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Literature summary extracted from

  • De Luca, V.; Ibrahim, R.K.
    Enzymatic synthesis of polymethylated flavonols in Chrysosplenium americanum. I. Partial purification and some properties of S-adenosyl-L-methionine:flavonol 3-, 6-, 7-, and 4-O-methyltransferases (1985), Arch. Biochem. Biophys., 238, 596-605.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.1.1.76 2-mercaptoethanol relative activity 105% Chrysosplenium americanum
2.1.1.76 iodoacetamide relative activity 105% Chrysosplenium americanum

General Stability

EC Number General Stability Organism
2.1.1.82 loss of activity during chromatofocusing Chrysosplenium americanum

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.76 N-ethylmaleimide
-
Chrysosplenium americanum
2.1.1.76 p-chloromercuribenzoate
-
Chrysosplenium americanum
2.1.1.82 N-ethylmaleimide
-
Chrysosplenium americanum
2.1.1.82 p-chloromercuribenzoate
-
Chrysosplenium americanum
2.1.1.83 N-ethylmaleimide
-
Chrysosplenium americanum
2.1.1.83 p-chloromercuribenzoate
-
Chrysosplenium americanum
2.1.1.84 EDTA
-
Chrysosplenium americanum
2.1.1.84 N-ethylmaleimide
-
Chrysosplenium americanum
2.1.1.84 p-chloromercuribenzoate
-
Chrysosplenium americanum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.76 K+ relative activity 115% Chrysosplenium americanum
2.1.1.76 Mn2+ relative activity 110% Chrysosplenium americanum
2.1.1.84 Mg2+ activates Chrysosplenium americanum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.76 57000
-
gel filtration Chrysosplenium americanum
2.1.1.76 65000
-
gel filtration Chrysosplenium americanum
2.1.1.82 57000
-
gel filtration Chrysosplenium americanum
2.1.1.83 57000
-
gel filtration Chrysosplenium americanum
2.1.1.84 57000
-
gel filtration Chrysosplenium americanum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.76 S-adenosyl-L-methionine + 3,5,7,3',4'-pentahydroxyflavone Citrofortunella microcarpa
-
S-adenosyl-L-homocysteine + 3-methoxy-5,7,3',4'-tetrahydroxyflavone
-
?
2.1.1.76 S-adenosyl-L-methionine + 3,5,7,3',4'-pentahydroxyflavone Chrysosplenium americanum
-
S-adenosyl-L-homocysteine + 3-methoxy-5,7,3',4'-tetrahydroxyflavone 3-O-methylquercetin ?
2.1.1.82 S-adenosyl-L-methionine + 3',4',5,7-tetrahydroxy-3-methoxyflavone Chrysosplenium americanum involved with EC 2.1.1.76 and 2.1.1.83 in the methylation of quercetin S-adenosyl-L-homocysteine + 3',4',5-trihydroxy-3,7-dimethoxyflavone trivial name 3,7-dimethylquercetin ?
2.1.1.83 S-adenosyl-L-methionine + 3',4',5-trihydroxy-3,7-dimethoxyflavone Chrysosplenium americanum trivial name 3,7-dimethylquercetin, enzyme does not act on flavones, dihydroflavonols or their glucosides, involved with EC 2.1.1.76 and 2.1.1.82 in the methylation of quercitin to 3,7,4'-trimethylquercitin S-adenosyl-L-homocysteine + 3',5-dihydroxy-3,4',7-trimethoxyflavone trivial name 3,7,4'-trimethylquercetin ?
2.1.1.84 S-adenosyl-L-methionine + 3',4',5,6-tetrahydroxy-3,7-dimethoxyflavone Chrysosplenium americanum
-
S-adenosyl-L-homocysteine + 3',4',5-trihydroxy-3,6,7-trimethoxyflavone
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.76 Chrysosplenium americanum
-
-
-
2.1.1.76 Citrofortunella microcarpa
-
calamondin orange, synonym Citrofortunella mitis
-
2.1.1.82 Chrysosplenium americanum
-
-
-
2.1.1.83 Chrysosplenium americanum
-
-
-
2.1.1.84 Chrysosplenium americanum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.76 partially Chrysosplenium americanum
2.1.1.76 partially Citrofortunella microcarpa
2.1.1.82 loss of activity during chromatofocusing possibly due to instability of the enzyme in the purified form or to the loss of some factors required for activity Chrysosplenium americanum
2.1.1.83
-
Chrysosplenium americanum
2.1.1.84 ammonium sulfate, Sephacryl S-200, hydroxylapatite, chromatofocusing Chrysosplenium americanum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.82 leaf
-
Chrysosplenium americanum
-
2.1.1.83 leaf
-
Chrysosplenium americanum
-
2.1.1.84 leaf
-
Chrysosplenium americanum
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.76 additional information
-
53 pkat/mg Chrysosplenium americanum
2.1.1.83 7.2
-
-
Chrysosplenium americanum
2.1.1.84 2.64
-
-
Chrysosplenium americanum

