BRENDA - Enzyme Database
show all sequences of 2.1.1.158

Essential region for 3-N methylation in N-methyltransferases involved in caffeine biosynthesis

Mizuno, K.; Kurosawa, S.; Yoshizawa, Y.; Kato, M.; Z. Naturforsch. C 65c, 257-265 (2010)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), sequence comparison of caffeine synthetic enzymes from coffee
Coffea sp.
Engineering
Protein Variants
Commentary
Organism
A23S
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/L191P/H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/Q161H/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows 60% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/Q161H/L191P/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/Q161H/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/Q161H/L191P/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
L191P
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
additional information
construction of several enzyme mutants, overview. The mutants of CmXRS1, that have 3-N methylation activity and produce caffeine from paraxanthine as a substrate, need to have replacement of the glutamine residue by histidine at position 161 in the CmXRS1 sequence, i.e. a Q161H mutation, overview
Coffea sp.
P104Q
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/L191P
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/Q161H
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/Q161H/L191P
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Q161H
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Q161H/L191P
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
required
Coffea sp.
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
S-adenosyl-L-methionine + xanthosine
Coffea sp.
-
S-adenosyl-L-homocysteine + 7-methylxanthosine
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Coffea sp.
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
additional information
7-methylxanthosine synthase shows the most critical substrate specifi city of the three types of N-methyltransferases in Coffea species
721124
Coffea sp.
?
-
-
-
-
S-adenosyl-L-methionine + xanthosine
-
721124
Coffea sp.
S-adenosyl-L-homocysteine + 7-methylxanthosine
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
CmXRS1
-
Coffea sp.
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Coffea sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Coffea sp.
Cofactor
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Coffea sp.
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), sequence comparison of caffeine synthetic enzymes from coffee
Coffea sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Coffea sp.
Engineering (protein specific)
Protein Variants
Commentary
Organism
A23S
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/L191P/H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/Q161H/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows 60% of wild-type 7-methylation activity
Coffea sp.
A23S/P104Q/Q161H/L191P/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/Q161H/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
A23S/Q161H/L191P/H219R
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
H219R
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
L191P
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
additional information
construction of several enzyme mutants, overview. The mutants of CmXRS1, that have 3-N methylation activity and produce caffeine from paraxanthine as a substrate, need to have replacement of the glutamine residue by histidine at position 161 in the CmXRS1 sequence, i.e. a Q161H mutation, overview
Coffea sp.
P104Q
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/L191P
site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/Q161H
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
P104Q/Q161H/L191P
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Q161H
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Q161H/L191P
site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity
Coffea sp.
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
required
Coffea sp.
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
S-adenosyl-L-methionine + xanthosine
Coffea sp.
-
S-adenosyl-L-homocysteine + 7-methylxanthosine
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
additional information
7-methylxanthosine synthase shows the most critical substrate specifi city of the three types of N-methyltransferases in Coffea species
721124
Coffea sp.
?
-
-
-
-
S-adenosyl-L-methionine + xanthosine
-
721124
Coffea sp.
S-adenosyl-L-homocysteine + 7-methylxanthosine
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
27
-
assay at
Coffea sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Coffea sp.
General Information
General Information
Commentary
Organism
metabolism
the enzyme catalyzes the first step in caffeine biosynthesis, pathway overview
Coffea sp.
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme catalyzes the first step in caffeine biosynthesis, pathway overview
Coffea sp.
Other publictions for EC 2.1.1.158
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
Synonyms
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)
721124
Mizuno
Essential region for 3-N methy ...
Coffea sp.
Z. Naturforsch. C
65c
257-265
2010
-
-
1
-
16
-
-
-
-
1
-
1
-
2
-
-
-
-
-
-
-
-
2
-
1
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
16
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-
696486
Kodama
Dimerization of N-methyltransf ...
Coffea arabica
Biochimie
90
547-551
2008
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700656
Deng
-
Fine control of caffeine biosy ...
Camellia sinensis
Phytochem. Lett.
1
195-198
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
667101
McCarthy
Cloning, expression, crystalli ...
Coffea canephora
Acta Crystallogr. Sect. F
F63
304-307
2007
-
-
1
1
-
-
-
-
-
-
2
1
-
1
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670598
McCarthy
The structure of two N-methylt ...
Coffea canephora
Plant Physiol.
144
879-889
2007
-
-
1
1
-
-
-
-
-
1
-
1
-
1
-
-
-
1
-
-
-
-
1
-
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
684218
McCarthy
Cloning, expression, crystalli ...
Coffea canephora
Acta Crystallogr. Sect. F
63
304-307
2007
-
-
1
1
-
-
-
-
-
-
2
-
-
1
-
-
1
-
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670621
Koshiro
-
Changes in content and biosynt ...
Coffea arabica, Coffea canephora
Plant Sci.
171
242-250
2006
-
-
-
-
-
-
-
-
-
-
-
6
-
2
-
-
-
-
-
8
-
-
9
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
-
8
-
-
9
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670568
Uefuji
Caffeine production in tobacco ...
Coffea arabica
Plant Mol. Biol.
59
221-227
2005
-
1
1
-
1
-
-
-
-
-
-
1
-
3
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670565
Ogita
Application of RNAi to confirm ...
Coffea arabica, Coffea canephora
Plant Mol. Biol.
54
931-941
2004
-
-
2
-
2
-
-
-
-
-
-
2
-
6
-
-
-
-
-
6
-
-
4
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
6
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
668586
Mizuno
The first committed step react ...
Coffea arabica
FEBS Lett.
547
56-60
2003
-
-
1
-
-
-
-
1
-
-
-
1
-
4
-
-
-
-
-
4
-
-
3
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
4
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
670578
Uefuji
Molecular cloning and function ...
Coffea arabica
Plant Physiol.
132
372-380
2003
-
-
1
-
-
-
-
1
-
-
2
1
-
1
-
-
-
-
-
3
1
-
1
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
2
1
-
-
-
-
-
3
1
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
655545
Koshiishi
A new caffeine biosynthetic pa ...
Camellia sinensis
FEBS Lett.
499
50-54
2001
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
1
1
-
2
-
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
669194
Ogawa
7-Methylxanthine methyltransfe ...
Coffea arabica
J. Biol. Chem.
276
8213-8218
2001
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
3
-
-
2
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
3
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667105
Moisyadi
-
Cloning and characterization o ...
Coffea arabica
Acta Hortic.
461
367-377
1998
-
-
1
-
-
-
-
-
-
-
1
1
-
1
-
-
1
-
-
-
-
-
2
1
2
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
670509
Waldhauser
Separation of the N-7 methyltr ...
Coffea arabica
Phytochemistry
45
1407-1414
1997
-
-
-
-
-
-
-
-
-
1
2
1
-
2
-
-
1
-
-
1
1
-
2
1
2
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
2
1
-
-
-
1
-
1
1
-
2
1
1
-
-
-
1
-
-
1
-
-
-
-
-
-
667116
Negishi
-
The role of xanthosine in the ...
Coffea arabica
Agric. Biol. Chem.
49
2221-2222
1985
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
1
-
-
5
-
-
2
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
5
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-