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Literature summary extracted from

  • Joe, E.J.; Kim, B.G.; An, B.C.; Chong, Y.; Ahn, J.H.
    Engineering of flavonoid O-methyltransferase for a novel regioselectivity (2010), Mol. Cells, 30, 137-141.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.B75 gene POMT7, error-prone polymerase chain reaction, expression of GST-tagged wild-type POMT7 and mutant POMT7-M1 in Escherichia coli Populus sp.
2.1.1.B76
-
Populus sp.
2.1.1.B76 gene POMT7, error-prone polymerase chain reaction, expression of GST-tagged wild-type POMT7 and mutant POMT7-M1 in Escherichia coli Populus sp.
2.1.1.82 expressed in Escherichia coli BL21 cells Populus sp.

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.B75 D257G random mutagenesis, the mutant of POMT-7, POMT7-M1, has a novel regioselectivity, from large scale mutant screening, mutant D257G methylates the 3-hydroxyl group of flavonols in addition to 7-hydrdoxyl group Populus sp.
2.1.1.B76 D257G random mutagenesis, the mutant of POMT-7, POMT7-M1, has a novel regioselectivity, from large scale mutant screening, mutant D257G methylates the 3-hydroxyl group of flavonols in addition to 7-hydrdoxyl group Populus sp.
2.1.1.B76 N257G the mutant methylates the 3-hydroxyl group of flavonols in addition to 7-hydrdoxyl group. The wild type POMT7 exhibits better activity toward kaempferol and quercetin than the mutant enzyme Populus sp.
2.1.1.82 D257G the mutant methylates the 3-hydroxyl group of flavonols in addition to the 7-hydrdoxyl group Populus sp.
2.1.1.82 L101M the mutant shows wild type reactivity with kaempferol and quercetin Populus sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.B75 0.0298
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B75 0.0324
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B75 0.0324
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B75 0.0378
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 0.0298
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 0.0298
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 0.0324
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 0.0324
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 0.0324
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 0.0324
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 0.0378
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 0.0378
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.82 0.0298
-
quercetin wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.0324
-
quercetin mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.0324
-
kaempferol wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.0378
-
kaempferol mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.82 60000
-
x * 60000, SDS-PAGE Populus sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.B75 S-adenosyl-L-methionine + kaempferol Populus sp.
-
S-adenosyl-L-homocysteine + 7-O-methylkaempferol
-
?
2.1.1.B75 S-adenosyl-L-methionine + quercetin Populus sp.
-
S-adenosyl-L-homocysteine + 7-O-methylquercetin
-
?
2.1.1.B76 S-adenosyl-L-methionine + kaempferol Populus sp.
-
S-adenosyl-L-homocysteine + 7-O-methylkaempferol
-
?
2.1.1.B76 S-adenosyl-L-methionine + quercetin Populus sp.
-
S-adenosyl-L-homocysteine + 7-O-methylquercetin
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.B75 Populus sp.
-
-
-
2.1.1.B76 Populus sp.
-
-
-
2.1.1.82 Populus sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.B75 recombinant GST-tagged wild-type POMT7 and mutant POMT7-M1 fromEscherichia coli by glutathione affinity chromatography Populus sp.
2.1.1.B76 recombinant GST-tagged wild-type POMT7 and mutant POMT7-M1 from Escherichia coli by glutathione affinity chromatography Populus sp.
2.1.1.B76 recombinent POMT7 and mutants Populus sp.
2.1.1.82 GST affinity column chromatography Populus sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.B75 2 S-adenosyl-L-methionine + kaempferol activity only of enzyme mutant POMT7-M1, D257G, not of wild-type POMT7 Populus sp. 2 S-adenosyl-L-homocysteine + 3,7-O-dimethylkaempferol
-
?
2.1.1.B75 2 S-adenosyl-L-methionine + quercetin activity only of enzyme mutant POMT7-M1, D257G, not of wild-type POMT7, molecular docking of quercetin into the modeled structure of Asp257Gly Populus sp. 2 S-adenosyl-L-homocysteine + 3,7-O-dimethylquercetin
-
?
2.1.1.B75 S-adenosyl-L-methionine + kaempferol
-
Populus sp. S-adenosyl-L-homocysteine + 7-O-methylkaempferol
-
?
2.1.1.B75 S-adenosyl-L-methionine + quercetin
-
Populus sp. S-adenosyl-L-homocysteine + 7-O-methylquercetin
-
?
2.1.1.B76 2 S-adenosyl-L-methionine + kaempferol activity only of enzyme mutant POMT7-M1, D257G, not of wild-type POMT7 Populus sp. S-adenosyl-L-homocysteine + 3,7-O-dimethylkaempferol
-
?
2.1.1.B76 2 S-adenosyl-L-methionine + quercetin activity only of enzyme mutant POMT7-M1, D257G, not of wild-type POMT7, molecular docking of quercetin into the modeled structure of Asp257Gly Populus sp. S-adenosyl-L-homocysteine + 3,7-O-dimethylquercetin
-
?
2.1.1.B76 S-adenosyl-L-methionine + 3,3',4',5,7-pentahydroxyflavone i.e. quercetin Populus sp. S-adenosyl-L-homocysteine + 3,3',4',5-tetrahydroxy-7-methoxyflavone
-
?
2.1.1.B76 S-adenosyl-L-methionine + 3,4',5,7-tetrahydroxyflavanone i.e. kaempferol Populus sp. S-adenosyl-L-homocysteine + 3,4',5-trihydroxy-7-methoxyflavanone
-
?
2.1.1.B76 S-adenosyl-L-methionine + kaempferol
-
Populus sp. S-adenosyl-L-homocysteine + 7-O-methylkaempferol
-
?
2.1.1.B76 S-adenosyl-L-methionine + quercetin
-
Populus sp. S-adenosyl-L-homocysteine + 7-O-methylquercetin
-
?
2.1.1.82 S-adenosyl-L-methionine + kaempferol
-
Populus sp. 7-O-methylkaempferol
-
?
2.1.1.82 S-adenosyl-L-methionine + quercetin
-
Populus sp. 7-O-methylquercetin
-
?

