Effect of the pH of the incubation medium on glycolysis and respiration in Saccharomyces cerevisiae

Peña, Antonio; Cinco, G.; Gómez‐Puyou, Armando; Tuena, M.

doi:10.1016/0003-9861(72)90359-1

Cited by 86 publications

(33 citation statements)

References 28 publications

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“…We introduced the fluorescent dye BCECF-AM, which measures vacuolar pH in yeast (11,30), or yeast pHluorin, a green fluorescent protein analog that measures cytosolic pH (31), into wild-type and vma mutant cells and studied the dynamic adjustments of cytosolic and vacuolar pH in response to glucose and KCl addition. As described above, both glucose and potassium ion are strongly implicated in cytosolic and vacuolar pH homeostasis (25,26,32,48). Fig.…”

Section: Resultsmentioning

confidence: 83%

“…Potassium ion is primarily responsible for balancing the plasma membrane potential in yeast (24). Transport of K ϩ ion into the cell results in a depolarization of the plasma membrane, allowing stimulation of Pma1p (22) and consequent cytosolic alkalinization (25,26), and pH-responsive regulation of the Trk1p potassium transporter via phosphorylation provides additional means of activating Pma1p when cytosolic pH drops. The exact mechanisms for balancing the membrane potential at the vacuolar membrane are less clear; many anion channels and other transporters are present in the vacuolar membrane that could contribute to the final potential, but their individual contributions to membrane potential are not well defined.…”

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confidence: 99%

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Vacuolar and Plasma Membrane Proton Pumps Collaborate to Achieve Cytosolic pH Homeostasis in Yeast

Martínez-Muñoz¹,

Kane

2008

Journal of Biological Chemistry

239

238

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Vacuolar proton-translocating ATPases (V-ATPases) play a central role in organelle acidification in all eukaryotic cells. To address the role of the yeast V-ATPase in vacuolar and cytosolic pH homeostasis, ratiometric pH-sensitive fluorophores specific for the vacuole or cytosol were introduced into wild-type cells and vma mutants, which lack V-ATPase subunits. Transiently glucose-deprived wild-type cells respond to glucose addition with vacuolar acidification and cytosolic alkalinization, and subsequent addition of K ؉ ion increases the pH of both the vacuole and cytosol. In contrast, glucose addition results in an increase in vacuolar pH in both vma mutants and wild-type cells treated with the V-ATPase inhibitor concanamycin A. Cytosolic pH homeostasis is also significantly perturbed in the vma mutants. Even at extracellular pH 5, conditions optimal for their growth, cytosolic pH was much lower, and response to glucose was smaller in the mutants. In plasma membrane fractions from the vma mutants, activity of the plasma membrane proton pump, Pma1p, was 65-75% lower than in fractions from wildtype cells. Immunofluorescence microscopy confirmed decreased levels of plasma membrane Pma1p and increased Pma1p at the vacuole and other compartments in the mutants. Pma1p was not mislocalized in concanamycin-treated cells, but a significant reduction in cytosolic pH under all conditions was still observed. We propose that short-term, V-ATPase activity is essential for both vacuolar acidification in response to glucose metabolism and for efficient cytosolic pH homeostasis, and long-term, V-ATPases are important for stable localization of Pma1p at the plasma membrane.The importance of V-ATPases 3 for acidification of the vacuole/lysosomes, Golgi apparatus, and endosomes of eukaryotic cells is well established (1, 2). Multiple cellular processes, including secondary transport of ions and metabolites, maturation of iron transporters, endocytic and biosynthetic protein sorting, and zymogen activation depend on compartment acidification and have been linked to V-ATPase activity (1, 3). In some cells such as macrophages, V-ATPases play specialized roles that clearly include regulation of cytosolic pH (4, 5). However, although V-ATPases pump protons from the cytosol into organelles in all cells, they are not generally believed to play a major role in cytosolic pH regulation.The yeast Saccharomyces cerevisiae has emerged as a major model system for eukaryotic V-ATPases. One reason for this is that yeast mutants lacking all V-ATPase activity (vma mutants) are viable, but loss of V-ATPase activity in eukaryotes other than fungi is lethal (6 -9). Yeast vma mutants do exhibit a set of distinctive phenotypes, however, that includes the inability to grow at pH values lower than 3 or higher than 7 and sensitivity to high extracellular calcium concentrations (2). This Vma Ϫ phenotype suggests a perturbation of pH homeostasis in these cells that is not fully understood. It has been suggested that vma mutants survive at low extracellular p...

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Section: Resultsmentioning

confidence: 83%

mentioning

confidence: 99%

Vacuolar and Plasma Membrane Proton Pumps Collaborate to Achieve Cytosolic pH Homeostasis in Yeast

Martínez-Muñoz¹,

Kane

2008

Journal of Biological Chemistry

239

238

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“…50% inhibition of the fluorescence change at 50 M and complete inhibition at 200 M (Fig. 3A); a very much smaller effect of ethanol on fluorescence (ethanol is known to be a poor substrate for energizing transport [13,14]) was completely blocked by PClP (Fig. 3B).…”

