homocysteine + 3,7-dimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + 7-methylxanthine
S-adenosyl-L-homocysteine + 3,7-dimethylxanthine + H+
show the reaction diagram
S-adenosyl-L-methionine + paraxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + 1,7-dimethylxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
-
i.e. paraxanthine
i.e. caffeine
-
?
S-adenosyl-L-methionine + 3,7-dimethylxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + 7-methylxanthine
S-adenosyl-L-homocysteine + 3,7-dimethylxanthine
show the reaction diagram
S-adenosyl-L-methionine + paraxanthine
S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
Coffea sp.
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chloromercuribenzoate
-
complete inhibition at 0.5 mM, 30% inhibition at 0.05 mM
azouracil
-
20% inhibition at 0.1 mM, 28% inhibition at 1 mM
iodoacetate
-
complete inhibition at 100 mM, 79% inhibition at 10 mM
KCN
-
88% inhibition at 100 mM, 44% inhibition at 10 mM
N-Methylmaleimide
-
complete inhibition at 1 mM, 17% inhibition at 0.1 mM
NaN3
-
39% inhibition at 10 mM, 10% at 1 mM
additional information
-
no or poor inhibition by EDTA at 0.5-5 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
paraxanthine
-
i.e. 1,7-dimethylxanthine, slight stimulation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.258 - 0.277
3,7-Dimethylxanthine
0.0072 - 0.328
7-methylxanthine
0.031 - 0.738
paraxanthine
0.01 - 0.014
S-adenosyl-L-methionine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.09
recombinant enzyme in Escherichia coli cell extract, substrate 7-methylxanthine
4.59
recombinant CCS1
15.45
-
purified enzyme
95
-
purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.9
-
assay at
8
assay at; assay at
8.3
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 5.1
-
chromatofocusing
5.1
-
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
isozyme MXMT2; isozyme MXTM1
Manually annotated by BRENDA team
; low expression level of MXMT
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
co-localization of xanthosine methyltransferase, 7-methylxanthine methyltransferase, and 3,7-dimethylxanthine methyltransferase
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
61000
-
gel filtration
67000
-
about, gel filtration
74000
recombinant isozyme MXMT2, gel filtration
80000
about, recombinant enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer or dimer
-
x * 41000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant wild-type and selenomethionine-labeled DXMT, 23-28% PEG 3350, 0.2 M LiCl, 0.1 M Tris-HCl, pH 8.5-8.7, 2 mM DTT, 1 mM S-adenosyl-L-cysteine, and 1 mM theobromine, 1-3 days, 20C, X-ray diffraction structure determination and analysis at 2.5-2.7 A resolution
purified recombinant wild-type and selenomethionine-labeled DXMT, 23-28% PEG 3350, 0.2 M LiCl, 0.1 M Tris-HCl, pH 8.5-8.7, 2 mM DTT, 2 mM S-adenosyl-L-cysteine, and 2 mM theobromine, 1-3 days, 20C, plate-like crystals, X-ray diffraction structure determination and analysis at 2.0-2.7 A resolution, molecular replacement
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 39fold from young leaves by ammonium sulfate fractionation, 3 steps of ion exchange chromatography, and gel filtration
-
native enzyme 523fold from young, developing leaves by ammonium sulfate fractionation, hydroxylapatite, anion exchange, and adenosine affinity chromatography, and gel filtration to homogeneity
-
partially purified by Ni-NTA agarose column chromatography
-
recombinant GST-fusion enzyme from Escherichia coli strain JM109 by glutathione affinity chromatography
recombinant His-tagged DXMT1 from Escherichia coli strain BL21(DE3) to homogeneity by nickel affinity chromatography, cleavage of the His-tag with tobacco etch virus, TEV, protease, followed by gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of MXMT vectors for Agrobacterium tumefaciens-mediated transformation
-
DNA and amino acid sequence determination and analysis, cloning of MXMT vectors for Agrobacterium tumefaciens-mediated transformation; gene MXMT2, DNA and amino acid sequence determination and analysis
DNA and amino acid sequence determination and analysis, expression in Escherichia coli as wild-type and as Trx-fusion protein
expressed in Escherichia coli BL21-pRil cells
-
expressed in Escherichia coli Rosetta BL21 (DE3) and Saccharomyces cerevisiae INVSC1 cells
expression in Escherichia coli
-
expression of His-tagged DXMT1 in Escherichia coli strain BL21(DE3)
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), sequence comparisonof caffeine synthetic enzymes from coffee
Coffea sp.
-
gene CaMTL2, cDNA library construction, DNA and amino acid sequence determination and analysis; gene CaMXMT, cDNA library construction, DNA and amino acid sequence determination and analysis, sequence comparison with other species, expression of CaMXMT as GST-fusion enzyme in Escherichia coli strain JM109
gene TCS1, cDNA and amino acid sequence determination and analysis, expression in Escherichia coli
isozyme MXMT1, DNA and amnio acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged isozymes in Escherichia coli Bl21(DE3); isozyme MXMT2, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged isozymes in Escherichia coli Bl21(DE3)
isozyme MXMT1, functional expression in transgenic Nicotiana tabacum plant leaves
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
naturally occuring low-caffeine fruits have a lower expression of the theobromine synthase and caffeine synthase genes and also contain an extra transcript of the caffeine synthase gene encoding a mutant form. The absence of caffeine in these mutants probably results from a combination of transcriptional regulation and the presence of mutations
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
expression of the caffeine biosynthesis enzymes in transgenic crop plants may protect against the crop damaging larvae of pests
biotechnology
large-scale production of transgenic enzyme-deficient Coffea arabica and Camellia sinensis plants are a practical possibilty for production of decaffeinated coffee or tea
food industry
large-scale production of transgenic enzyme-deficient Coffea arabica and Camellia sinensis plants are a practical possibilty for production of decaffeinated coffee or tea