Skip to main content
Log in

31P Nuclear magnetic resonance studies of ethanol inhibition in Zymomonas mobilis

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Ethanol inhibition of glucose catabolism in Zymomonas mobilis was investigated using 31P NMR spectroscopy in vivo and of perchloric acid extracts from cell suspensions incubated with 0, 5 and 10% (w/v) ethanol. In vivo 31P NMR experiments revealed slower glucose utilization and decreased levels of nucleoside triphosphates in the presence of 10% ethanol as compared to controls. Using 31P NMR spectroscopy of perchloric acid extracts, intracellular accumulation of 3.4 mM 3-phosphoglycerate was found when 10% ethanol was present in the medium. No accumulation of this metabolite occurred in cells incubated with 0 and 5% ethanol. Enzyme assays confirmed that phosphoglycerate-mutase and enolase were inhibited 31 and 40%, respectively, in the presence of 10% ethanol in the test system. Therefore, under the conditions used the decrease in the fermentative activity of Z. mobilis at high ethanol concentrations is due to inhibition of phosphoglycerate-mutase and enolase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

KDPG:

2-keto-3-deoxy-6-phosphogluconate

References

  • AlgarEM, ScopesRK (1985) Studies on cell-free metabolism: ethanol production by extracts of Zymomonas mobilis. J Biotechnol 2: 275–287

    Google Scholar 

  • BarrowKD, CollinsJG, NortonRS, RogersPL, SmithGM (1984) 31P Nuclear magnetic resonance studies of the fermentation of glucose to ethanol by Zymomonas mobilis. J Biol Chem 259: 5711–5716

    Google Scholar 

  • BergmeyerHU, BergmeyerJ, GrasslM (1984) Methods of enzymatic analysis, vol 2. Verlag Chemie, Weinheim, pp 182–183

    Google Scholar 

  • BradfordMM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Article  CAS  PubMed  Google Scholar 

  • BringerS, FinnRK, SahmH (1984) Effect of oxygen on the metabolism of Zumomonas mobilis. Arch Microbiol 139: 376–381

    Google Scholar 

  • CareyVC, IngramLO (1983) Lipid composition of Zymomonas mobilis: effects of ethanol and glucose. J Bacteriol 154: 1291–1300

    Google Scholar 

  • DenHollanderJA, UgurbilK, BrownTR, ShulmanRG (1981) Phosphorus-31 nuclear magnetic resonance studies of the effect of oxygen upon glycolysis in yeast. Biochemistry 20: 5871–5880

    Google Scholar 

  • FeldmannSD, SahmH, SprengerGA (1992) Pentose metabolism in Zymomonas mobilis wild type and recombinant strains. Appl Microbiol Biotechnol 38: 354–361

    Google Scholar 

  • HermansMAF, NeussB, SahmH (1991) Content and composition of hopanoids in Zymomonas mobilis under various growth conditions. J Bacteriol 173: 5592–5595

    Google Scholar 

  • IngramLO, ButtkeTM (1984) Effects of alcohols on microorganisms Adv Microb Physiol 25: 253–300

    Google Scholar 

  • KirkK, RaftosJE, KuchelPW (1986) Triethyl phosphate as an internal 31P NMR reference in biological samples. J Magn Reson 70: 484–487

    Google Scholar 

  • LeeKJ, SkotnickiML, TribeDE, RogersPL (1980) Kinetic studies on a highly productive strain of Zymomonas mobilis. Biotechnol Lett 2: 339–344

    Google Scholar 

  • Loureiro-DiasMC, SantosH (1990) Effects of ethanol on Saccharomyces cerevisiae as monitored by in vivo 31P and 31C nuclear magnetic resonance. Arch Microbiol 153: 384–391

    Google Scholar 

  • MillarDG, Griffiths-SmithK, AlgarE, ScopesRK (1982) Activity and stability of glycolytic enzymes in the presence of ethanol. Biotechnol Lett 4: 601–606

    Google Scholar 

  • NegeleinE, NollF (1984) d-Glycerate 1,3-bisphosphate. In: BergmeyerHU, BergmeyerJ, GrasslH (eds) Methods of enzymatic analysis, vol 6. Verlag Chemie, Weinheim pp 542–546

    Google Scholar 

  • OsmanYA, IngramLO (1985) Mechanism of ethanol inhibition of fermentation in Zymomonas mobilis CP4. J Bacteriol 164: 173–180

    Google Scholar 

  • OsmanYA, ConwayT, BonettiSJ, IngramLO (1987) Glycolytic flux in Zymomonas mobilis: enzyme and metabolite levels during batch fermentation. J Bacteriol 169: 3726–3736

    Google Scholar 

  • RogersPL, LeeKJ, SkotnickiML, TribeDE (1982) Ethanol production by Zymomonas mobilis. Adv Biochem Engin 23: 37–84

    Google Scholar 

  • RottenbergH (1979) The measurement of membrane potential and delta pH in cells, organelles, and vesicles. In: FleischerS, PackerL (eds) Methods in enzymology, vol 55. Academic Press, New York, pp 547–569

    Google Scholar 

  • ScopesRK, Griffiths-SmithK (1984) Use of differential dye-ligand chromatography with affinity elution for enzyme purification: 6-phosphogluconate dehydratase from Zymomonas mobilis. Anal Biochem 136: 530–534

    Google Scholar 

  • ScopesRK, TestolinV, StoterA, Griffiths-SmithK, AlgarEM (1985) Simultaneous purification and characterization of glucokinase, fructokinase and glucose-6-phosphate dehydrogenase from Zymomonas mobilis. Biochem J 228: 627–634

    Google Scholar 

  • SwingsJ, DeLeyJ (1977) The biology of Zymomonas. Bacteriol Rev 41: 1–46

    Google Scholar 

  • UgurbilK, ShulmanRG, BrownTR (1979) High-resolution 31P and 13C nuclear magnetic resonance studies of Escherichia coli cells in vivo. In: ShulmanRG (ed) Biological applications of magnetic resonance. Academic Press, New York, pp 537–589

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Strohhäcker, J., de Graaf, A.A., Schoberth, S.M. et al. 31P Nuclear magnetic resonance studies of ethanol inhibition in Zymomonas mobilis . Arch. Microbiol. 159, 484–490 (1993). https://doi.org/10.1007/BF00288598

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00288598

Key words

Navigation