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Yuan Li, Shan Liu, Jun Zhu
Recombinant strains expressing enzymes for ATP regeneration and L-theanine production were constructed and used for the synthesis of L-theanine. The ppk gene encoding polyphosphate kinase (PPK) from Rhodobacter sphaeroides and gmas gene encoding γ-glutamylmethylamide synthetase (GMAS) from Methylovorus mays were synthesized, and two recombinant plasmids, pETDuet-ppk+gmas and pET21a-ppk+gmas were constructed for co-expression of PPK and GMAS in Escherichia coli BL21(DE3). SDS-PAGE analysis showed that PPK and GMAS were overexpressed in soluble form in both recombinant strains...
December 25, 2016: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Wanmeng Mu, Tao Zhang, Bo Jiang
L-Theanine (γ-glutamylethylamide) is a unique non-protein amino acid that is naturally found in tea plants. It contributes to the umami taste and unique flavor to green tea infusion, and thus its content in tea leaves highly impacts the tea quality and price. In addition to the graceful taste, it has been proved to have many beneficial physiological effects, especially promoting relaxation and improving concentration and learning ability. Based on these promising advantages, L-theanine has been commercially developed as a valuable ingredient for use in food and beverages to improve and/or maintain human health...
May 2015: Biotechnology Advances
Dipti D Nayak, Christopher J Marx
Methylotrophs grow on reduced single-carbon compounds like methylamine as the sole source of carbon and energy. In Methylobacterium extorquens AM1, the best-studied aerobic methylotroph, a periplasmic methylamine dehydrogenase that catalyzes the primary oxidation of methylamine to formaldehyde has been examined in great detail. However, recent metagenomic data from natural ecosystems are revealing the abundance and importance of lesser-known routes, such as the N-methylglutamate pathway, for methylamine oxidation...
December 2014: Journal of Bacteriology
Daniela Wischer, Deepak Kumaresan, Antonia Johnston, Myriam El Khawand, Jason Stephenson, Alexandra M Hillebrand-Voiculescu, Yin Chen, J Colin Murrell
Movile Cave, Romania, is an unusual underground ecosystem that has been sealed off from the outside world for several million years and is sustained by non-phototrophic carbon fixation. Methane and sulfur-oxidising bacteria are the main primary producers, supporting a complex food web that includes bacteria, fungi and cave-adapted invertebrates. A range of methylotrophic bacteria in Movile Cave grow on one-carbon compounds including methylated amines, which are produced via decomposition of organic-rich microbial mats...
January 2015: ISME Journal
Lisheng Xu, Guizhen Gao, Cao Wengen, Jigui Xu, Liang Zhao, Hongwei Shi, Xingtao Zhang
A new method for the synthesis of γ-glutamylmethylamide is presented. Glutamic acid γ-methyl ester was used as substrate for γ-glutamylmethylamide synthesis catalyzed by Escherichia coli with γ-glutamyltranspeptidase activity. Reaction conditions were optimized by using 300 mM glutamic acid γ-methyl ester and 3,000 mM methylamine at pH 10 and 40 °C. Bioconversion rate of γ-glutamylmethylamide reached 87 % after 10 h. γ-Glutamyltranspeptidase was reversibly inhibited only when glutamic acid γ-methyl ester was above 300 mM...
June 2014: Applied Biochemistry and Biotechnology
Christelle Gruffaz, Emilie E L Muller, Yousra Louhichi-Jelail, Yella R Nelli, Gilles Guichard, Françoise Bringel
Monomethylamine (MMA, CH3NH2) can be used as a carbon and nitrogen source by many methylotrophic bacteria. Methylobacterium extorquens DM4 lacks the MMA dehydrogenase encoded by mau genes, which in M. extorquens AM1 is essential for growth on MMA. Identification and characterization of minitransposon mutants with an MMA-dependent phenotype showed that strain DM4 grows with MMA as the sole source of carbon, energy, and nitrogen by the N-methylglutamate (NMG) pathway. Independent mutations were found in a chromosomal region containing the genes gmaS, mgsABC, and mgdABCD for the three enzymes of the pathway, γ-glutamylmethylamide (GMA) synthetase, NMG synthase, and NMG dehydrogenase, respectively...
