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Phosphorylation of glucose by a guanosine-5′-triphosphate (GTP)-dependent glucokinase in Fibrobacter succinogenes subsp. succinogenes S85

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Abstract

Cell extracts of Fibrobacter succinogenes subsp. succinogenes S85 phosphorylated glucose with a GTP-dependent glucokinase. The enzyme showed little activity with ATP (12% of that with GTP). Of other phosphate donors tested, only dGTP and ITP gave high glucokinase activities. Dialyzed extracts required Mg+2 and K+ for maximal activity. In potassium phosphate buffer, glucokinase showed maximum activity at pH 7.5 with glucose-6-phosphate dehydrogenase as the coupling enzyme. In this assay, glucokinase was active with glucose (100%), 2-deoxy-d-glucose (40%), and mannose (20%). Partially purified glucokinase had a molecular weight of 82,000 and a pl of 4.82. Double-reciprocal plots of substrate concentration versus velocity were linear and the enzyme had apparent Km values of 55 μM for glucose and 72 μM for GTP. Dialyzed cell extracts of Fibrobacter intestinalis C1A also contained a GTP-dependent glucokinase that showed little activity with ATP. Potassium also stimulated the activity of this enzyme. These results suggest that this unusual glucokinase may be characteristic of the genus Fibrobacter.

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Abbreviations

CHES :

cyclohexylaminoethanesulfonic acid

GK :

glucokinase

PEP :

phosphoenolpyruvate

References

  • Amann RI, Lin C, Key R, Montgomery L, Stahl DA (1992) Diversity among Fibrobacter isolates: towards a phylogenetic classification. System Appl Microbiol 15:23–31

    Google Scholar 

  • Bochner BR, Ames BN (1982) Complete analysis of cellular nucleotides by two dimensional thin layer chromatography. J Biol Chem 257:9759–9769

    Google Scholar 

  • Bryant MP (1972) Commentary on the Hungate technique for culture of anaerobic bacteria. Am J Clin Nutr 25:1321–1328

    Google Scholar 

  • Chow JM, Russell JB (1992) Effect of pH and monensin on glucose transport by Fibrobacter succinogenes, a cellulolytic ruminal bacterium. Appl Environ Microbiol 58:1115–1120

    Google Scholar 

  • Coe EL, Hsu L-H (1973) Acyl coenzyme A inhibition of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase: a comparison of the TPN and DPN linked reactions. Biochem Biophys Res Commun 53:66–69

    Google Scholar 

  • Delvalle JA, Asensio C (1978) Distribution of adenosine 5′-triphosphate (ATP)-dependent hexose kinases in microorganisms. Biosystems 10:265–282

    Google Scholar 

  • Doelle HW (1982a) Kinetic characteristics and regulatory mechanisms of glucokinase and fructokinase from Zymomonas mobilis. Eur J Appl Microbiol Biotechnol 14:241–246

    Google Scholar 

  • Doelle HW (1982b) The existence of two separate constitutive enzymes for glucose and fructose in Zymomonas mobilis. Eur J Appl Microbiol Biotechnol 15:20–24

    Google Scholar 

  • Franklund CV, Glass TL (1987) Glucose uptake by the cellulolytic ruminal anaerobe Bacteroides succinogenes. J Bacteriol 169:500–506

    Google Scholar 

  • Fukuda Y, Yamaguchi S, Shimosaka M, Murata K, Kimura A (1984) Purification and characterization of glucokinase in Escherichia coli B. Agric Biol Chem 48:2541–2548

    Google Scholar 

  • Groleau D, Forsberg CW (1981) Cellulolytic activity of the rumen bacterium Bacteroides succinogenes. Can J Microbiol 27:517–530

    Google Scholar 

  • Hengartner H, Zuber H (1973) Isolation and characterization of a thermophilic glucokinase from Bacillus stearothermophilus. FEBS Lett 37:212–216

