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Literature summary for 6.3.4.12 extracted from
- Cloning and expression of Methylovorus mays No. 9 gene encoding gamma-glutamylmethylamide synthetase: an enzyme usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast (2008), Biosci. Biotechnol. Biochem., 72, 101-109.
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | the enzyme is usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae | Methylovorus mays |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| cloning from a genomic DNA library, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain AD494(DE3), subcloning in Escherichia coli strain JM109 | Methylovorus mays |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Methylovorus mays |  |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 49000 | - |
x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE | Methylovorus mays |
| 50000 | - |
x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE | Methylovorus mays |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-glutamate + ethylamine | Methylovorus mays | the enzyme is involved in the theanine biosynthesis and usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae. Theanine, gamma-glutamyl-ethylamide, is a nonproteininc amino acid | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + ethylamine | Methylovorus mays No. 9 | the enzyme is involved in the theanine biosynthesis and usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae. Theanine, gamma-glutamyl-ethylamide, is a nonproteininc amino acid | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + methylamine | Methylovorus mays | N5-methyl-L-glutamine is probably a temporary storage sink for methylamine | ADP + phosphate + N5-methyl-L-glutamine | - |
? | |
| ATP + L-glutamate + methylamine | Methylovorus mays No. 9 | N5-methyl-L-glutamine is probably a temporary storage sink for methylamine | ADP + phosphate + N5-methyl-L-glutamine | - |
? | |
| additional information | Methylovorus mays | the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction | ? | - |
? | |
| additional information | Methylovorus mays No. 9 | the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Methylovorus mays | - |
- |
- |
| Methylovorus mays No. 9 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme from Escherichia coli strain AD494(DE3) by ammonium sulfate fractionation and anion exchange chromatography | Methylovorus mays |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 8 | - |
purified recombinant enzyme | Methylovorus mays |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-glutamate + ethylamine | the enzyme is involved in the theanine biosynthesis and usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae. Theanine, gamma-glutamyl-ethylamide, is a nonproteininc amino acid | Methylovorus mays | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + ethylamine | 72-75% activity of the activity with methylamine for the native and recombinant enzymes | Methylovorus mays | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + ethylamine | the enzyme is involved in the theanine biosynthesis and usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae. Theanine, gamma-glutamyl-ethylamide, is a nonproteininc amino acid | Methylovorus mays No. 9 | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + ethylamine | 72-75% activity of the activity with methylamine for the native and recombinant enzymes | Methylovorus mays No. 9 | ADP + phosphate + N5-ethyl-L-glutamine | - |
? | |
| ATP + L-glutamate + hydroxylamine | 30-40% activity of the activity with methylamine for the native and recombinant enzymes | Methylovorus mays | ADP + phosphate + N5-hydroxy-L-glutamine | - |
? | |
| ATP + L-glutamate + methylamine | N5-methyl-L-glutamine is probably a temporary storage sink for methylamine | Methylovorus mays | ADP + phosphate + N5-methyl-L-glutamine | - |
? | |
| ATP + L-glutamate + methylamine | preferred substrate of the native and recombinant enzymes | Methylovorus mays | ADP + phosphate + N5-methyl-L-glutamine | - |
? | |
| ATP + L-glutamate + methylamine | N5-methyl-L-glutamine is probably a temporary storage sink for methylamine | Methylovorus mays No. 9 | ADP + phosphate + N5-methyl-L-glutamine | - |
? | |
| additional information | the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction | Methylovorus mays | ? | - |
? | |
| additional information | ammonia is a poor substrate showing 0.5-0.75% of the activity with methylamine for the native and recombinant enzymes | Methylovorus mays | ? | - |
? | |
| additional information | the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction | Methylovorus mays No. 9 | ? | - |
? | |
| additional information | ammonia is a poor substrate showing 0.5-0.75% of the activity with methylamine for the native and recombinant enzymes | Methylovorus mays No. 9 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE | Methylovorus mays |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| gamma-Glutamylmethylamide synthetase | - |
Methylovorus mays |
| GMAS | - |
Methylovorus mays |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
recombinant enzyme | Methylovorus mays |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 8 | recombinant and native enzymes | Methylovorus mays |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
the native as well as the recombinant enzyme is most stable at pH 6.0 | Methylovorus mays |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Methylovorus mays | |
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