PMID: 15388964Sep 25, 2004Paper

Purification and characterization of glutamine synthetase of Pseudomonas taetrolens Y-30: an enzyme usable for production of theanine by coupling with the alcoholic fermentation system of baker's yeast

Bioscience, Biotechnology, and Biochemistry
Sachiko YamamotoT Tachiki

Abstract

Concentrated cell-extract of Pseudomonas taetrolens Y-30, isolated as a methylamine-assimilating organism, formed gamma-glutamylethylamide (theanine) from glutamic acid and ethylamine in a mixture containing the alcoholic fermentation system of baker's yeast for ATP-regeneration. Glutamine synthetase (GS), probably responsible for theanine formation, was isolated from the extract of the organism grown on a medium containing 1% methylamine, 1% glycerol, 0.5% yeast extract, and 0.2% polypepton as carbon and nitrogen sources. The molecular mass was estimated to be 660 kDa by gel filtration and 55 kDa by SDS-polyacrylamide gel electrophoresis, suggesting that Ps. taetrolens Y-30 GS consists of 12 identical subunits. The enzyme required Mg2+ or Mn2+ for its activity. Under the standard reaction condition for glutamine formation (pH 8.0 with 30 mM Mg2+), GS showed 7% and 1% reactivity toward methylamine and ethylamine respectively of that to ammonia. Reactivity to the alkylamines varied with optimum pH of the reaction in response to divalent cation in the mixture: pH 11.0 was the optimum for the Mg2+ -dependent reaction with ethylamine, and pH 8.5 was the optimum for the Mn2+ -dependent reaction. In a mixture of an optimum reaction c...Continue Reading

References

Nov 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·W B AndersonE R Stadtman

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Citations

Feb 8, 2007·Bioscience, Biotechnology, and Biochemistry·Sachiko YamamotoTakashi Tachiki
Dec 8, 2009·Bioscience, Biotechnology, and Biochemistry·Koichiro Miyake, Shingo Kakita
May 4, 2005·Yeast
Nov 30, 2019·Applied Microbiology and Biotechnology·Hongkun MaYanjun Li
Jan 22, 2021·Journal of Agricultural and Food Chemistry·Zhen ChenNing He
Mar 21, 2021·Applied and Environmental Microbiology·Ryota HagiharaHirofumi Endo
Sep 2, 2021·Journal of Agricultural and Food Chemistry·Leonie BenninghausVolker F Wendisch

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