TheKluyver effect for trehalose inSaccharomyces cerevisiae

Malluta, Éverson Fernando; Decker, P.; Stambuk, Boris U.

doi:10.1002/1521-4028(200007)40:3<199::aid-jobm199>3.0.co;2-e

Cited by 28 publications

(26 citation statements)

References 22 publications

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“…In addition, based on recent results on the composition of yeast extract (Zhang et al 2003), we also assumed that the media contains significant amounts of trehalose that can be used by the cells. The maximal uptake rates for the carbon sources were obtained from the literature (Strathern et al 1982;Sutherland et al 1997;Casal et al 1999;Diderich et al 1999;Malluta et al 2000), and the uptake rates for the other nutrients were set to be high enough not to be strongly growth limiting, but low enough so that the primary carbon source was still used. In addition to the primary carbon source uptake rate, the maximum oxygen uptake rate is also an important parameter in the simulations.…”

Section: In Silico Gene Deletionsmentioning

confidence: 86%

Reconstruction and Validation of Saccharomyces cerevisiae iND750, a Fully Compartmentalized Genome-Scale Metabolic Model

Duarte¹,

Herrgård²,

Palsson³

2004

Genome Res.

582

602

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A fully compartmentalized genome-scale metabolic model of Saccharomyces cerevisiae that accounts for 750 genes and their associated transcripts, proteins, and reactions has been reconstructed and validated. All of the 1149 reactions included in this in silico model are both elementally and charge balanced and have been assigned to one of eight cellular locations (extracellular space, cytosol, mitochondrion, peroxisome, nucleus, endoplasmic reticulum, Golgi apparatus, or vacuole). When in silico predictions of 4154 growth phenotypes were compared to two published large-scale gene deletion studies, an 83% agreement was found between iND750's predictions and the experimental studies. Analysis of the failure modes showed that false predictions were primarily caused by iND750's limited inclusion of cellular processes outside of metabolism. This study systematically identified inconsistencies in our knowledge of yeast metabolism that require specific further experimental investigation

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Section: In Silico Gene Deletionsmentioning

confidence: 86%

Reconstruction and Validation of Saccharomyces cerevisiae iND750, a Fully Compartmentalized Genome-Scale Metabolic Model

Duarte¹,

Herrgård²,

Palsson³

2004

Genome Res.

582

602

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“…For this reason, Malluta and coworkers (30) concluded that AGT1 is essential for trehalose assimilation probably because agt1 mutant cells were cultured in nonbuffered trehalose synthetic medium, which prevents effective Ath1p-dependent hydrolysis of trehalose. Finally, the decrease in the trehalose consumption rate of ath1 mutant cells as the sugar concentration in the medium dropped below 6 mM is consistent with the affinity constant of Agt1p for trehalose (K m ϭ 4 mM) (53).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, biochemical and genetic data were consistent with the existence of at least two trehalose transporters in S. cerevisiae: the high-affinity H ϩ -trehalose symporter Agt1p and a low-affinity transporter system whose corresponding gene remains to be characterized (19,42,(51)(52)(53)(54). While Nwaka and coworkers (36) suggested that trehalose may reach the vacuole by simple endocytosis, Malluta et al (30) proposed that the disaccharide could be taken up by Agt1p. However, to be effective, this model implies the protection of internalized trehalose against neutral trehalase and the existence of two additional transporter activities by which trehalose enters the vacuole to be degraded by Ath1p and the subsequent exit of glucose from this compartment.…”

Section: Trehalose [␣-D-glucopyranosyl-(1-1)-␣-d-glucopyranoside] Is mentioning

confidence: 99%

“…We hypothesized that the S. cerevisiae acid trehalase also is extracellular, which would give a simple alternative explanation for the mechanism of trehalose assimilation. The occurrence of trehalose transporters (30,42) prompted us to reconsider the cellular destination of trehalose molecules that are transported across the plasma membrane. In this report, we show that Ath1p is mainly localized in the periplasmic space and demonstrate that Ath1p and Agt1p belong to two independent systems for trehalose assimilation.…”

Section: Trehalose [␣-D-glucopyranosyl-(1-1)-␣-d-glucopyranoside] Is mentioning

confidence: 99%

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Two Distinct Pathways for Trehalose Assimilation in the Yeast Saccharomyces cerevisiae

