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Literature summary for 2.1.1.B126 extracted from
- Benzenoid biosynthesis in the flowers of Eriobotrya japonica molecular cloning and functional characterization of p-methoxybenzoic acid carboxyl methyltransferase (2016), Planta, 244, 725-736 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene EjMT1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic tree, quantitative RT-PCR enzyme expression analysis, functional recombinant expression of His-tagged enzyme EjMT1 in Escherichia coli | Rhaphiolepis bibas |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0091 | - |
S-adenosyl-L-methionine | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas |  |
| 0.1373 | - |
4-Methoxybenzoate | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| S-adenosyl-L-methionine + 4-methoxybenzoate | Rhaphiolepis bibas | - |
methyl 4-methoxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rhaphiolepis bibas | A0A1B4Z3V0 | flowers and leaves collected in winter at the Uji campus (E135°48', N34°54') of Kyoto University, Japan | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme EjMT1 from Escherichia coli by nickel affinity chromatography | Rhaphiolepis bibas |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| flower | inflorescences of loquat flowers constitute stiff panicles, open flowers are classified as one of four developmental stages, stage 1, stage 2, stage 3, or stage 4. Transcript levels of MBMT continually increase throughout the flower development with peak expression occurring in fully opened flowers | Rhaphiolepis bibas | - |
| leaf | low enzyme level | Rhaphiolepis bibas | - |
| additional information | the floral trichomes do not contribute to the production of volatile benzenoids in loquat. EjMT1 is highly expressed in flower petals as compared to the other three EjMT genes. The transcript levels of EjMT1 gene increase as buds matured into flowers and those of EjMT1 are 100 to 300folds higher in flower tissues than in leaves. Quantitative enzyme expression analysis by RT-PCR | Rhaphiolepis bibas | - |
| petal | - |
Rhaphiolepis bibas | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | recombinant MBMT protein expressed in Escherichia coli shows the highest substrate preference toward 4-methoxybenzoic acid. EjMT1 shows high activity with 3-methoxybenzoic acid and vanillic acid. The activity of EjMBMT with benzoic acid, 2-methoxybenzoic acid, anthranilic acid, and jasmonic acid is approximately half of that with p-methoxybenzoic acid. Enzyme EjMBMT displays negligible activity to salicylic acid, shikimic acid, and caffeic acid, no activity with indole-3-acetic acid and cinnamic acid. The enzyme is relatively specific for methoxybenzoic acids and the methoxy group on a benzene ring might be important for either enzyme binding or methyl group transfer from SAM by EjMBMT | Rhaphiolepis bibas | ? | - |
- |
|
| S-adenosyl-L-methionine + 2-methoxybenzoate | low activity | Rhaphiolepis bibas | methyl 2-methoxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + 3-methoxybenzoate | high activity | Rhaphiolepis bibas | methyl 3-methoxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + 4-methoxybenzoate | - |
Rhaphiolepis bibas | methyl 4-methoxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + 4-methoxybenzoate | best substrate | Rhaphiolepis bibas | methyl 4-methoxybenzoate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + anthranilate | low activity | Rhaphiolepis bibas | methyl anthranilate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + jasmonate | low activity | Rhaphiolepis bibas | methyl jasmonate + S-adenosyl-L-homocysteine | - |
? | |
| S-adenosyl-L-methionine + vanillate | - |
Rhaphiolepis bibas | methyl vanillate + S-adenosyl-L-homocysteine | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| EjMBMT | - |
Rhaphiolepis bibas |
| EjMT1 | - |
Rhaphiolepis bibas |
| MBMT | - |
Rhaphiolepis bibas |
| p-methoxybenzoic acid carboxyl methyltransferase | - |
Rhaphiolepis bibas |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Rhaphiolepis bibas |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.018 | - |
S-adenosyl-L-methionine | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas | |
| 0.018 | - |
4-Methoxybenzoate | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 6.5 | recombinant enzyme | Rhaphiolepis bibas |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 7.5 | activity range, recombinant enzyme | Rhaphiolepis bibas |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| S-adenosyl-L-methionine | - |
Rhaphiolepis bibas | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | in contrast to benzoic acid carboxyl methyltransferase (BAMT) and benzoic acid/salicylic acid carboxyl methyltransferase (EC 2.1.1.274/2.1.1.273), MBMT also displays activity towards both benzoic acid and jasmonic acid. Phylogenetic analysis reveals that loquat MBMT forms a monophyletic group with jasmonic acid carboxyl methyltransferases (JMTs, EC 2.1.1.141) from other plant species. Plant enzymes with same BAMT activity have evolved independently | Rhaphiolepis bibas |
| metabolism | it appears likely that a para-hydroxyl group of p-hydroxybenzoic acid is first methylated by an yet unidentified O-methyltransferase, then EjMBMT converts the resulting p-methoxybenzoic acid to methyl 4-methoxybenzoate in vivo, but not vice versa | Rhaphiolepis bibas |
| physiological function | EjMT1 encodes a p-methoxybenzoic acid carboxyl methyltransferase (MBMT), an enzyme capable of converting p-methoxybenzoic acid to methyl p-methoxybenzoate via methylation of the carboxyl group. 4-Methoxybenzaldehyde and methyl 4-methoxybenzoate, along with (2-nitroethyl) benzene, are major components of the floral scent of the Japanese loquat (Eriobotrya japonica). Distribution of volatile benzenoids in different parts of the loquat plant, overview | Rhaphiolepis bibas |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.131 | - |
4-Methoxybenzoate | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas | |
| 1.98 | - |
S-adenosyl-L-methionine | pH 6.5, 25°C, recombinant enzyme | Rhaphiolepis bibas | |
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