June 2014: Applied and Environmental Microbiology
Nina V Doronina, Elena N Kaparullina, Tatjana V Bykova, Yuri A Trotsenko
A newly isolated, facultatively methylotrophic bacterium (strain MUSA(T)) was investigated. The isolate was strictly aerobic, Gram-stain-negative, asporogenous, motile, rod-shaped and multiplied by binary fission. The strain utilized methanol, methylamine and an apparently narrow range of multi-carbon compounds, but not methane, dichloromethane or CO2/H2, as the carbon and energy sources. Growth occurred at pH 5.5-9.5 (optimum, pH 7.0) and 16-40 °C (optimum, 28-30 °C). The major fatty acids of methanol-grown cells were C18 : 1ω7c, C18 : 0 and 11-methyl-C18 : 1ω7c ...
May 2013: International Journal of Systematic and Evolutionary Microbiology
Yin Chen
The marine Roseobacter clade bacteria comprise up to 20% of the microbial community in coastal surface seawater. Marine Roseobacter clade bacteria are known to catalyse some important biogeochemical transformations in marine carbon and sulfur cycles. Using a comparative genomic approach, this study revealed that many marine Roseobacter clade bacteria have the genetic potential to utilize methylated amines (MAs) as alternative nitrogen sources. These MAs represent a significant pool of dissolved organic carbon and nitrogen in the marine environment...
September 2012: Environmental Microbiology
Yin Chen, Julie Scanlan, Lijiang Song, Andrew Crombie, M Tanvir Rahman, Hendrik Schäfer, J Colin Murrell
Methylocella silvestris BL2, a facultative methane utilizer, can grow on monomethylamine (MMA) as a sole carbon and nitrogen source. No activity of MMA dehydrogenase was detectable. Instead, this bacterium utilizes a methylated amino acid pathway (gamma-glutamylmethylamide [GMA] and N-methylglutamate [NMG]) for MMA metabolism. The activities of the two key enzymes in this pathway, GMA synthetase and NMG dehydrogenase, were found when the bacterium was grown on MMA. GMA was detected by high-performance liquid chromatography-mass spectrometry only when the bacterium was grown on MMA but not when it was grown on methanol...
July 2010: Applied and Environmental Microbiology
Yin Chen, Kathryn L McAleer, J Colin Murrell
Monomethylamine can be used by nonmethylotrophs as a sole nitrogen source but not as a carbon source; however, little is known about the genes and enzymes involved. The gamma-glutamylmethylamide/N-methylglutamate pathway for monomethylamine utilization by methylotrophs has recently been resolved. We have identified genes encoding key enzymes of this pathway in nonmethylotrophs (e.g., Agrobacterium tumefaciens) and demonstrated that this pathway is also involved in the utilization of monomethylamine as a nitrogen source by nonmethylotrophs...
June 2010: Applied and Environmental Microbiology
Sachiko Yamamoto, Yosuke Morihara, Mamoru Wakayama, Takashi Tachiki
Gamma-glutamylmethylamide synthetase and dried baker's yeast cells were enclosed together in a dialysis membrane tube to produce theanine repeatedly by coupled fermentation with energy transfer. The membrane-enclosed enzyme preparation (M-EEP) formed approximately 600 mM theanine from glutamic acid and ethylamine at a 100% conversion rate. M-EEP maintained its productivity of theanine during six consecutive reactions in a mixture containing NAD(+).
December 2009: Bioscience, Biotechnology, and Biochemistry
Ekaterina Latypova, Song Yang, Yi-Shun Wang, Tiansong Wang, Theodore A Chavkin, Murray Hackett, Hendrik Schäfer, Marina G Kalyuzhnaya
The ability of some microbial species to oxidize monomethylamine via glutamate-mediated pathways was proposed in the 1960s; however, genetic determinants of the pathways have never been described. In the present study we describe a gene cluster essential for operation of the N-methylglutamate pathway in the methylotrophic beta-proteobacterium Methyloversatilis universalis FAM5. Four major polypeptides from protein fractions displaying high activities of N-methylglutamate synthetase, N-methylglutamate dehydrogenase and gamma-glutamylmethylamide synthetase were selected for mass spectrometry-based identification...
January 2010: Molecular Microbiology
Sachiko Yamamoto, Yosuke Morihara, Mamoru Wakayama, Takashi Tachiki
Gamma-glutamylmetylamide synthetase (GMAS) of Methylovorus mays No. 9, produced by Eschericia coli AD494 (DE3) harboring pET21aGM, formed theanine from glutamic acid and ethylamine with coupling of the reaction with sugar fermentation of baker's yeast cells as an ATP-regeneration system. Theanine formation was stimulated by the addition of Mn(2+) to the reaction mixture, whereas Mg(2+) was less effective. Increases to a certain level in the concentrations of GMAS and the substrates in the mixture were effective in increasing theanine formation, but high concentrations of ethylamine (900 mM or more) inhibited yeast sugar fermentation, and eventually decreased theanine formation...
