Activity of the plasma membrane H(+)-ATPase and optimal glycolytic flux are required for rapid adaptation and growth of Saccharomyces cerevisiae in the presence of the weak-acid preservative sorbic acid

Abstract

The weak acid sorbic acid transiently inhibited the growth of Saccharomyces cerevisiae in media at low pH. During a lag period, the length of which depended on the severity of this weak-acid stress, yeast cells appeared to adapt to this stress, eventually recovering and growing normally. This adaptation to weak-acid stress was not due to metabolism and removal of the sorbic acid. A pma1-205 mutant, with about half the normal membrane H ؉-ATPase activity, was shown to be more sensitive to sorbic acid than its parent. Sorbic acid appeared to stimulate plasma membrane H ؉-ATPase activity in both PMA1 and pma1-205. Consistent with this, cellular ATP levels showed drastic reductions, the extent of which depended on the severity of weak-acid stress. The weak acid did not appear to affect the synthesis of ATP because CO 2 production and O 2 consumption were not affected significantly in PMA1 and pma1-205 cells. However, a glycolytic mutant, with about one-third the normal pyruvate kinase and phosphofructokinase activity and hence a reduced capacity to generate ATP, was more sensitive to sorbic acid than its isogenic parent. These data are consistent with the idea that adaptation by yeast cells to sorbic acid is dependent on (i) the restoration of internal pH via the export of protons by the membrane H ؉-ATPase in an energy-demanding process and (ii) the generation of sufficient ATP to drive this process and still allow growth.

Transport of carboxylic acids in yeasts

“…Upon exposure to weak acids, at low pH, S. cerevisiae cells display an adaptative stress response, which involves both the activation of the plasma membrane H 1 -ATPase, regulating the intracellular pH, pH in and homeostasis (Viegas & Sá-Correia, 1991;Holyoak et al, 1996;Piper et al, 1997;Viegas et al, 1998), and the induction of Pdr12, a plasma membrane carboxylate efflux pump (Piper et al, 1997(Piper et al, , 1998. This ATP-binding cassette (ABC) transporter is a member of the Pleiotropic Drug Resistance (PDR) Subfamily (TC 3.A.1.205.3).…”

“…The acid anion cannot readily diffuse out of the cell and accumulates. The protons released acidify the cytoplasm and can alter several metabolic pathways (Krebs et al, 1983;Pampulha & Loureiro-Dias, 1990Holyoak et al, 1996). Additionally, the intracellular protons can also influence free radical production, leading to severe oxidative stress, which is a major component of weak organic acid stress found in the yeast Saccharomyces cerevisiae under aerobic conditions (Piper et al, 2001).…”

The RIM101 pathway has a role inSaccharomyces cerevisiaeadaptive response and resistance to propionic acid and other weak acids

“…The dissociation in the near-neutral cytoplasm of the permeant acid form leads to an additional acidification of the cell interior and to the accumulation of the counter ion. The acidification of the cytosol may lead to the inhibition of critical metabolic pathways (Krebs et al, 1983;Holyoak et al, 1997). The membrane effects of weak acids are particularly deleterious in the case of the more lipophilic compounds (Piper et al, 2001;Teixeira et al, 2007).…”

Unraveling the role of membrane microdomains during microbial infections

“…The polymeric compound chitosan exerts its fungicial effect by permeabilizing plasma membrane of filamentous fungi (Palma-Guerrero et al 2010). Additionally, the role of detergent-resistant membranes in fungi for concentrating VFs-phospholipase B1 and Cu/ Zn superoxide dismutase; and proton pump protein Pma1 was demonstrated by Siafakas et al and Holyoak groups, respectively (Siafakas et al 2006;Holyoak et al 1996). Several reports have demonstrated the functional role of these VFs in evading hosts immune responses and Pma1 in the pathogenicity of C. neoformans (Farnoud et al 2014).…”