Abstract
We investigated the intracellular physiological conditions associated with the induction of butanol-producing enzymes in Clostridium acetobutylicum. During the acidogenic phase of growth, the internal pH decreased in parallel with the decrease in the external pH, but the internal pH did not go below 5.5 throughout batch growth. Butanol was found to dissipate the proton motive force of fermenting C. acetobutylicum cells by decreasing the transmembrane pH gradient, whereas the membrane potential was affected only slightly. In growing cells, the switch from acid to solvent production occurred when the internal undissociated butyric acid concentration reached 13 mM and the total intracellular undissociated acid concentration (acetic plus butyric acids) was at least 40 to 45 mM. Similar values were obtained when cultures were supplemented with 50 mM butyric acid initially or when a phosphate-buffered medium was used instead of an acetate-buffered medium. To measure the induction of the enzymes involved in solvent synthesis, we determined the rates of conversion of butyrate to butanol in growing cells. The rate of butanol formation reached a maximum in the mid-solvent phase, when the butanol concentration was 50 mM. Although more solvent accumulated later, de novo enzyme synthesis decreased and then ceased.
Full text
PDF





Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baronofsky J. J., Schreurs W. J., Kashket E. R. Uncoupling by Acetic Acid Limits Growth of and Acetogenesis by Clostridium thermoaceticum. Appl Environ Microbiol. 1984 Dec;48(6):1134–1139. doi: 10.1128/aem.48.6.1134-1139.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Booth I. R. Regulation of cytoplasmic pH in bacteria. Microbiol Rev. 1985 Dec;49(4):359–378. doi: 10.1128/mr.49.4.359-378.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowles L. K., Ellefson W. L. Effects of butanol on Clostridium acetobutylicum. Appl Environ Microbiol. 1985 Nov;50(5):1165–1170. doi: 10.1128/aem.50.5.1165-1170.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Datta R., Zeikus J. G. Modulation of acetone-butanol-ethanol fermentation by carbon monoxide and organic acids. Appl Environ Microbiol. 1985 Mar;49(3):522–529. doi: 10.1128/aem.49.3.522-529.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davies R., Stephenson M. Studies on the acetone-butyl alcohol fermentation: Nutritional and other factors involved in the preparation of active suspensions of Cl. acetobutylicum (Weizmann). Biochem J. 1941 Dec;35(12):1320–1331. doi: 10.1042/bj0351320. [DOI] [PMC free article] [PubMed] [Google Scholar]
- George H. A., Chen J. S. Acidic Conditions Are Not Obligatory for Onset of Butanol Formation by Clostridium beijerinckii (Synonym, C. butylicum). Appl Environ Microbiol. 1983 Aug;46(2):321–327. doi: 10.1128/aem.46.2.321-327.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gottschalk G., Bahl H. Feasible improvements of the butanol production by Clostridium acetobutylicum. Basic Life Sci. 1981;18:463–471. doi: 10.1007/978-1-4684-3980-9_27. [DOI] [PubMed] [Google Scholar]
- Gottwald M., Hippe H., Gottschalk G. Formation of n-Butanol from d-Glucose by Strains of the "Clostridium tetanomorphum" Group. Appl Environ Microbiol. 1984 Sep;48(3):573–576. doi: 10.1128/aem.48.3.573-576.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harold F. M. Conservation and transformation of energy by bacterial membranes. Bacteriol Rev. 1972 Jun;36(2):172–230. doi: 10.1128/br.36.2.172-230.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartmanis M. G., Gatenbeck S. Intermediary Metabolism in Clostridium acetobutylicum: Levels of Enzymes Involved in the Formation of Acetate and Butyrate. Appl Environ Microbiol. 1984 Jun;47(6):1277–1283. doi: 10.1128/aem.47.6.1277-1283.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herrero A. A., Gomez R. F., Roberts M. F. 31P NMR studies of Clostridium thermocellum. Mechanism of end product inhibition by ethanol. J Biol Chem. 1985 Jun 25;260(12):7442–7451. [PubMed] [Google Scholar]
