The importance of ATP as a regulator of glycolytic flux in Saccharomyces cerevisiae

Larsson, Christer; Påhlman, I.-L.; Gustafsson, Lena

doi:10.1002/1097-0061(20000630)16:9<797::aid-yea553>3.3.co;2-x

Cited by 30 publications

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“…We interpret these results to mean that a minimum amount of ATP is needed internally to initiate glucose degradation. This general effect was demonstrated previously by testing the effect of different ATP concentrations on the glycolytic flux of permeabilized cells of S. cerevisiae (13). The intracellular ATP is probably required for the phosphorylation of glucose and fructose-6-phosphate by hexokinase and phosphofructokinase, respectively, both of which act in glycolysis before ATP production begins (13).…”

Section: Fig 2 Glycogen Content (Percentage Of Dry Weight) Versus Amentioning

confidence: 52%

Starvation Response ofSaccharomyces cerevisiaeGrown in Anaerobic Nitrogen- or Carbon-Limited Chemostat Cultures

Thomsson

Gustafsson

Larsson

2005

Appl Environ Microbiol

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Anaerobic starvation conditions are frequent in industrial fermentation and can affect the performance of the cells. In this study, the anaerobic carbon or nitrogen starvation response of Saccharomyces cerevisiae was investigated for cells grown in anaerobic carbon or nitrogen-limited chemostat cultures at a dilution rate of 0.1 h ؊1 at pH 3.25 or 5. Lactic or benzoic acid was present in the growth medium at different concentrations, resulting in 16 different growth conditions. At steady state, cells were harvested and then starved for either carbon or nitrogen for 24 h under anaerobic conditions. We measured fermentative capacity, glucose uptake capacity, intracellular ATP content, and reserve carbohydrates and found that the carbon, but not the nitrogen, starvation response was dependent upon the previous growth conditions. All cells subjected to nitrogen starvation retained a large portion of their initial fermentative capacity, independently of previous growth conditions. However, nitrogen-limited cells that were starved for carbon lost almost all their fermentative capacity, while carbon-limited cells managed to preserve a larger portion of their fermentative capacity during carbon starvation. There was a positive correlation between the amount of glycogen before carbon starvation and the fermentative capacity and ATP content of the cells after carbon starvation. Fermentative capacity and glucose uptake capacity were not correlated under any of the conditions tested. Thus, the successful adaptation to sudden carbon starvation requires energy and, under anaerobic conditions, fermentable endogenous resources. In an industrial setting, carbon starvation in anaerobic fermentations should be avoided to maintain a productive yeast population.

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Section: Fig 2 Glycogen Content (Percentage Of Dry Weight) Versus Amentioning

confidence: 52%

Starvation Response ofSaccharomyces cerevisiaeGrown in Anaerobic Nitrogen- or Carbon-Limited Chemostat Cultures

Thomsson

Gustafsson

Larsson

2005

Appl Environ Microbiol

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“…There was a very rapid drop in ATP content when anaerobic cells, growing exponentially, were suddenly subjected to carbon starvation. The ATP concentration after 24 h of carbon starvation could be estimated to only about 0.01 to 0.03 mM (by assuming an intracellular volume of 2 ml/g of dry weight [11]). Such levels are far below what has previously been reported to impose a limitation on glycolytic flux in S. cerevisiae (11).…”

Section: Discussionmentioning

confidence: 99%

“…On the contrary, carbon-starved cells were virtually devoid of any storage carbohydrates while both trehalose and glycogen accumulated during nitrogen starvation (16). Other factors that are important for regulation of catabolic activity are the adenine nucleotides (4,9,11). For instance, it has been shown that ATP becomes limiting for glycolytic flux at concentrations below about 1 to 1.5 mM (11) while higher ATP levels have an inhibitory effect on the rate of glycolysis (9,11).…”

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confidence: 99%

“…Other factors that are important for regulation of catabolic activity are the adenine nucleotides (4,9,11). For instance, it has been shown that ATP becomes limiting for glycolytic flux at concentrations below about 1 to 1.5 mM (11) while higher ATP levels have an inhibitory effect on the rate of glycolysis (9,11). The results obtained after addition of glucose to S. cerevisiae cells subjected to carbon or nitrogen starvation suggested that a certain concentration of ATP is required to obtain a certain glycolytic flux (16).…”

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confidence: 99%

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Carbon Starvation Can Induce Energy Deprivation and Loss of Fermentative Capacity in Saccharomyces cerevisiae

