Abstract
The physiology and biochemistry of Sarcina ventriculi was studied in order to determine adaptations made by the organism to changes in environmental pH. The organism altered carbon and electron flow from acetate, formate and ethanol production at neutral pH, to predominantly ethanol production at pH 3.0. Increased levels of pyruvate dehydrogenase (relative to pyruvate decarboxylase) and acetaldehyde dehydrogenase occurred when the organism was grown at neutral pH, indicating the predominance of carbon flux through the oxidative branch of the pathway for pyruvate metabolism. When the organism was grown at acid pH, there was a significant increase in pyruvate decarboxylase levels and a decrease in acetaldehyde dehydrogenase, causing flux through the non-oxidative branch of the pathway. CO2 reductase and formate dehydrogenase were not regulated as a function of growth pH. Pyruvate dehydrogenase possessed Michaelis-Menten kinetics for pyruvate with an apparent K m of 2.5 mM, whereas pyruvate decarboxylase exhibited sigmoidal kinetics, with a S0.5 of 12.0 mM. Differences in total protein banding patterns from cells grown at pH extremes suggested that synthesis of pyruvate decarboxylase and other enzymes was in part responsible for metabolic regulation of the fermentation products formed.
Similar content being viewed by others
References
Andersch W, Bahl H, Gottschalk G (1983) Level of enzymes involved in acetate, butyrate, acetone and butanol formation by Clostridium acetobutylicum. Eur J Appl Biotechnol 18: 327–332
Bahl H, Andersch W, Gottschalk G (1982) Continuous production of acetone and butanol by Clostridium acetobutylicum grown in a two-stage phosphate limited chemostat. Eur J Appl Microbiol Biotech 15: 201–205
Baronofsky JJ, Schreurs WJA, Kashket ER (1984) Uncoupling by acetic acid limits growth of and acetogenesis by Clostridium thermoaceticum. Appl Environ Microbiol 48: 1134–1139
Booth IR (1985) Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49: 359–378
Canale-Parola E (1986) Genus Sarcina Goodsir 1842, 434AL. In: Sneath PH, Mair NS, Sharpe ME, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 2. Williams & Wilkins, Baltimore, pp 1100–1103
Goodwin S, Zeikus JG (1987) Physiological adaptations of anaerobic bacteria to low pH: metabolic control of proton motive force in Sarcina ventriculi. J Bacteriol 169: 2150–2157
Gottschal JC, Morris JG (1981) The induction of acetone and butanol production in cultures of Clostridium acetobutylicum by elevated concentrations of acetate and butyrate. FEMS Microbiol Lett 12: 385–389
Gottwald M, Gottschalk G (1985) The internal pH of Clostridium acetobutylicum and its effect on the shift from acid to solvent formation. Arch Microbiol 143: 42–46
Hartmanis MGN, Gatenbeck S (1984) Intermediary metabolism in Clostridium acetobutylicum: Levels of enzymes involved in the formation of acetate and butyrate. Appl Environ Microbiol 47: 1277–1283
Hopner T, Knappe J (1974) Formate determination with formate dehydrogenase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York, pp 1551–1555
Huang L, Forsberg CW, Gibbins LN (1986) Influence of external pH and fermentation products on Clostridium acetobutylicum intracellular pH and cellular distribution of fermentation products. Appl Environ Microbiol 51: 1230–1234
Kell DB, Peck MW, Rodger G, Morris JG (1981) On the permeability to weak acids and bases of the cytoplasmic membrane of Clostridium pasteurianum. Biochem Biophys Res Commun 99: 81–88
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Lamed R, Zeikus JG (1980) Ethanol production by thermophilic bacteria: relationship between fermentation product yields of and catabolic enzyme activities in Clostridium thermocellum and Thermoanaerobium brockii. J Bacteriol 144: 569–578
LaPorte DC, Walsh K, Koshland DE (1984) The branch point effect. Ultrasensitivity and subsensitivity to metabolic control. J Biol Chem 259: 14068–14075
Lowe S, Pankratz HS, Zeikus JG (1989) Influence of pH extremes on sporulation and ultrastructure of Sarcina ventriculi. J Bacteriol 171: 3775–3781
Melville SB, Michel TA, Macy JM (1988) Regulation of carbon flow in Selenomonas ruminantium grown in glucose-limited continuous culture. J Bacteriol 170: 5305–5311
Needels DL, Wilson JE (1983) The identity of hexokinase activities from mitochondrial and cytoplasmic fractions of rat brain homogenates. J Neurochem 40: 1134–1143
Rengpipat S, Lowe SE, Zeikus JG (1988) Effect of salt concentrations on the physiology and biochemistry of Halobacteroides acetoethylicus. J Bacteriol 170: 3065–3071
Riebeling V, Thauer RK, Jungermann K (1975) The internal-alkaline pH gradient, sensitive to uncoupler and ATPase inhibitor, in growing Clostridium pasteurianum. Eur J Biochem 55: 445–453
Stephenson MP, Dawes EA (1971) Pyruvic acid and formic acid metabolism in Sarcina ventriculi and the role of ferredoxin. J Gen Microbiol 69: 331–343
Tilak Kvbr (1970) Studies on Sarcina (Zymosarcina) ventriculi. Sci Cult 36: 399–400
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Lowe, S.E., Zeikus, J.G. Metabolic regulation of carbon and electron flow as a function of pH during growth of Sarcina ventriculi . Arch. Microbiol. 155, 325–329 (1991). https://doi.org/10.1007/BF00243450
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00243450