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Literature summary extracted from

  • Yamamoto, S.; Wakayama, M.; Tachiki, T.
    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.
    View publication on PubMed

Application

EC Number Application Comment Organism
6.3.4.12 synthesis the enzyme is usable in theanine formation by coupling with the alcoholic fermentation system of bakers yeast, Saccharomyces cerevisiae Methylovorus mays

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.4.12 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

EC Number Metals/Ions Comment Organism Structure
6.3.4.12 Mg2+
-
Methylovorus mays

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.4.12 49000
-
x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE Methylovorus mays
6.3.4.12 50000
-
x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE Methylovorus mays

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.4.12 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
-
?
6.3.4.12 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
-
?
6.3.4.12 ATP + L-glutamate + methylamine Methylovorus mays N5-methyl-L-glutamine is probably a temporary storage sink for methylamine ADP + phosphate + N5-methyl-L-glutamine
-
?
6.3.4.12 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
-
?
6.3.4.12 additional information Methylovorus mays the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction ?
-
?
6.3.4.12 additional information Methylovorus mays No. 9 the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction ?
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.3.4.12 Methylovorus mays
-
-
-
6.3.4.12 Methylovorus mays No. 9
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.4.12 recombinant enzyme from Escherichia coli strain AD494(DE3) by ammonium sulfate fractionation and anion exchange chromatography Methylovorus mays

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.4.12 8
-
purified recombinant enzyme Methylovorus mays

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.4.12 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
-
?
6.3.4.12 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
-
?
6.3.4.12 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
-
?
6.3.4.12 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
-
?
6.3.4.12 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
-
?
6.3.4.12 ATP + L-glutamate + methylamine N5-methyl-L-glutamine is probably a temporary storage sink for methylamine Methylovorus mays ADP + phosphate + N5-methyl-L-glutamine
-
?
6.3.4.12 ATP + L-glutamate + methylamine preferred substrate of the native and recombinant enzymes Methylovorus mays ADP + phosphate + N5-methyl-L-glutamine
-
?
6.3.4.12 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
-
?
6.3.4.12 additional information the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction Methylovorus mays ?
-
?
6.3.4.12 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 ?
-
?
6.3.4.12 additional information the isolated theanine-forming enzyme predominantly catalyzes the gamma-glutamyl-methylamide forming reaction Methylovorus mays No. 9 ?
-
?
6.3.4.12 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

EC Number Subunits Comment Organism
6.3.4.12 ? x * 49000, about, sequence calculation, x * 50000, recombinnat enzyme, SDS-PAGE Methylovorus mays

Synonyms

EC Number Synonyms Comment Organism
6.3.4.12 gamma-Glutamylmethylamide synthetase
-
Methylovorus mays
6.3.4.12 GMAS
-
Methylovorus mays

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.4.12 40
-
recombinant enzyme Methylovorus mays

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.4.12 7.5 8 recombinant and native enzymes Methylovorus mays

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
6.3.4.12 6
-
the native as well as the recombinant enzyme is most stable at pH 6.0 Methylovorus mays

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.4.12 ATP
-
Methylovorus mays