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Increased levels of ATP and NADH are associated with increased solvent production in continuous cultures of Clostridium acetobutylicum

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Summary

Levels of intracellular NAD, NADH, ADP, and ATP were measured in batch and continuous cultures of Clostridium acetobutylicum. ATP levels during glucose-sufficient (non-glucose limited), solvent-producing steady states were five to eight times as high as in glucose-limited (acidogenic) steady state cultures (4.8 μmole/g versus 0.9 μmole/g). NADH was also higher in glucose-sufficient cultures, although the increase was not as pronounced as with ATP. When glucose-limited cultures were sparged with CO, the resulting shift to solvent production was accompanied by three- or four-fold increases in NADH and ATP levels. In batch cultures, NADH and ATP levels were relatively high at all times compared to levels in continuous cultures. Although the levels of these nucleotides showed systematic trends during normal batch fermentations, there was no significant change with the onset of solvent production.

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References

  • Afschar AS, Biebl H, Schaller K, Schügerl K (1985) Production of acetone and butanol by Clostridium acetobutylicum in continuous culture with cell recycle. Appl. Microbiol Biotechnol 22:394–398

    Google Scholar 

  • Bahl H, Andersch W, Braun K, Gottschalk G (1982a) Effect of pH and butyrate concentration on the production of acetone and butanol by Clostridium acetobutylicum grown in continuous culture. Eur J Appl Microbiol Biotechnol 14:17–20

    Google Scholar 

  • Bahl H, Andersch W, Gottschalk G (1982b) Continuous production of acetone and butanol by Clostridium acetobutylicum in a two-stage phosphate limited chemostat. Eur J Appl Microbiol Biotechnol 15:201–205

    Google Scholar 

  • Bahl H, Gottwald M, Kuhn A, Rale V, Andersch W Gottschalk G (1986) Nutritional factors affecting the ratio of solvents produced by Clostridium acetobutylicum. Appl Environ Microbiol 52:169–172

    Google Scholar 

  • Bernofsky C, Swan M (1973) An improved cycling assay for nicotinamide adenine dinucleotide. Anal Biochem 53:452–458

    Google Scholar 

  • Cole HA, Wimpenny JW, Hughes DE (1967) The ATP pool in Escherichia coli. I. Measurement of the pool using a modified luciferase assay. Biochem Biophys Acta, 143:445–453

    Google Scholar 

  • Datta R, Zeikus JG (1985) Modulation of acetone-butanolethanol fermentation by carbon monoxide and organic acids. Appl Environ Microbiol 49:522–529

    Google Scholar 

  • Dürre P, Kuhn A, Gottwald M, Gottschalk G (1987) Enzymatic investigations on butanol dehydrogenase and butyraldehyde dehydrogenase in extracts of Clostridium acetobutylicum. Appl Microbiol Biotechnol 26:268–272

    Google Scholar 

  • Ferras E, Minier M, Goma G (1986) Acetonobutylic fermentation: Improvement of performances by coupling continuous fermentation and ultrafiltration. Biotechnol Bioeng 28:523–533

    Google Scholar 

  • Hüsemann MHW, Papoutsakis ET (1988) Solventogenesis in Clostridium acetobutylicum fermentations related to carboxylic acid and proton concentrations. Biotechnol Bioeng 32:843–852

    Google Scholar 

  • Hüsemann MHW, Papoutsakis ET (1989) Enzyme levels and the intracellular pH in batch and continuous fermentations of Clostridium acetobutylicum. Appl Microbiol Biotechnol 30:585–595

    Google Scholar 

  • Kim BH, Bellows P, Datta R, Zeikus JG, (1984) Control of carbon and electron flow in Clostridium acetobutylicum fermentations: Utilization of carbon monoxide to inhibit hydrogen production and to enhance butanol yields. Appl Environ Microbiol 48:764–770

    Google Scholar 

  • London J, Knight M (1966) Concentrations of nicotinamide nucleotide coenzymes in micro-organisms. J Gen Microbiol 44:241–254