Storage Stability

EC Number Storage Stability Organism
2.1.1.76 -15°C, purified enzyme, 0.2 M potassium phosphate, pH 8.0, 20% glycerol, 5 mM dithiothreitol, stable for several months Chrysosplenium americanum
2.1.1.76 4°C, partially purified enzyme loses 80% of its activity over a period of 4-6 weeks Chrysosplenium americanum
2.1.1.82 -15°C, 200 mM potassium phosphate buffer, pH 8, 20% v/v glycerol, 5 mM dithiothreitol, several months, no loss of activity Chrysosplenium americanum
2.1.1.82 4°C, 4-6 weeks, 80% loss of activity Chrysosplenium americanum
2.1.1.83 -15°C, 200 mM potassium phosphate buffer, pH 8, 20% glycerol, 5 mM dithiothreitol, several months, no loss of activity Chrysosplenium americanum
2.1.1.83 4°C, 4-6 weeks, 80% loss of activity Chrysosplenium americanum
2.1.1.84 -15°C, 0.2 M potassium phosphate, pH 8, 20% glycerol, 5 mM dithiothreitol, for several months, no loss of activity Chrysosplenium americanum
2.1.1.84 4°C, 4-6 weeks, 80% loss of activity Chrysosplenium americanum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.76 S-adenosyl-L-methionine + 3,5,7,3',4'-pentahydroxyflavone
-
Chrysosplenium americanum S-adenosyl-L-homocysteine + 3-methoxy-5,7,3',4'-tetrahydroxyflavone
-
?
2.1.1.76 S-adenosyl-L-methionine + 3,5,7,3',4'-pentahydroxyflavone
-
Citrofortunella microcarpa S-adenosyl-L-homocysteine + 3-methoxy-5,7,3',4'-tetrahydroxyflavone
-
?
2.1.1.76 S-adenosyl-L-methionine + 3,5,7,3',4'-pentahydroxyflavone
-
Chrysosplenium americanum S-adenosyl-L-homocysteine + 3-methoxy-5,7,3',4'-tetrahydroxyflavone 3-O-methylquercetin ?
2.1.1.76 S-adenosyl-L-methionine + 7-methylquercetin i.e. rhamnetin, 30% of the activity with quercetin Chrysosplenium americanum ?
-
?
2.1.1.82 S-adenosyl-L-methionine + 3',4',5,7-tetrahydroxy-3-methoxyflavone trivial name 3-methylquercetin, specific for 3-methylquercetin Chrysosplenium americanum S-adenosyl-L-homocysteine + 3',4',5-trihydroxy-3,7-dimethoxyflavone trivial name 3,7-dimethylquercetin ?
2.1.1.82 S-adenosyl-L-methionine + 3',4',5,7-tetrahydroxy-3-methoxyflavone involved with EC 2.1.1.76 and 2.1.1.83 in the methylation of quercetin Chrysosplenium americanum S-adenosyl-L-homocysteine + 3',4',5-trihydroxy-3,7-dimethoxyflavone trivial name 3,7-dimethylquercetin ?
2.1.1.83 S-adenosyl-L-methionine + 3',4',5-trihydroxy-3,7-dimethoxyflavone trivial name 3,7-dimethylquercetin, enzyme does not act on flavones, dihydroflavonols or their glucosides Chrysosplenium americanum S-adenosyl-L-homocysteine + 3',5-dihydroxy-3,4',7-trimethoxyflavone trivial name 3,7,4'-trimethylquercetin ?
2.1.1.83 S-adenosyl-L-methionine + 3',4',5-trihydroxy-3,7-dimethoxyflavone trivial name 3,7-dimethylquercetin, enzyme does not act on flavones, dihydroflavonols or their glucosides, involved with EC 2.1.1.76 and 2.1.1.82 in the methylation of quercitin to 3,7,4'-trimethylquercitin Chrysosplenium americanum S-adenosyl-L-homocysteine + 3',5-dihydroxy-3,4',7-trimethoxyflavone trivial name 3,7,4'-trimethylquercetin ?
2.1.1.83 S-adenosyl-L-methionine + 3,7-dimethylquercetagetin
-
Chrysosplenium americanum S-adenosyl-L-homocysteine + 3,7,4'-trimethylquercetagetin + H+
-
?
2.1.1.84 3,7,3'-trimethylquercetagetin + S-adenosyl-L-methionine
-
Chrysosplenium americanum 3,6,7',3'-tetrametylquercetagetin + S-adenosyl-L-homocysteine
-
?
2.1.1.84 8-hydroxykaempferol + S-adenosyl-L-methionine 25% of activity with 3',4',5,6-tetrahydroxy-3,7-dimethoxyflavone Chrysosplenium americanum ?
-
?
2.1.1.84 S-adenosyl-L-methionine + 3',4',5,6-tetrahydroxy-3,7-dimethoxyflavone
-
Chrysosplenium americanum S-adenosyl-L-homocysteine + 3',4',5-trihydroxy-3,6,7-trimethoxyflavone
-
?

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.76 7.8 8.2
-
Chrysosplenium americanum
2.1.1.82 7.8 8.2
-
Chrysosplenium americanum
2.1.1.83 8.8 9
-
Chrysosplenium americanum
2.1.1.84 8.8 9
-
Chrysosplenium americanum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.76 7.2 9.3 about 50% of activity maximum at pH 7.2 and 9.3 Chrysosplenium americanum
2.1.1.82 7.2 9.5 approx. 50% of maximal activity at pH 7.2 and 9.5 Chrysosplenium americanum
2.1.1.83 7.2 9.5 approx. 50% of maximal activity at pH 7.2 and 9.5 Chrysosplenium americanum