Cloned (Comment) | Organism |
---|---|
recombinant expression in Escherichia coli strain BL21(DE3) | Clarkia breweri |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0068 | - |
salicylate | pH 7.5, 30°C | Clarkia breweri |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
KCl | activates at 1 mM | Clarkia breweri |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + salicylate | Clarkia breweri | - |
methyl salicylate + S-adenosyl-L-homocysteine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clarkia breweri | Q9SPV4 | i.e. Eucharidium breweri | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | development and evaluation of an enzyme-coupled assay for monitoring methyltransferase activity, overview. Since S-adenosyl-L-homocysteine is a key by-product of reactions catalyzed by S-adenosyl methionine-dependent methyltransferases, the coupling enzymes are used to assess the activities of EcoRI methyltransferase and a salicylic acid methyltransferase from Clarkia breweri in the presence of S-adenosyl methionine. In the case of the salicylic acid methyltransferase, detectable activity is observed for several substrates including salicylic acid, benzoic acid, 3-hydroxybenzoic acid, and vanillic acid, substrate specificity, overview. Additionally, the de novo synthesis of the relatively expensive and unstable cosubstrate, S-adenosyl methionine, catalyzed by methionine adenosyltransferase can be incorporated within the assay. The assay offers a high level of sensitivity that permits continuous and reliable monitoring of methyltransferase activities. The assay enzymes, 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (Mtn), xanthine oxidase (XOD), and horse radish peroxidase (HRP), are able to operate in a tandem manner to generate a fluorescence signal in the presence of SAH, the key by-product of reactions catalyzed by SAM-dependent methyltransferases. Poor or no substrates are acetate, propanoate, butyrate, 4-hydroxybenzoate, jasmonate, cinnamate, coumarate, and caffeate | Clarkia breweri | ? | - |
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|
S-adenosyl-L-methionine + 3-hydroxybenzoate | 26% activity compared to salicylate | Clarkia breweri | methyl 3-hydroxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + benzoate | 96% activity compared to salicylate | Clarkia breweri | methyl benzoate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + salicylate | - |
Clarkia breweri | methyl salicylate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + salicylate | best substrate | Clarkia breweri | methyl salicylate + S-adenosyl-L-homocysteine | - |
? | |
S-adenosyl-L-methionine + vanillate | 12% activity compared to salicylate | Clarkia breweri | methyl vanillate + S-adenosyl-L-homocysteine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
More | see also EC 2.1.1.273 | Clarkia breweri |
SA MTase | - |
Clarkia breweri |
salicylic acid methyltransferase | - |
Clarkia breweri |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Clarkia breweri |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0163 | - |
salicylate | pH 7.5, 30°C | Clarkia breweri |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Clarkia breweri |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Clarkia breweri |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.4 | - |
salicylate | pH 7.5, 30°C | Clarkia breweri |