- Holt R. A., Stephens G. M., Morris J. G. Production of Solvents by Clostridium acetobutylicum Cultures Maintained at Neutral pH. Appl Environ Microbiol. 1984 Dec;48(6):1166–1170. doi: 10.1128/aem.48.6.1166-1170.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huang L., Gibbins L. N., Forsberg C. W. Transmembrane pH gradient and membrane potential in Clostridium acetobutylicum during growth under acetogenic and solventogenic conditions. Appl Environ Microbiol. 1985 Oct;50(4):1043–1047. doi: 10.1128/aem.50.4.1043-1047.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hutkins R. W., Kashket E. R. Phosphotransferase Activity in Clostridium acetobutylicum from Acidogenic and Solventogenic Phases of Growth. Appl Environ Microbiol. 1986 May;51(5):1121–1123. doi: 10.1128/aem.51.5.1121-1123.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ingram L. O., Buttke T. M. Effects of alcohols on micro-organisms. Adv Microb Physiol. 1984;25:253–300. doi: 10.1016/s0065-2911(08)60294-5. [DOI] [PubMed] [Google Scholar]
- Jones D. T., van der Westhuizen A., Long S., Allcock E. R., Reid S. J., Woods D. R. Solvent Production and Morphological Changes in Clostridium acetobutylicum. Appl Environ Microbiol. 1982 Jun;43(6):1434–1439. doi: 10.1128/aem.43.6.1434-1439.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kashket E. R., Barker S. L. Effects of potassium ions on the electrical and pH gradients across the membrane of Streptococcus lactis cells. J Bacteriol. 1977 Jun;130(3):1017–1023. doi: 10.1128/jb.130.3.1017-1023.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kashket E. R. The proton motive force in bacteria: a critical assessment of methods. Annu Rev Microbiol. 1985;39:219–242. doi: 10.1146/annurev.mi.39.100185.001251. [DOI] [PubMed] [Google Scholar]
- Kim B. H., Bellows P., Datta R., Zeikus J. G. 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. 1984 Oct;48(4):764–770. doi: 10.1128/aem.48.4.764-770.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Largier S. T., Long S., Santangelo J. D., Jones D. T., Woods D. R. Immobilized Clostridium acetobutylicum P262 Mutants for Solvent Production. Appl Environ Microbiol. 1985 Aug;50(2):477–481. doi: 10.1128/aem.50.2.477-481.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MITCHELL P. Coupling of phosphorylation to electron and hydrogen transfer by a chemi-osmotic type of mechanism. Nature. 1961 Jul 8;191:144–148. doi: 10.1038/191144a0. [DOI] [PubMed] [Google Scholar]
- Mitchell P. Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. Biol Rev Camb Philos Soc. 1966 Aug;41(3):445–502. doi: 10.1111/j.1469-185x.1966.tb01501.x. [DOI] [PubMed] [Google Scholar]
- Monot F., Martin J. R., Petitdemange H., Gay R. Acetone and Butanol Production by Clostridium acetobutylicum in a Synthetic Medium. Appl Environ Microbiol. 1982 Dec;44(6):1318–1324. doi: 10.1128/aem.44.6.1318-1324.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osman Y. A., Ingram L. O. Mechanism of ethanol inhibition of fermentation in Zymomonas mobilis CP4. J Bacteriol. 1985 Oct;164(1):173–180. doi: 10.1128/jb.164.1.173-180.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paterson S. J., Butler K. W., Huang P., Labelle J., Smith I. C., Schneider H. The effects of alcohols on lipid bilayers: a spin label study. Biochim Biophys Acta. 1972 Jun 20;266(3):597–602. doi: 10.1016/0006-3002(72)90003-0. [DOI] [PubMed] [Google Scholar]
- ROSS D. The acetone-butanol fermentation. Prog Ind Microbiol. 1961;3:71–90. [PubMed] [Google Scholar]
- Riebeling V., Thauer R. K., Jungermann K. The internal-alkaline pH gradient, sensitive to uncoupler and ATPase inhibitor, in growing Clostridium pasteurianum. Eur J Biochem. 1975 Jul 1;55(2):445–453. doi: 10.1111/j.1432-1033.1975.tb02181.x. [DOI] [PubMed] [Google Scholar]
- Zeikus J. G. Chemical and fuel production by anaerobic bacteria. Annu Rev Microbiol. 1980;34:423–464. doi: 10.1146/annurev.mi.34.100180.002231. [DOI] [PubMed] [Google Scholar]