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Literature summary extracted from

  • Uefuji, H.; Ogita, S.; Yamaguchi, Y.; Koizumi, N.; Sano, H.
    Molecular cloning and functional characterization of three distinct N-methyltransferases involved in the caffeine biosynthetic pathway in coffee plants (2003), Plant Physiol., 132, 372-380.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.158 isozyme XMT1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged enzyme in Escherichia coli Bl21(DE3) Coffea arabica
2.1.1.159 isozyme MXMT1, DNA and amnio acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged isozymes in Escherichia coli Bl21(DE3) Coffea arabica
2.1.1.159 isozyme MXMT2, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged isozymes in Escherichia coli Bl21(DE3) Coffea arabica
2.1.1.160 isozyme DXMT1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the GST-tagged enzyme in Escherichia coli Bl21(DE3) Coffea arabica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.158 0.078
-
 Xanthosine recombinant XMT1 Coffea arabica
2.1.1.159 0.012
-
 S-adenosyl-L-methionine recombinant MXMT1 Coffea arabica
2.1.1.159 0.014
-
 S-adenosyl-L-methionine recombinant MXMT2 Coffea arabica
2.1.1.159 0.148
-
 7-methylxanthine recombinant MXMT1 Coffea arabica
2.1.1.159 0.251
-
 7-methylxanthine recombinant MXMT2 Coffea arabica
2.1.1.159 0.458
-
 paraxanthine recombinant MXMT1 Coffea arabica
2.1.1.159 0.738
-
 paraxanthine recombinant MXMT2 Coffea arabica
2.1.1.160 0.153
-
 S-adenosyl-L-methionine recombinant DXMT1 Coffea arabica
2.1.1.160 0.973
-
 paraxanthine recombinant DXMT1 Coffea arabica

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.158 41800
-
 2 * 41800, recombinant isozyme XMT1, SDS-PAGE Coffea arabica
2.1.1.158 69000
-
 recombinant isozyme XMT1, gel filtration Coffea arabica
2.1.1.159 43400
-
 2 * 43400, recombinant isozyme MXMT2, SDS-PAGE Coffea arabica
2.1.1.159 74000
-
 recombinant isozyme MXMT2, gel filtration Coffea arabica
2.1.1.160 43300
-
 2 * 43300, recombinant isozyme DXMT1, SDS-PAGE Coffea arabica
2.1.1.160 81000
-
 recombinant isozyme DXMT1, gel filtration Coffea arabica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.158 S-adenosyl-L-methionine + xanthosine Coffea arabica
-
 S-adenosyl-L-homocysteine + 7-methylxanthosine
-
 ir
2.1.1.159 S-adenosyl-L-methionine + 7-methylxanthine Coffea arabica
-
 S-adenosyl-L-homocysteine + 3,7-dimethylxanthine i.e. theobromine ir
2.1.1.159 S-adenosyl-L-methionine + paraxanthine Coffea arabica
-
 S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir
2.1.1.160 S-adenosyl-L-methionine + 3,7-dimethylxanthine Coffea arabica
-
 S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.158 Coffea arabica Q9AVK0 isozyme XMT1; isozyme XMT1
-
2.1.1.159 Coffea arabica Q84PP7 isozyme MXMT2; isozyme MXMT2
-
2.1.1.159 Coffea arabica Q9AVJ9 isozyme MXMT1; isozyme MXTM1
-
2.1.1.160 Coffea arabica Q8H0D2 isozyme DXMT1; isozyme DXMT1
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.158 flower bud high expression Coffea arabica
-
2.1.1.158 fruit high expression level in immature fruits, low expression in mature fruits Coffea arabica
-
2.1.1.158 leaf very high expression in young leaves, moderate expression in expanded leaves Coffea arabica
-
2.1.1.159 flower bud isozyme MXMT2 Coffea arabica
-
2.1.1.159 flower bud isozyme MXTM1 Coffea arabica
-
2.1.1.159 fruit immature, isozyme MXMT2 Coffea arabica
-
2.1.1.159 fruit immature, isozyme MXTM1 Coffea arabica
-
2.1.1.159 leaf high expression in young leaves of isozyme MXTM2, low expression level in expanded leaves Coffea arabica
-
2.1.1.159 leaf very high expression in young leaves of isozyme MXTM1, low expression level in expanded leaves Coffea arabica
-
2.1.1.160 flower bud low expression level Coffea arabica
-
2.1.1.160 fruit immature Coffea arabica
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.158 additional information
-
 recombinant enzyme Coffea arabica
2.1.1.159 additional information
-
 recombinant enzyme Coffea arabica
2.1.1.160 additional information
-
 recombinant enzyme Coffea arabica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.158 S-adenosyl-L-methionine + xanthosine
-
 Coffea arabica S-adenosyl-L-homocysteine + 7-methylxanthosine
-
 ir
2.1.1.159 S-adenosyl-L-methionine + 7-methylxanthine
-
 Coffea arabica S-adenosyl-L-homocysteine + 3,7-dimethylxanthine i.e. theobromine ir
2.1.1.159 S-adenosyl-L-methionine + 7-methylxanthine preferred substrate, 20fold higher activity by isozyme MXMT2 compared to activity with paraxanthine Coffea arabica S-adenosyl-L-homocysteine + 3,7-dimethylxanthine i.e. theobromine ir
2.1.1.159 S-adenosyl-L-methionine + 7-methylxanthine preferred substrate, 3fold higher activity by isozyme MXMT1 compared to activity with paraxanthine Coffea arabica S-adenosyl-L-homocysteine + 3,7-dimethylxanthine i.e. theobromine ir
2.1.1.159 S-adenosyl-L-methionine + paraxanthine
-
 Coffea arabica S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir
2.1.1.159 S-adenosyl-L-methionine + paraxanthine low activity by isozyme MXMT1 Coffea arabica S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir
2.1.1.159 S-adenosyl-L-methionine + paraxanthine low activity by isozyme MXMT2 Coffea arabica S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir
2.1.1.160 additional information the multifunctional isozyme DXMT1 catalyzes also the reaction theobromine synthase, EC 2.1.1.159, but at lower activity compared to its N1-methyltransferase, i.e. caffeine synthase activity Coffea arabica ?
-
 ?
2.1.1.160 S-adenosyl-L-methionine + 1,7-dimethylxanthine
-
 Coffea arabica S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine
-
 ?
2.1.1.160 S-adenosyl-L-methionine + 3,7-dimethylxanthine
-
 Coffea arabica S-adenosyl-L-homocysteine + 1,3,7-trimethylxanthine i.e. caffeine ir

Subunits

EC Number Subunits Comment Organism
2.1.1.158 dimer 2 * 41800, recombinant isozyme XMT1, SDS-PAGE Coffea arabica
2.1.1.159 dimer 2 * 43400, recombinant isozyme MXMT2, SDS-PAGE Coffea arabica
2.1.1.160 dimer 2 * 43300, recombinant isozyme DXMT1, SDS-PAGE Coffea arabica

Synonyms

EC Number Synonyms Comment Organism
2.1.1.158 XMT1
-
 Coffea arabica
2.1.1.159 MXMT1
-
 Coffea arabica
2.1.1.159 MXMT2
-
 Coffea arabica
2.1.1.160 DXMT1
-
 Coffea arabica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.158 27
-
 assay at Coffea arabica
2.1.1.159 27
-
 assay at Coffea arabica
2.1.1.160 27
-
 assay at Coffea arabica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.158 8
-
 assay at Coffea arabica
2.1.1.159 8
-
 assay at Coffea arabica
2.1.1.160 8
-
 assay at Coffea arabica