Abstract
The generation of transmembrane ion gradients by Oxalobacter formigenes cells metabolizing oxalate was studied. The magnitudes of both the transmembrane electrical potential (delta psi) and the pH gradient (internal alkaline) decreased with increasing external pH; quantitatively, the delta psi was the most important component of the proton motive force. As the extracellular pH of metabolizing cells was increased, intracellular pH increased and remained alkaline relative to the external pH, indicating that O. formigenes possesses a limited capacity to regulate internal pH. The generation of a delta psi by concentrated suspensions of O. formigenes cells was inhibited by the K+ ionophore valinomycin and the protonophore carbonyl cyanide-m-chlorophenylhydrazone, but not by the Na+ ionophore monensin. The H+ ATPase inhibitor N,N'-dicyclohexyl-carbodiimide inhibited oxalate catabolism but did not dissipate the delta psi. The results support the concept that energy from oxalate metabolism by O. formigenes is conserved not as a sodium ion gradient but rather, at least partially, as a transmembrane hydrogen ion gradient produced during the electrogenic exchange of substrate (oxalate) and product (formate) and from internal proton consumption during oxalate decarboxylation.
Full Text
The Full Text of this article is available as a PDF (292.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahmed S., Booth I. R. Quantitative measurements of the proton-motive force and its relation to steady state lactose accumulation in Escherichia coli. Biochem J. 1981 Dec 15;200(3):573–581. doi: 10.1042/bj2000573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Allison M. J., Cook H. M., Milne D. B., Gallagher S., Clayman R. V. Oxalate degradation by gastrointestinal bacteria from humans. J Nutr. 1986 Mar;116(3):455–460. doi: 10.1093/jn/116.3.455. [DOI] [PubMed] [Google Scholar]
- Allison M. J., Dawson K. A., Mayberry W. R., Foss J. G. Oxalobacter formigenes gen. nov., sp. nov.: oxalate-degrading anaerobes that inhabit the gastrointestinal tract. Arch Microbiol. 1985 Feb;141(1):1–7. doi: 10.1007/BF00446731. [DOI] [PubMed] [Google Scholar]
- Anantharam V., Allison M. J., Maloney P. C. Oxalate:formate exchange. The basis for energy coupling in Oxalobacter. J Biol Chem. 1989 May 5;264(13):7244–7250. [PubMed] [Google Scholar]
- Baetz A. L., Allison M. J. Purification and characterization of formyl-coenzyme A transferase from Oxalobacter formigenes. J Bacteriol. 1990 Jul;172(7):3537–3540. doi: 10.1128/jb.172.7.3537-3540.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baetz A. L., Allison M. J. Purification and characterization of oxalyl-coenzyme A decarboxylase from Oxalobacter formigenes. J Bacteriol. 1989 May;171(5):2605–2608. doi: 10.1128/jb.171.5.2605-2608.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Blackmore M. A., Quayle J. R. Microbial growth on oxalate by a route not involving glyoxylate carboligase. Biochem J. 1970 Jun;118(1):53–59. doi: 10.1042/bj1180053. [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]
- Bryant M. P. Commentary on the Hungate technique for culture of anaerobic bacteria. Am J Clin Nutr. 1972 Dec;25(12):1324–1328. doi: 10.1093/ajcn/25.12.1324. [DOI] [PubMed] [Google Scholar]
- Buckel W., Semmler R. A biotin-dependent sodium pump: glutaconyl-CoA decarboxylase from Acidaminococcus fermentans. FEBS Lett. 1982 Nov 1;148(1):35–38. doi: 10.1016/0014-5793(82)81237-4. [DOI] [PubMed] [Google Scholar]
- Collins M. D., Lawson P. A., Willems A., Cordoba J. J., Fernandez-Garayzabal J., Garcia P., Cai J., Hippe H., Farrow J. A. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol. 1994 Oct;44(4):812–826. doi: 10.1099/00207713-44-4-812. [DOI] [PubMed] [Google Scholar]
- Daniel S. L., Drake H. L. Oxalate- and Glyoxylate-Dependent Growth and Acetogenesis by Clostridium thermoaceticum. Appl Environ Microbiol. 1993 Sep;59(9):3062–3069. doi: 10.1128/aem.59.9.3062-3069.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daniel S. L., Hartman P. A., Allison M. J. Microbial degradation of oxalate in the gastrointestinal tracts of rats. Appl Environ Microbiol. 1987 Aug;53(8):1793–1797. doi: 10.1128/aem.53.8.1793-1797.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dawson K. A., Allison M. J., Hartman P. A. Isolation and some characteristics of anaerobic oxalate-degrading bacteria from the rumen. Appl Environ Microbiol. 1980 Oct;40(4):833–839. doi: 10.1128/aem.40.4.833-839.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dimroth P. The generation of an electrochemical gradient of sodium ions upon decarboxylation of oxaloacetate by the membrane-bound and Na+-activated oxaloacetate decarboxylase from Klebsiella aerogenes. Eur J Biochem. 1982 Jan;121(2):443–449. doi: 10.1111/j.1432-1033.1982.tb05807.x. [DOI] [PubMed] [Google Scholar]