    Google Scholar 

  • Horne RN, Anderson WB, Nordlie RC (1970) Glucose dehydrogenase activity of yeast glucose 6-phosphate dehydrogenase. Inhibition by adenosine 5′-triphosphate and other nucleoside 5′-triphosphates and diphosphates. Biochemistry 9:610–616

    Google Scholar 

  • Hungate RE (1969) A roll tube method for cultivation of strict anaerobes. In: Norris JR, Ribbons DW (eds) Methods in microbiology, vol 3. Academic Press, New York, pp 117–132

    Google Scholar 

  • Kalb VF, Bernlohr RW (1977) A new spectrophotometric assay for protein in cell extracts. Anal Biochem 82:362–371

    Google Scholar 

  • Kamel MY, Allison DP, Anderson RL (1966) Stereospecific d-glucokinase of Aerobacter aerogenes. J Biol Chem 241:690–694

    Google Scholar 

  • Maas LK, Glass TL (1991) Cellobiose uptake by the cellulolytic ruminal anaerobe Fibrobacter (Bacteroides) succinogenes. Can J Microbiol 37:141–147

    Google Scholar 

  • Martin SA, Russell JB (1986) Phosphoenolpyruvate-dependent phosphorylation of hexoses by ruminal bacteria: evidence for the phosphotransferase transport system. Appl Environ Microbiol 52:1348–1352

    Google Scholar 

  • Miller TL (1978) The pathway of formation of acetate and succinate from pyruvate by Bacteroides succinogenes. Arch Microbiol 117:145–152

    Google Scholar 

  • Miller TL, Wolin MJ (1974) A serum bottle modification of the Hungate technique for cultivating obligate anaerobes. Appl Microbiol 27:985–987

    Google Scholar 

  • Patni NJ, Alexander JK (1971) Utilization of glucose by Clostridium thermocellum: presence of glucokinase and other glycolytic enzymes in cell extracts. J Bacteriol 105:220–225

    Google Scholar 

  • Porter EV, Casey BM, Holmlund CE (1982) Purification and kinetic characterization of a specific glucokinase from Streptococcus mutans OMZ7O cells. Biochim Biophys Acta 709:178–186

    Google Scholar 

  • Reeves RE, Montaloo F, Sillero A (1967) Glucokinase from Entamoeba histolytica and related organisms. Biochemistry 6:1752–1760

    Google Scholar 

  • Sabater B, Sebastian J, Asenio C (1972) Identification and properties of an inducible mannokinase from Streptomyces violaceoruber. Biochim Biophys Acta 284:406–413

    Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Google Scholar 

  • Stewart CS, Flint HJ (1989) Bacteroides (Fibrobacter) succinogenes, a cellulolytic anaerobic bacterium from the gastrointestinal tract. Appl Microbiol Biotechnol 30:433–439

    Google Scholar 

  • Suelter CH (1970) Enzymes activated by monovalent cations. Science 168:789–795

    Google Scholar 

  • Varel VH, Fryda SJ, Robinson IM (1984) Cellulolytic bacteria from pig large intestine. Appl Environ Microbiol 47:219–221

    Google Scholar 

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Authors and Affiliations

  1. Department of Veterinary and Microbiological Sciences, North Dakota State University, 58105, Fargo, ND, USA

    Thomas L. Glass & Julie S. Sherwood

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  1. Thomas L. Glass
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  2. Julie S. Sherwood
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Published with the approval of the Director of the North Dakota Agricultural Experiment Station as journal article no. 2186

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Glass, T.L., Sherwood, J.S. Phosphorylation of glucose by a guanosine-5′-triphosphate (GTP)-dependent glucokinase in Fibrobacter succinogenes subsp. succinogenes S85. Arch Microbiol 162, 180–186 (1994). https://doi.org/10.1007/BF00314472

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  • DOI: https://doi.org/10.1007/BF00314472

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