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Literature summary extracted from

  • Itoh, N.; Iwata, C.; Toda, H.
    Molecular cloning and characterization of a flavonoid-O-methyltransferase with broad substrate specificity and regioselectivity from Citrus depressa (2016), BMC Plant Biol., 16, 180 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.42 expression in Escherichia coli Citrus depressa
2.1.1.76 gene FOMT5, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of soluble homodimeric His6-tagged enzyme in Escherichia coli strain BL21(DE3) Citrus depressa
2.1.1.267 gene FOMT1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis Citrus depressa
2.1.1.267 gene FOMT3, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis Citrus depressa
2.1.1.267 gene FOMT4, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis Citrus depressa
2.1.1.267 gene FOMT5, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of soluble homodimeric His6-tagged enzyme in Escherichia coli strain BL21(DE3) Citrus depressa
2.1.1.267 gene FOMT6, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis Citrus depressa

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.1.1.42 soluble
-
 Citrus depressa
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.76 additional information the enzyme is an Mg2+-independent O-methyltransferase Citrus depressa
2.1.1.267 additional information the enzyme is an Mg2+-independent O-methyltransferase Citrus depressa

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.42 88000
-
 gel filtration Citrus depressa
2.1.1.76 88000
-
 recombinant His6-tagged enzyme, gel filtration Citrus depressa
2.1.1.267 88000
-
 recombinant His6-tagged enzyme, gel filtration Citrus depressa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.76 4 S-adenosyl-L-methionine + quercetin Citrus depressa whole-cell biocatalysis using CdFOMT5 expressed in Escherichia coli cells is performed using quercetin as a substrate, and 3,3',5,7-tetramethylated quercetin is obtained as the final product 4 S-adenosyl-L-homocysteine + 3,3',5,7-tetramethoxyquercetin
-
 ?
2.1.1.267 4 S-adenosyl-L-methionine + quercetin Citrus depressa whole-cell biocatalysis using CdFOMT5 expressed in Escherichia coli cells is performed using quercetin as a substrate, and 3,3',5,7-tetramethylated quercetin is obtained as the final product 4 S-adenosyl-L-homocysteine + 3,3',5,7-tetramethoxyquercetin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.42 Citrus depressa A0A125T1T5
-
-
2.1.1.76 Citrus depressa A0A125T1T5
-
-
2.1.1.267 Citrus depressa A0A120MXL9
-
-
2.1.1.267 Citrus depressa A0A120MXM2
-
-
2.1.1.267 Citrus depressa A0A120MXM3
-
-
2.1.1.267 Citrus depressa A0A125T1T4
-
-
2.1.1.267 Citrus depressa A0A125T1T5
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.42 fruit
-
 Citrus depressa
-
2.1.1.76 fruit
-
 Citrus depressa
-
2.1.1.76 pericarp
-
 Citrus depressa
-
2.1.1.267 fruit
-
 Citrus depressa
-
2.1.1.267 pericarp
-
 Citrus depressa
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.42 4 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa 4 S-adenosyl-L-homocysteine + 3,3',5,7-tetra-O-methyl-quercetin
-
 ?
2.1.1.42 additional information enzyme shows methylation activity for the 3-, 5-, 6-, and 7-hydroxyl groups of flavones. The highest activity is observed for 3-hydroxyflavone (flavonol), followed by 7-hydroxyflavone with a relative activity of 15.6% of that of 3-hydroxyflavone and 5-hydroxyflavone with that of 13.5%. Very weak activity is observed for 6-hydroxyflavone and no activity for 3'- or 4'-hydroxyflavone and 7-methoxy-8-hydroxyflavone. FOMT5 does not exhibit a sequential methylation order for 3-, 5-, and 7-hydroxyl groups of quercetin. No detection of polymethylated products from naringenin, (-)-epicatechin, and equol, and cyanidin is not a substrate Citrus depressa ?
-
-
2.1.1.42 S-adenosyl-L-methionine + (-)-epicatechin
-
 Citrus depressa S-adenosyl-L-homocysteine + ?
-
 ?
2.1.1.42 S-adenosyl-L-methionine + (2S)-naringenin
-
 Citrus depressa S-adenosyl-L-homocysteine + ?
-
 ?
2.1.1.42 S-adenosyl-L-methionine + equol
-
 Citrus depressa S-adenosyl-L-homocysteine + ?
-
 ?
2.1.1.76 4 S-adenosyl-L-methionine + quercetin whole-cell biocatalysis using CdFOMT5 expressed in Escherichia coli cells is performed using quercetin as a substrate, and 3,3',5,7-tetramethylated quercetin is obtained as the final product Citrus depressa 4 S-adenosyl-L-homocysteine + 3,3',5,7-tetramethoxyquercetin
-
 ?
2.1.1.76 additional information the flavonoid-O-methyltransferase from Citrus depressa has a broad substrate specificity and regioselectivity. Isozyme CdFOMT5 exhibits O-methyltransferase activity for quercetin, naringenin, (-)-epicatechin, and equol using S-adenosyl-L-methionine (SAM) as a methyl donor. The recombinant CdFOMT5 CdFOMT5 can catalyze the O-methylation of at least three hydroxyl groups of quercetin, and di- or tri-O-methylated quercetin products are obtained by this enzymatic reaction. CdFMOT5 prefers flavonol (3-hydroxyflavone) to other flavonoid structures. Thus, substrate structure, especially the C-ring in flavonoids, may strongly affect the substrate preference, including regioselectivity, of CdFOMT5 Citrus depressa ?
-
-
2.1.1.76 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 3'-methoxyquercetin
-
 ?
2.1.1.76 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 3-methoxyquercetin
-
 ?
2.1.1.76 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 5-methoxyquercetin
-
 ?
2.1.1.76 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 7-methoxyquercetin
-
 ?
2.1.1.267 4 S-adenosyl-L-methionine + quercetin whole-cell biocatalysis using CdFOMT5 expressed in Escherichia coli cells is performed using quercetin as a substrate, and 3,3',5,7-tetramethylated quercetin is obtained as the final product Citrus depressa 4 S-adenosyl-L-homocysteine + 3,3',5,7-tetramethoxyquercetin
-
 ?
2.1.1.267 additional information the flavonoid-O-methyltransferase from Citrus depressa has a broad substrate specificity and regioselectivity Citrus depressa ?
-
-
2.1.1.267 additional information the flavonoid-O-methyltransferase from Citrus depressa has a broad substrate specificity and regioselectivity. Isozyme CdFOMT5 exhibits O-methyltransferase activity for quercetin, naringenin, (-)-epicatechin, and equol using S-adenosyl-L-methionine (SAM) as a methyl donor. The recombinant CdFOMT5 CdFOMT5 can catalyze the O-methylation of at least three hydroxyl groups of quercetin, and di- or tri-O-methylated quercetin products are obtained by this enzymatic reaction. CdFMOT5 prefers flavonol (3-hydroxyflavone) to other flavonoid structures. Thus, substrate structure, especially the C-ring in flavonoids, may strongly affect the substrate preference, including regioselectivity, of CdFOMT5 Citrus depressa ?
-
-
2.1.1.267 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 3'-methoxyquercetin
-
 ?
2.1.1.267 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 3-methoxyquercetin
-
 ?
2.1.1.267 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 5-methoxyquercetin
-
 ?
2.1.1.267 S-adenosyl-L-methionine + quercetin
-
 Citrus depressa S-adenosyl-L-homocysteine + 7-methoxyquercetin
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.42 ? x * 42000, SDS-PAGE, recombinant protein including His-tag Citrus depressa
2.1.1.76 homodimer 2 * 42030, recombinant His6-tagged enzyme, sequence calculation, 2 * 42000, recombinant His6-tagged enzyme, SDS-PAGE Citrus depressa
2.1.1.267 homodimer 2 * 42030, recombinant His6-tagged enzyme, sequence calculation, 2 * 42000, recombinant His6-tagged enzyme, SDS-PAGE Citrus depressa

