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Activities and kinetic characteristics of glutamine synthetase fromPenicillium cyclopium

  • Industrial Microbiology
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Abstract

The level of glutamine synthetase (GS; EC 6.3.1.2) obtained as a function of culture age ofPenicillium cyclopium showed that GS activity increased continuously up to 120 hours after which the enzyme activity started to decline. Glutamine synthetase fromP. cyclopium was purified to homogeneity by ammonium sulphate, diethylaminoethyl (DEAE)-cellulose and Sepharose 4B. The purified enzyme showed a single band. The pH optima for both the biosynthetic and transferase activities of the enzyme were 8.2 and 8.5, respectively. Various products such as 5′-AMP, tryptophan, alanine, glycine, and histidine inhibited both reactions of GS. However, ADP activated the transferase, reaction and inhibited the biosynthetic one. The Km values for L-glutamine were 1.4 and 5.6 mM in the biosynthetic and transferase assays, respectively. The divalent metal ion is necessary for the activity of the enzyme. Mn2+ was the most effective metal ion for transferase activity however, Mg2+ was the most effective metal ion for biosynthetic activity. Ca2+ and Mn2+ strongly inhibited Mg2+-supported biosynthetic activity. GS was quite stable in tris (hydroxymethyl) aminomethane-HCl buffer (pH 7.5) containing ethylenediaminetetraacetate (EDTA), MgCl2 and 2-mercaptoethanol (2-ME).

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References

  • Bespalova L.A., Antonyuk L.P., Ignatov L. (1999).Azospirillum brasilense glutamine synthetase: influence of the activating metal ions on the enzyme properties. BioMetal, 12: 115–121.

    Article  CAS  Google Scholar 

  • Bhatnagar L., Zeikus J.G., Aubert J.P. (1986). Purification and characterization of glutamine synthetase from theArchaebacterium ethanobavterium. J. Bacteriol., 165: 638–643.

    CAS  PubMed  Google Scholar 

  • Blanco F., Alalia A., Llama M.J., Serra J.L. (1989). Purification and properties of glutamine synthetase from the non-N2-fixing cyanobacteriumPhormidium laminosum. J. Bacteriol., 171: 1158–1165.

    CAS  PubMed  Google Scholar 

  • Bradford M.M. (1976). A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72: 248–257.

    Article  CAS  PubMed  Google Scholar 

  • Brun A., Chalot M., Botton B., Martin F. (1992). Purification and characterization of NADP-gluatamate dehydrogenase from the ectomycorrhizal fungusLaccaria laccata. Plant Physiol., 99: 938–944.

    Article  CAS  PubMed  Google Scholar 

  • Brun A., Chalot M., Duponnois R., Botton B., Dexheimer J. (1995). Immunogold localization of glutamine synthetase and NADP-glutamate dehydrogenase ofLaccaria laccata in Douglas fir ectomycorrhizas. Mycorrhiza, 5: 139–144.

    Article  Google Scholar 

  • Carvalho H., Lescure N., Billy F., Chabaud M., Lima L., Salema R., Cullimore J., Billy F. (2000). Cellular expression and regulation of theMedicago truncatula cytosolic glutamine synthetase genes in root nodules. Plant Molec. Biol., 42: 471–756.

    Google Scholar 

  • Chen J., Kennedy I.R. (1985). Purification and properties of lupin nodule glutamine synthetase. Phytochemistry, 24: 2167–2172.

    Article  CAS  Google Scholar 

  • Clemente M.T., Marquez A.L. (1999).Phaseolus vulgaris glutamine synthetase alters kinetics and structural properties and confers resistance to L-methionine sulfoximine. Plant Mol. Biol., 40: 835–854.

    Article  CAS  PubMed  Google Scholar 

  • El-Shora H.M., El-Naggar M.E., Shaaban S.A., Zaid A.M. (1994a). Studies on regulatory functions of glutamine synthetase of differentSargassum species. J. Agric. Sci. Fac. Agric. Mansoura Univ., 19 (12): 4311–4326.

    Google Scholar 

  • El-Shora H.M., El-Naggar M.E., Shaaban S.A., Zaid A.M. (1994b). Kinetic properties of glutamine synthetase of differentSargassum species. J. Environ. Sci., 10: 173–180.

