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Literature summary extracted from

  • Wu, X.; Yuwen, M.; Pu, Z.; Zhao, Z.; Yu, H.; Zha, J.
    Engineering of flavonoid 3-O-methyltransferase for improved biosynthesis of chrysoeriol in Escherichia coli (2023), Appl. Microbiol. Biotechnol., 107, 1663-1672.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.267 expressed in Escherichia coli BL21(DE3) cells synthetic construct

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.42 A372C complete loss of activity Oryza sativa Japonica Group
2.1.1.42 A372K complete loss of activity Oryza sativa Japonica Group
2.1.1.42 A372L complete loss of activity Oryza sativa Japonica Group
2.1.1.42 A372Q complete loss of activity Oryza sativa Japonica Group
2.1.1.42 C316F complete loss of activity Oryza sativa Japonica Group
2.1.1.42 G330R complete loss of activity Oryza sativa Japonica Group
2.1.1.42 L34P complete loss of activity Oryza sativa Japonica Group
2.1.1.42 L34Q mutant shows improved production of chrysoeriol from luteolin due to a higher enzyme expression level Oryza sativa Japonica Group
2.1.1.42 N373C complete loss of activity Oryza sativa Japonica Group
2.1.1.42 N373D complete loss of activity Oryza sativa Japonica Group
2.1.1.42 N373K complete loss of activity Oryza sativa Japonica Group
2.1.1.42 N373T complete loss of activity Oryza sativa Japonica Group
2.1.1.42 N373W complete loss of activity Oryza sativa Japonica Group
2.1.1.42 W284A mutant shows improved production of chrysoeriol from luteolin due to higher efficiency in substrate binding and catalysis Oryza sativa Japonica Group
2.1.1.42 W284P complete loss of activity Oryza sativa Japonica Group
2.1.1.267 A372C inactive synthetic construct
2.1.1.267 A372K inactive synthetic construct
2.1.1.267 A372L inactive synthetic construct
2.1.1.267 A372Q inactive synthetic construct
2.1.1.267 C316F inactive synthetic construct
2.1.1.267 G330R inactive synthetic construct
2.1.1.267 L34P inactive synthetic construct
2.1.1.267 L34Q the mutant shows 88% increased chrysoeriol biosynthesis activity and higher substrate affinity compared to the wild type enzyme synthetic construct
2.1.1.267 N373C inactive synthetic construct
2.1.1.267 N373D inactive synthetic construct
2.1.1.267 N373K inactive synthetic construct
2.1.1.267 N373T inactive synthetic construct
2.1.1.267 N373W inactive synthetic construct
2.1.1.267 W284A the mutant shows 75% increased chrysoeriol biosynthesis activity and higher expression level compared to the wild type enzyme synthetic construct
2.1.1.267 W284P inactive synthetic construct

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.42 0.00358
-
 S-adenosyl-L-methionine mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.00413
-
 luteolin mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.02077
-
 luteolin mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.02136
-
 S-adenosyl-L-methionine mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.0277
-
 luteolin wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.03541
-
 S-adenosyl-L-methionine wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.267 0.00358
-
 S-adenosyl-L-methionine mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.00413
-
 luteolin mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.02077
-
 luteolin mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.02136
-
 S-adenosyl-L-methionine mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.0277
-
 luteolin wild type enzyme, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.03541
-
 S-adenosyl-L-methionine wild type enzyme, at pH 8.0 and 37°C synthetic construct

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.42 Oryza sativa Japonica Group Q6ZD89
-
-
2.1.1.267 synthetic construct OP326931
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.267 Ni-NTA agarose resin column chromatography synthetic construct

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.42 S-adenosyl-L-methionine + luteolin
-
 Oryza sativa Japonica Group S-adenosyl-L-homocysteine + chrysoeriol
-
 ?
2.1.1.267 S-adenosyl-L-methionine + luteolin
-
 synthetic construct S-adenosyl-L-homocysteine + chrysoeriol
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.267 ? x * 41200, His-tagged enzyme, SDS-PAGE synthetic construct

Synonyms

EC Number Synonyms Comment Organism
2.1.1.42 COMT
-
 Oryza sativa Japonica Group
2.1.1.42 ROMT-9
-
 Oryza sativa Japonica Group
2.1.1.267 3'-OMT
-
 synthetic construct
2.1.1.267 ROMT-9
-
 synthetic construct

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.42 0.07
-
 luteolin mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.07
-
 S-adenosyl-L-methionine mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.25
-
 S-adenosyl-L-methionine mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.27
-
 luteolin mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.41
-
 S-adenosyl-L-methionine wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 0.51
-
 luteolin wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.267 0.07
-
 luteolin mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.07
-
 S-adenosyl-L-methionine mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.25
-
 S-adenosyl-L-methionine mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.27
-
 luteolin mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.41
-
 S-adenosyl-L-methionine wild type enzyme, at pH 8.0 and 37°C synthetic construct
2.1.1.267 0.51
-
 luteolin wild type enzyme, at pH 8.0 and 37°C synthetic construct

Cofactor

EC Number Cofactor Comment Organism Structure
2.1.1.267 S-adenosyl-L-methionine
-
 synthetic construct

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.42 11.44
-
 S-adenosyl-L-methionine wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 11.68
-
 S-adenosyl-L-methionine mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 13.03
-
 luteolin mutant L34Q, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 17.84
-
 luteolin mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 18.58
-
 luteolin wild-type, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.42 19.22
-
 S-adenosyl-L-methionine mutant W284A, pH 8, 37°C Oryza sativa Japonica Group
2.1.1.267 11.44
-
 S-adenosyl-L-methionine wild type enzyme, at pH 8.0 and 37°C synthetic construct
2.1.1.267 11.68
-
 S-adenosyl-L-methionine mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 13.03
-
 luteolin mutant enzyme L34Q, at pH 8.0 and 37°C synthetic construct
2.1.1.267 17.84
-
 luteolin mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct
2.1.1.267 18.58
-
 luteolin wild type enzyme, at pH 8.0 and 37°C synthetic construct
2.1.1.267 19.22
-
 S-adenosyl-L-methionine mutant enzyme W284A, at pH 8.0 and 37°C synthetic construct