    Google Scholar 

  • McLaughlin JK, Meyer CL, Papoutsakis ET (1984) Gas chromatography and gateway sensors for on-line state estimation of complex fermentations (butanol/acetone fermentation). Biotechnol Bioeng 27:1246–1257

    Google Scholar 

  • Meyer CL, McLaughlin JK, Papoutsakis ET (1985a) The effect of CO on growth and product formation in batch cultures of Clostridium acetobutylicum. Biotechnol Lett 7:37–42

    Google Scholar 

  • Meyer CL, McLaughlin JK, Papoutsakis ET (1985b) On-line chromatographic analysis and fermentor state characterization of butanol/acetone fermentations. Ann NY Acad Sci 469:350–363

    Google Scholar 

  • Meyer CL, Roos JW, Papoutsakis ET (1986) Carbon monoxide gasing leads to alcohol production and butyrate uptake without acetone formation in continuous cultures of Clostridium acetobutylicum. Appl Microbiol Biotechnol 24:159–167

    Google Scholar 

  • Meyer CL, Papoutsakis ET (1989a) Continuous and biomass recycle fermentations of Clostridium acetobutylicum. I. ATP supply and demand determines product selectivity. Bioproc Eng (in press)

  • Meyer CL, Papoutsakis ET (1989b) Continuous and biomass recycle fermentations of Clostridium acetobutylicum. II. Novel patterns in energetics and product-formation kinetics. Bioproc Eng (in press)

  • Monot F, Engasser JM, Petitdemange H (1983) Regulation of acetone butanol production in batch and continuous cultures of Clostridium acetobutylicum. Biotechnol Bioeng Symp 13:207–216

    Google Scholar 

  • Papoutsakis ET (1983) A useful equation for fermentations of butyric acid bacteria. In: Bu'Lock JD, Bu'Lock AJ (eds) btThe Acetone-butanol fermentation and related topics. Science and Technology Letters, Kew, Surrey, U.K., pp 121–126

  • Papoutsakis ET (1984) Equations and calculations for fermentations of butyric acid bacteria. Biotechnol Bioeng 26:174–187

    Google Scholar 

  • Pierrot P, Fick M, Engasser JM (1986) Continuous acetonebutanol fermentation with high productivity by cell ultrafiltration and recycling. Biotechnol Lett 8:253–256

    Google Scholar 

  • Roos JW, McLaughlin JK, Papoutsakis ET (1985) The effect of pH on nitrogen supply, cell lysis and solvent production in fermentations of Clostridium acetobutylicum. Biotechnol Bioeng 27:681–694

    Google Scholar 

  • Schlote D, Gottschalk G (1986) Effect of cell recycle on continuous butanol-acetone fermentation with Clostridium acetobutylicum under phosphate limitation. Appl Microbiol Biotechnol 24:1–5

    Google Scholar 

  • Terracciano JS, Kashket ER (1986) Intracellular conditions required for initiation of solvent production by Clostridium acetobutylicum. Appl Environ Microbiol 52:86–91

    Google Scholar 

  • Wimpenny JW, Firth A (1972) Levels of nicotinamide adenine dinucleotide and reduced nicotinamide adenine dinucleotide in facultative bacteria and the effect of oxygen. J Bacteriol 111:24–32

    Google Scholar 

  • Wulff K, Doppen W (1985) ATP: Luminometric method. In: Bergmeyer HU (ed) Methods of enzymatic analysis (3rd edn). Volume VII, VCH Publishers, Weinheim, pp 357–364

    Google Scholar 

  • Yan R, Zhu C, Golemboski C, Chen J (1988) Expression of solvent-forming enzymes and onset of solvent production in batch cultures of Clostridium beijerinckii (“Clostridium butylicum”). Appl Environ Microbiol 54:642–648

    Google Scholar 

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Meyer, C.L., Papoutsakis, E.T. Increased levels of ATP and NADH are associated with increased solvent production in continuous cultures of Clostridium acetobutylicum . Appl Microbiol Biotechnol 30, 450–459 (1989). https://doi.org/10.1007/BF00263849

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  • DOI: https://doi.org/10.1007/BF00263849

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