May 2008: Bioscience, Biotechnology, and Biochemistry
Sachiko Yamamoto, Mamoru Wakayama, Takashi Tachiki
Gamma-glutamylmethylamide synthetase (GMAS), found in an obligate methylotroph, Methylovorus mays No. 9, can form theanine from glutamic acid and ethylamine in a mixture in which yeast sugar fermentation is coupled for ATP regeneration. The internal and N-terminal amino acid sequences of GMAS had certain similarities to putative glutamine synthetase type III (GS III) of Methylobacillus flagellatus KT. From the M. mays No. 9 genomic DNA library, a clone containing a 6.5-kbp insertional DNA fragment was selected by the PCR screening technique with oligonucleotide primers specific for the GMAS gene...
January 2008: Bioscience, Biotechnology, and Biochemistry
Sachiko Yamamoto, Mamoru Wakayama, Takashi Tachiki
For development of theanine production from glutamic acid and ethylamine by coupling yeast sugar fermentation as an ATP-regenerating system, several strains were selected from among about 200 methylamine- and/or methanol-assimilating bacteria depending on the theanine-forming activity of their permeated cells. The amount of theanine formed by the cells of the selected strains was much larger than that by the cells of Escherichia coli AD494 (DE3) expressing Pseudomonas taetrolens Y-30 glutamine synthetase (GS), which has been found to be a usable enzyme for theanine production...
February 2007: Bioscience, Biotechnology, and Biochemistry
No abstract text is available yet for this article.
February 1954: Archives of Biochemistry and Biophysics
A N Galkin, L E Mikheeva, S V Shestakov
Synechocystis sp. PCC 6803 mutants, in which one of the eukaryotic-type serine/threonine protein kinase genes pknD, pknE, pknG, and pknH was inactivated, were obtained by insertion mutagenesis. None of these mutants differed phenotypically from the wild-type strain, indicating that the pknD, pknE, pknG, and pknH genes are not of crucial importance for the photoautotrophically grown cyanobacterium. Mutant with the inactivated pknE gene was resistant to L-methionine-D,L-sulfoximine and especially to methylamine...
January 2003: Mikrobiologiia
N V Doronina, Y A Trotsenko, T P Tourova
A new genus, Methylarcula, with two new species, Methylarcula marina and Methylarcula terricola, are proposed for strains h1T and h37T of moderately halophilic facultatively methylotrophic bacteria isolated from the coastal saline habitats. These methylobacteria are aerobic, Gram-negative, asporogenous, non-motile, colourless rods that multiply by binary fission. Their cellular fatty acids profiles consist primarily of straight-chain unsaturated (C18:1; 70-80%), saturated (C18:0; 14-16%) and cyclopropane (C19:0; 5-6%) acids...
September 2000: International Journal of Systematic and Evolutionary Microbiology
H Yokogoshi, M Kobayashi
The effect of gamma-glutamylmethylamide(GMA), one of the components of green tea extract, on the blood pressure in spontaneously hypertensive rats (SHR) was investigated. The effect of glutamic acid and r-glutamylethylamide (theanine), which is structurally similar to GMA, was also examined. When SHR were injected with glutamic acid (2000mg/kg), the blood pressure was not altered. The same dose of theanine decreased it significantly. GMA administration to SHR reduced the blood pressure significantly, and its degree of hypotensive action was more effective than that by theanine administration...
1998: Life Sciences
A Jayakumar, E M Barnes
Under N2-fixing conditions, Azotobacter vinelandii expresses a specific transport system for methylammonium (ammonium) [E. M. Barnes, Jr. and P. Zimniak (1981) J. Bacteriol. 146, 512-516]. This activity is decreased markedly by culture of cells in the presence of 10 mM ammonium or 2 mM methylammonium; in both cases, the Vmax values for methylammonium uptake were 25% of those of N2-fixing cells. Mixing experiments with assay medium indicate that transport activity is controlled by intracellular rather than extracellular metabolites...
May 15, 1984: Archives of Biochemistry and Biophysics
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