Synonyms

EC Number Synonyms Comment Organism
2.1.1.42 FOMT5
-
 Citrus depressa
2.1.1.76 CdFOMT5
-
 Citrus depressa
2.1.1.76 flavonoid-O-methyltransferase
-
 Citrus depressa
2.1.1.76 FOMT5
-
 Citrus depressa
2.1.1.76 Mg2+-independent O-methyltransferase
-
 Citrus depressa
2.1.1.76 More see also EC 2.1.1.267 Citrus depressa
2.1.1.267 CdFOMT1
-
 Citrus depressa
2.1.1.267 CdFOMT3
-
 Citrus depressa
2.1.1.267 CdFOMT4
-
 Citrus depressa
2.1.1.267 CdFOMT5
-
 Citrus depressa
2.1.1.267 CdFOMT6
-
 Citrus depressa
2.1.1.267 flavonoid-O-methyltransferase
-
 Citrus depressa
2.1.1.267 FOMT1
-
 Citrus depressa
2.1.1.267 FOMT3
-
 Citrus depressa
2.1.1.267 FOMT4
-
 Citrus depressa
2.1.1.267 FOMT5
-
 Citrus depressa
2.1.1.267 FOMT6
-
 Citrus depressa
2.1.1.267 Mg2+-independent O-methyltransferase
-
 Citrus depressa
2.1.1.267 More see also EC 2.1.1.76 Citrus depressa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.42 45
-
-
 Citrus depressa
2.1.1.76 45
-
 recombinant enzyme Citrus depressa
2.1.1.267 45
-
 recombinant enzyme Citrus depressa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.42 7
-
-
 Citrus depressa
2.1.1.76 7
-
 recombinant enzyme Citrus depressa
2.1.1.267 7
-
 recombinant enzyme Citrus depressa

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.76 5.5 9 recombinant enzyme, over 50% activity within this range Citrus depressa
2.1.1.267 5.5 9 recombinant enzyme, over 50% activity within this range Citrus depressa

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.76 S-adenosyl-L-methionine
-
 Citrus depressa
2.1.1.267 S-adenosyl-L-methionine
-
 Citrus depressa

pI Value

EC Number Organism Comment pI Value Maximum pI Value
2.1.1.42 Citrus depressa calculated from sequence
-
 5.8
2.1.1.76 Citrus depressa recombinant detagged enzyme, sequence calculation
-
 5.79
2.1.1.267 Citrus depressa recombinant detagged enzyme, sequence calculation
-
 5.79