Section: Resultsmentioning

confidence: 96%

Proton pumping and the internal pH of yeast cells, measured with pyranine introduced by electroporation

et al. 1995

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The measurement of internal pH in microorganisms, in yeast cells and in cells in general, has been studied for many years. Several mechanisms are involved in the regulation of the internal pH of the cell, many cellular processes are regulated by the internal pH, and many transport processes depend on the H ϩ cycle. In yeast cells, very crude procedures were initially used, with disruption of the cells by boiling or freezing and thawing, after which the pH of the resulting sap was measured. In 1950, the group of Conway used the distribution of weak acids, such as carbonic or propionic acid, to measure the internal pH of yeast cells (5,6). By these methods, the pH was probably obtained as an average of that of the entire cell interior, including all internal compartments. More recently, other methods have been used; among them, the shift of the P i peak in nuclear magnetic resonance spectra has been useful but complicated and expensive (1, 3).In yeast cells, the use of ionizable fluorescent probes capable of crossing the membrane and distributing between the cells, organelles, or vesicles depending on the internal and external pH (16) has proven useless. Slavík (17) first introduced indicators into yeast cells whose fluorescence depends on the surrounding pH; some of these, which are available commercially, can be introduced into the cells as acetoxymethyl esters (the permeant form), which are cleaved inside by esterases, transforming them into an impermeant form and preventing their efflux. A recent report on the use of one of these dyes to measure the internal pH of yeast cells has appeared (9). However, Slayman et al. (18) have pointed out some of the drawbacks of these dyes; because of the concentration of esterases in some intracellular compartments, they are preferentially hydrolyzed and accumulated in vacuoles or other internal compartments which accumulate hydrolytic enzymes.Kano and Fendler (10) introduced the use of pyranine (8-hydroxy-1,3,6-pyrene-trisulfonic acid), a fluorescent dye with a ionizable -OH group, which shows remarkable pH dependence in fluorescence. The dye could be trapped in liposomes and used to estimate their internal pH (see also reference 4), the most important advantage being the relatively low interaction with the bilayer or proteins, because of its hydrophilic nature.Unfortunately, this property makes its use with whole cells difficult, because of problems with entry. However, a controlled electric shock of high intensity and short duration appears to produce transient openings of the cell membrane that are closed in a short time after the treatment (11). This phenomenon has been used to introduce even substances with high molecular weights into cells by a procedure known as electroporation. This method has been successfully used in yeast cells, and great improvements have been made recently (2,8).The present communication deals with the use of pyranine to measure the internal pH of yeast cells. MATERIALS AND METHODSCells from an isolated colony of commercial yeast cells (La Azt...

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“…Results of both methods were 97-100 % identical. Such promising identically have urged the author of this study to depend on the statistical analysis of those great scientists (Man et al 2011;Xin et al 2003;Phisalaphong et al 2005;Kadambiniguar 2006;Pena et al 1972;Diaz-Ricci et al 1992;Hill et al 1993;Douka 1999;Sprenger 1996;Moawad 2010;Moawad 2011a, b;Moawad 2012a, b;Austriaco 1996;Kennedy 1994;Voet 1586;Tang et al 2008;Bai et al 2007;Yadav et al 1997;Le Man et al 2008;Nahvi et al 2002;Le Man et al 2010;APHA 1998;Miller 1959;Lang et al 2005;Garda et al 2005) on purpose, regarding their results as a ''gold standard'' or fully accepted measurement against which to make a comparison to determine accuracy of the approach.…”

Section: Discussionmentioning

confidence: 99%

“…Phisalaphong et al developed a mathematical model to describe the effects of temperature on the kinetic parameters of ethanol fermentation by the flocculating yeast, Saccharomyces cerevisiae M30, and using cane molasses as the substrate from the beginning up to the stationary phase (Phisalaphong et al 2005). Also, effects of pH values of 5.0, 6.0, 7.0 and 8.0 on fermentation were investigated in prior studies concluded that low pH inhibits the yeast multiplication while the inhibitory effect of pH (at the high level) on the ethanol yield could be due to the lower ATP production during the metabolic changes in S. cerevisiae (Kadambiniguar 2006;Pena et al 1972). With respect to microorganisms, an understanding of its physiological characteristics including factors that regulate carbon and energy metabolism during growth can furnish useful information when engineering a bioconversion process involving different substrates (DiazRicci et al 1992;Hill et al 1993).…”

Section: Introductionmentioning

confidence: 99%

Optimizing bioethanol production by regulating yeast growth energy

Moawad

2012

Syst Synth Biol

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The goal of this work is to optimize production of bio-ethanol by fermentation through regulating yeast growth energy (YGE), and provide the mechanism of ethanol production from food-waste leachate (FWL) using yeast (S. cerevisiae) as inoculums to be predictable and controllable. The wide range of reduced sugar concentration (RSC) which is commonly administered from low (35 g per liter) to very high (100 g per liter) is responsible for costs increasing besides risks of FWL contamination and death of yeast cells. A mathematical model is presented to describe yeast growth energy (YGE) due to RSC doses along with predicting the amounts of ethanol yield by each dose to identify the optimum one. Simulations of the presented model showed that YGE, energy intake (EI), and their produced ethanol energy (PEE) are always balanced during fermentation process according to the law of conservation of energy. For a better fermentation rate in a continuous process and a large-scale production; YGE should be less than half of EI and more than its quarter (i.e. 1 4 EI YGE 1 2 EI) which keeps the residual energy less than YGE to avoid risks of osmotic stresses or aging of cells allowing the survival of all yeast cells as long as possible to maximize ethanol production and decrease productivity costs.

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Effect of the pH of the incubation medium on glycolysis and respiration in Saccharomyces cerevisiae

Cited by 86 publications

References 28 publications

Vacuolar and Plasma Membrane Proton Pumps Collaborate to Achieve Cytosolic pH Homeostasis in Yeast

Vacuolar and Plasma Membrane Proton Pumps Collaborate to Achieve Cytosolic pH Homeostasis in Yeast

Proton pumping and the internal pH of yeast cells, measured with pyranine introduced by electroporation

Optimizing bioethanol production by regulating yeast growth energy

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