- FitzGerald R. J., Doonan S., McKay L. L., Cogan T. M. Intracellular pH and the role of D-lactate dehydrogenase in the production of metabolic end products by Leuconostoc lactis. J Dairy Res. 1992 Aug;59(3):359–367. doi: 10.1017/s0022029900030636. [DOI] [PubMed] [Google Scholar]
- Hilpert W., Dimroth P. Conversion of the chemical energy of methylmalonyl-CoA decarboxylation into a Na+ gradient. Nature. 1982 Apr 8;296(5857):584–585. doi: 10.1038/296584a0. [DOI] [PubMed] [Google Scholar]
- Hilpert W., Schink B., Dimroth P. Life by a new decarboxylation-dependent energy conservation mechanism with Na as coupling ion. EMBO J. 1984 Aug;3(8):1665–1670. doi: 10.1002/j.1460-2075.1984.tb02030.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kamo N., Muratsugu M., Hongoh R., Kobatake Y. Membrane potential of mitochondria measured with an electrode sensitive to tetraphenyl phosphonium and relationship between proton electrochemical potential and phosphorylation potential in steady state. J Membr Biol. 1979 Aug;49(2):105–121. doi: 10.1007/BF01868720. [DOI] [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]
- Kroll R. G., Booth I. R. The role of potassium transport in the generation of a pH gradient in Escherichia coli. Biochem J. 1981 Sep 15;198(3):691–698. doi: 10.1042/bj1980691. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maloney P. C., Anantharam V., Allison M. J. Measurement of the substrate dissociation constant of a solubilized membrane carrier. Substrate stabilization of OxlT, the anion exchange protein of Oxalobacter formigenes. J Biol Chem. 1992 May 25;267(15):10531–10536. [PubMed] [Google Scholar]
- Maloney P. C. Bacterial transporters. Curr Opin Cell Biol. 1994 Aug;6(4):571–582. doi: 10.1016/0955-0674(94)90079-5. [DOI] [PubMed] [Google Scholar]
- POSTGATE J. R. A STRAIN OF DESULFOVIBRIO ABLE TO USE OXAMATE. Arch Mikrobiol. 1963 Sep 16;46:287–295. doi: 10.1007/BF00422189. [DOI] [PubMed] [Google Scholar]
- Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
- Quayle J. R., Keech D. B., Taylor G. A. Carbon assimilation by Pseudomonas oxalaticus (OXI). 4. Metabolism of oxalate in cell-free extracts of the organism grown on oxalate. Biochem J. 1961 Feb;78(2):225–236. doi: 10.1042/bj0780225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rottem S., Linker C., Wilson T. H. Proton motive force across the membrane of Mycoplasma gallisepticum and its possible role in cell volume regulation. J Bacteriol. 1981 Mar;145(3):1299–1304. doi: 10.1128/jb.145.3.1299-1304.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rottenberg H. The measurement of membrane potential and deltapH in cells, organelles, and vesicles. Methods Enzymol. 1979;55:547–569. doi: 10.1016/0076-6879(79)55066-6. [DOI] [PubMed] [Google Scholar]
- Ruan Z. S., Anantharam V., Crawford I. T., Ambudkar S. V., Rhee S. Y., Allison M. J., Maloney P. C. Identification, purification, and reconstitution of OxlT, the oxalate: formate antiport protein of Oxalobacter formigenes. J Biol Chem. 1992 May 25;267(15):10537–10543. [PubMed] [Google Scholar]
- Thauer R. K., Jungermann K., Decker K. Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev. 1977 Mar;41(1):100–180. doi: 10.1128/br.41.1.100-180.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ugurbil K., Rottenberg H., Glynn P., Shulman R. G. Phosphorus-31 nuclear magnetic resonance studies of bioenergetics in wild-type and adenosinetriphosphatase(1-) Escherichia coli cells. Biochemistry. 1982 Mar 2;21(5):1068–1075. doi: 10.1021/bi00534a038. [DOI] [PubMed] [Google Scholar]
- ten Brink B., Konings W. N. Electrochemical proton gradient and lactate concentration gradient in Streptococcus cremoris cells grown in batch culture. J Bacteriol. 1982 Nov;152(2):682–686. doi: 10.1128/jb.152.2.682-686.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]