Subunits

EC Number Subunits Comment Organism
2.1.1.82 ? x * 60000, SDS-PAGE Populus sp.

Synonyms

EC Number Synonyms Comment Organism
2.1.1.B75 flavonoid O-methyltransferase
-
Populus sp.
2.1.1.B75 POMT7
-
Populus sp.
2.1.1.B76 flavonoid O-methyltransferase
-
Populus sp.
2.1.1.B76 POMT7
-
Populus sp.
2.1.1.82 flavone 7-O-methyltransferase
-
Populus sp.
2.1.1.82 MT7
-
Populus sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.B75 37
-
assay at Populus sp.
2.1.1.B76 37
-
assay at Populus sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.B75 0.064
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B75 0.064
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B75 0.07
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B75 0.071
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 0.064
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 0.064
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 0.064
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 0.064
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 0.07
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 0.07
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 0.071
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 0.071
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.82 0.064
-
quercetin mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.064
-
kaempferol wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.07
-
kaempferol mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 0.071
-
quercetin wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.B75 7.5
-
assay at Populus sp.
2.1.1.B76 7.5
-
assay at Populus sp.

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.B75 S-adenosyl-L-methionine
-
Populus sp.
2.1.1.B76 S-adenosyl-L-methionine
-
Populus sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.B75 1.7
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B75 1.9
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B75 1.9
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B75 2.4
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 1.7
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 1.7
-
quercetin pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 1.9
-
3,4',5,7-tetrahydroxyflavanone i.e. kaempferol, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 1.9
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, mutant enzyme N257G Populus sp.
2.1.1.B76 1.9
-
kaempferol pH 7.5, 37°C, recombinant mutant POMT7-M1 Populus sp.
2.1.1.B76 1.9
-
quercetin pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.B76 2.4
-
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-4H-1-benzopyran-4-one i.e. quercetin, pH 7.5, 37°C, wild-type enzyme Populus sp.
2.1.1.B76 2.4
-
kaempferol pH 7.5, 37°C, recombinant wild-type POMT7 Populus sp.
2.1.1.82 17
-
kaempferol mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 19
-
quercetin mutant enzyme D257G, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 19
-
kaempferol wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.
2.1.1.82 24
-
quercetin wild type enzyme, in 10 mM Tris/HCl buffer (pH 7.5), at 37°C Populus sp.