    Google Scholar 

  • El-Shora H.M. (1995). Presence of histidine in glutamine synthetase from the roots of carrot, turnip and radish J. Environ. Sci., 10: 159–172.

    CAS  Google Scholar 

  • El-Shora H.M. (2001). Effect of growth regulators and group-modifiers on NADH-glutamate synthase of marrow cotyledons. J. Biol. Sci., 7: 597–602.

    Google Scholar 

  • Florencio F.J., Ramos J.L. (1985). Purification and characterization of glutamine synthetase from th unicellula cyanobacteriumAnacysti nidulans. Biochem. Biophys. Acta, 838: 39–48.

    CAS  Google Scholar 

  • Guerrero M.O., Lara C. (1987). Assimilation of inorganic nitrogen. In: Fay P., Van-Baslen C., Eds, The Cyanobacteria, Elsevier, Amsterdam, pp. 163–186.

    Google Scholar 

  • Ip M.S., Rowell P., Stewart W.D.P. (1983). The role of specific cations in regulation of cyanobacterial glutamine synthetase. Biochem. Biophys. Res. Com., 114: 206–213.

    Article  CAS  PubMed  Google Scholar 

  • Jukim Y., Yoshizawa M., Takemala S., Murakami S., Aoisi K. (2002). Ammonia asimilation inKlebsiella pneumoniae F-5-2 that can utilize ammonium and nitrate ions simultaneously: Purification and characterization of glutamate dehydrogenase and glutamine synthetase. J. Biosci. Bioengin., 93 (6): 584–588.

    Google Scholar 

  • Kameya F., Arai H., Ishii M., Igarashi Y. (2006). Purification and properties of glutamine synthetase fromHydrogenobacter thermophilus TK-6. J. Biosci. Bioengin., 102: (4): 311–315.

    Article  CAS  Google Scholar 

  • Kersten M., Drift C.V., Camp H. D., Baars J.P., Griensven L.V., Schaap P.J., Muller Y., Visser J.A. (2000). The glutamine synthetase from the edible mushroomAgaricus bisporus. International Congress on The Science and Cultivation of Edible Fungi, Maastricht, Netherlands, 15–19 May, 45: 71–78.

  • Laemmli U.K. (1970). Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature, 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  • Liorca O., Betti M., Gonzalez J.M., Valencia A., Marquez A.J. (2006). The three-dimensional structure of an eukaryotic glutamine synthetase: Functional implications of its oligomeric structure. J. Struct. Biol., 156: 469–479.

    Article  Google Scholar 

  • Mash C., Muller Y., Camp H.O.D., Volgels G.D., Griensven L.V., Visser J., Schaap P.J., Van-Griensven L.D. (1997). Molecular characterization of the gInA gene encoding glutamine synthetase from the edible mushroomAgaricus bisporus. Mol. Gen. Genet., 256: 179–186.

    Google Scholar 

  • Mash K., Van-der D., Den C.H., Baars J.J.P., Van G.L., Schaap P.J., Muller Y., Visser J., Jam L., Van, G. (2000). The glutamine synthetase from edible mushroomAgaricus bisporus. International Congress on The Science and Cultivation of Edible Fungi, Mash, Netherlands, 15–19 May, pp. 71–78.

  • MeMaster B.J., Danton M.S., Storch T.A., Dunham V.L. (1980). Regulation of glutamine synthetase in the blue green algaAnabaena flosaquae. Biochem. Biophys. Res. Com., 96: 975–983.

    Article  Google Scholar 

  • Merida A., Leurentop L., Candau P., Florencio F.J. (1990). Purification and properties of glutamine synthetase from the cyanobacteriaSynechocystis sp. strain PCC 6803 andCalothrix sp. strain PCC 7601. J. Bacteriol., 172: 4732–4735.

    CAS  PubMed  Google Scholar 

  • Oliveria I.C., Coruzzi G.M. (1999). Carbon and amino acids reciprocally modulate the expression of glutamine synthetase inArabidopsis. Plant Physiol., 121: 301–309.

    Article  Google Scholar 

  • Orr J., Haselkorn R. (1981). Kinetic and inhibition studies of glutamine synthetase from the cyanobacteriumAnabaena 7120. J. Biol. Chem., 256: 13099–13104.