Thomsson

Larsson

Albers

et al. 2003

Appl Environ Microbiol

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Seven different strains of Saccharomyces cerevisiae were tested for the ability to maintain their fermentative capacity during 24 h of carbon or nitrogen starvation. Starvation was imposed by transferring cells, exponentially growing in anaerobic batch cultures, to a defined growth medium lacking either a carbon or a nitrogen source. After 24 h of starvation, fermentative capacity was determined by addition of glucose and measurement of the resulting ethanol production rate. The results showed that 24 h of nitrogen starvation reduced the fermentative capacity by 70 to 95%, depending on the strain. Carbon starvation, on the other hand, provoked an almost complete loss of fermentative capacity in all of the strains tested. The absence of ethanol production following carbon starvation occurred even though the cells possessed a substantial glucose transport capacity. In fact, similar uptake capacities were recorded irrespective of whether the cells had been subjected to carbon or nitrogen starvation. Instead, the loss of fermentative capacity observed in carbon-starved cells was almost surely a result of energy deprivation. Carbon starvation drastically reduced the ATP content of the cells to values well below 0.1 mol/g, while nitrogen-starved cells still contained approximately 6 mol/g after 24 h of treatment. Addition of a small amount of glucose (0.1 g/liter at a cell density of 1.0 g/liter) at the initiation of starvation or use of stationary-phase instead of log-phase cells enabled the cells to preserve their fermentative capacity also during carbon starvation. The prerequisites for successful adaptation to starvation conditions are probably gradual nutrient depletion and access to energy during the adaptation period.

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“…The catalytic activity is tightly regulated in a wide variety of organisms by diverse positive and negative effectors. The enzyme plays a major role in regulating the glycolytic flux (Hofmann, 1976;Heinrich et al, 1977;Sols, 1981;Kotlarz and Buc, 1982;Kemp and Foe, 1983;Larsson et al, 2000). In a few bacteria and most eukaryotic cells, ATP was found to function not only as a substrate but also as a potent inhibitor, whereas AMP and fructose 2,6-bis(phosphate) (Fru 2,6-P 2 ) are strong activators and are able to diminish ATP inhibition.…”

Section: Introductionmentioning

confidence: 99%

6‐phosphofructokinase from Pichia pastoris: purification, kinetic and molecular characterization of the enzyme

Kirchberger

Bär

Schellenberger

et al. 2002

Yeast

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Abstract6-Phosphofructokinase from Pichia pastoris was purified for the first time to homogeneity applying seven steps, including pseudo-affinity dye-ligand chromatography on Procion Blue H-5R-Sepharose. The specific activity of the purified enzyme was about 80 U/mg. It behaves as a typically allosteric 6-phosphofructokinase exhibiting activation by AMP and fructose 2,6-bis(phosphate), inhibition by ATP and cooperativity to fructose 6-phosphate. However, in comparison with the enzymes from Saccharomyces cerevisiae and Kluyveromyces lactis, the activation ratio of 6-phosphofructokinase from Pichia pastoris by AMP is several times higher, the ATP inhibition is stronger and the apparent affinity to fructose 6-phosphate is significantly lower. Aqueous twophase affinity partitioning with Cibacron Blue F3G-A did not reflect remarkable structural differences of the nucleotide binding sites of the Pfks from Pichia pastoris and Saccharomyces cerevisiae. The structural organisation of the active enzyme seems to be different in comparison with hetero-octameric 6-phosphofructokinases from other yeast species. The enzyme was found to be a hetero-oligomer with an molecular mass of 975 kDa (sedimentation equilibrium measurements) consisting of two distinct types of subunits in an equimolar ratio with molecular masses of 113 kDa and 98 kDa (SDS-PAGE), respectively, and a third non-covalently complexed protein component (34 kDa, SDS-PAGE). The latter seems to be necessary for the catalytic activity of the enzyme. Sequencing of the N-terminus (VTKDSIXRDLEXENXGXXFF) and of peptide fragments by applying MALDI-TOF PSD, m/z 1517.3 (DAMNVVNH) and m/z 2177.2 [AQNCNVC(L/I)SVHEAHTM] gave no relevant information about the identity of this protein.

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The importance of ATP as a regulator of glycolytic flux in Saccharomyces cerevisiae

Cited by 30 publications

References 0 publications

Starvation Response ofSaccharomyces cerevisiaeGrown in Anaerobic Nitrogen- or Carbon-Limited Chemostat Cultures

Starvation Response ofSaccharomyces cerevisiaeGrown in Anaerobic Nitrogen- or Carbon-Limited Chemostat Cultures

Carbon Starvation Can Induce Energy Deprivation and Loss of Fermentative Capacity in Saccharomyces cerevisiae

6‐phosphofructokinase from Pichia pastoris: purification, kinetic and molecular characterization of the enzyme

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