    CAS  PubMed  Google Scholar 

  • Orr L.M., Keefer P., Keim T.D., Nguyen T., Wellems R.L., Haselkorn R. (1981). Purification, physical characterization and NH2-terminal sequence of glutamine synthetase from the cyanobacteriumAnabaena 7120.3. Biol. Chem., 256: 13091–13098.

    CAS  Google Scholar 

  • Ortega J.L., Roche D., Sengupta G.C. (1999). Oxidative turnover of soybean root glutamine synthetase.In vitro andin vivo studies. Plant Physiol., 119: 1483–1495.

    Article  CAS  PubMed  Google Scholar 

  • Palaniappan C., Gunasekaran M. (1995). Purification and properties of glutamine synthetase fromNocardia asteroides. Curr. Microbiol., 31: 193–198.

    Article  CAS  PubMed  Google Scholar 

  • Palmer T. (1985). Understanding enzymes. Ellis Horwood Publishers, England.

    Google Scholar 

  • Rudolfova V., Mikes V. (1997). Involvement of the glutamine synthetase/glutamate synthase pathway in ammonia assimilation by the wood-rotting fungusPleurotus ostreatus. Folia Microbiol., 42: 577–582.

    Article  CAS  Google Scholar 

  • Segal A., Stadtman E.R. (1972). Variations of the conformational states ofEscherichia coli glutamine synthetase by interaction with different divalent cations. Arch. Biochem. Biophys., 152: 367–377.

    Article  CAS  PubMed  Google Scholar 

  • Shapiro B.M., Stadtman E.R. (1970). The regulation of glutamine synthetase in microorganisms. Annu. Rev. Microbiol., 24: 501–524.

    Article  CAS  PubMed  Google Scholar 

  • Shatters R.G., Liu Y., Khan M.L. (1993). Isolation and characterization of a novel glutamine synthetase fromRhizobium meliloti. J. Biol. Chem., 268: 469–475.

    CAS  PubMed  Google Scholar 

  • Spinosa M., Riccio A., Mandrich L., Manco G., Lamber A., Iaccarino M., Merrick M., Patiarca E.J. (2000). Inhibition of glutamate synthase 11 expression by the product of the gstI gene. Mol. Microbiol., 37: 443–452.

    Article  CAS  PubMed  Google Scholar 

  • Stacey O., Van-Baslen C., Tabita E.R. (1979). Nitrogen and ammonia assimilation in the cyanobacteria: Regulation of glutamine synthetase. Arch. Biochem. Biophys., 194: 457–467.

    Article  CAS  PubMed  Google Scholar 

  • Yadav A.S., Vasudeva M., Upadhyay K.K., Sawhney S.K., Vashishat R.K. (1999). Symbiotic effectiveness, rate of respiration and glutamine synthetase activity of sodium azide-resistant strains ofRhizobium leguminosarum biovartrifolii. Let. Appl. Microbiol., 28: 466–470.

    Article  CAS  Google Scholar 

  • Yuang H.F., Wang C.M., Kung H.W. (2001). Purification and characterization of glutamine synthetase from the unicellular cyanobacteriumSynechococcus RF-1. Bot. Bull. Acad. Sin., 42: 23–33.

    Google Scholar 

  • Zofall M., Schanel L., Turanek J., Camp H. D., Mikes V., Camp O.D. (1996). Purification and characterization of glutamine synthetase from the basidiomycetePleurotus ostreatus. Curr. Microbiol., 33: 181–186.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Botany Department, Faculty of Science, Mansoura University, Mansoura, Egypt

    Hamed Mohammed El-Shora

  2. Botany Department, Faculty of Science, Zagazig University, Zagazig, Egypt

    Salwa AbdelMageed Khalaf

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  1. Hamed Mohammed El-Shora
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  2. Salwa AbdelMageed Khalaf
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Correspondence to Hamed Mohammed El-Shora.

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El-Shora, H.M., Khalaf, S.A. Activities and kinetic characteristics of glutamine synthetase fromPenicillium cyclopium . Ann. Microbiol. 58, 691–696 (2008). https://doi.org/10.1007/BF03175576

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