Jules

Guillou

François

et al. 2004

Appl Environ Microbiol

136

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The yeast Saccharomyces cerevisiae can synthesize trehalose and also use this disaccharide as a carbon source for growth. However, the molecular mechanism by which extracellular trehalose can be transported to the vacuole and degraded by the acid trehalase Ath1p is not clear. By using an adaptation of the assay of invertase on whole cells with NaF, we showed that more than 90% of the activity of Ath1p is extracellular, splitting of the disaccharide into glucose. We also found that Agt1p-mediated trehalose transport and the hydrolysis of the disaccharide by the cytosolic neutral trehalase Nth1p are coupled and represent a second, independent pathway, although there are several constraints on this alternative route. First, the AGT1/MAL11 gene is controlled by the MAL system, and Agt1p was active in neither non-maltose-fermenting nor maltose-inducible strains. Second, Agt1p rapidly lost activity during growth on trehalose, by a mechanism similar to the sugar-induced inactivation of the maltose permease. Finally, both pathways are highly pH sensitive and effective growth on trehalose occurred only when the medium was buffered at around pH 5.0. The catabolism of trehalose was purely oxidative, and since levels of Ath1p limit the glucose flux in the cells, batch cultures on trehalose may provide a useful alternative to glucose-limited chemostat cultures for investigation of metabolic responses in yeast. Trehalose [␣-D-glucopyranosyl-(1-1)-␣-D-glucopyranoside] is a nonreducing disaccharide of glucose discovered in 1832 byWiggers in a mushroom crop and later in many other fungi, plants, and insects (13). In the yeast Saccharomyces cerevisiae, trehalose can accumulate up to 15% of the cell dry mass, depending on the growth conditions, the stage of the life cycle, or environmental stress (2, 27), and for this reason it has been considered a storage carbohydrate (15,27). Moreover, the ability of this disaccharide to protect proteins and biological membranes against adverse conditions suggests that it also plays a stress-protectant role in this yeast (49,50) and probably in other organisms that produce it (14).Intracellular levels of trehalose result from a well-controlled balance between enzymatic synthesis and degradation. In the yeast S. cerevisiae, the synthesis of trehalose is catalyzed by a UDP-glucose-dependent trehalose synthase (TPS) protein complex encoded by four genes. TPS1 and TPS2 encode trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase, respectively, while TPS3 and TSL1 code for two regulatory subunits of the TPS complex (for a review, see reference 15). Hydrolysis of trehalose into glucose can be carried out by at least two enzymes: a cytosolic or neutral trehalase encoded by NTH1 (26) and a vacuolar or acid trehalase (25, 28) encoded by ATH1 (12). The yeast genome also harbors the NTH2 gene, whose product is 77% identical to Nth1p, but until now, no trehalase activity has been associated with this protein (37). The neutral trehalase Nth1p is responsible for the intracellular mobilization ...

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“…it is Kluyver effect positive for trehalose, probably because of an insufficient level of the trehalose transporter (AGT1 product), as suggested by Malluta et al (25).…”

Section: Vol 184 2002 Sugar Transporters In Fermentative Growth Of mentioning

confidence: 99%

Respiration-Dependent Utilization of Sugars in Yeasts: a Determinant Role for Sugar Transporters

2002

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In many yeast species, including Kluyveromyces lactis, growth on certain sugars (such as galactose, raffinose, and maltose) occurs only under respiratory conditions. If respiration is blocked by inhibitors, mutation, or anaerobiosis, growth does not take place. This apparent dependence on respiration for the utilization of certain sugars has often been suspected to be associated with the mechanism of the sugar uptake step. We hypothesized that in many yeast species, the permease activities for these sugars are not sufficient to ensure the high substrate flow that is necessary for fermentative growth. By introducing additional sugar permease genes, we have obtained K. lactis strains that were capable of growing on galactose and raffinose in the absence of respiration. High dosages of both the permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. These results strongly suggest that the sugar uptake step is the major bottleneck in the fermentative assimilation of certain sugars in K. lactis and probably in many other yeasts.Kluyveromyces lactis and many other yeast species can grow on galactose and certain oligosaccharides (such as raffinose and maltose) aerobically, but they cannot grow on these sugars anaerobically or in the absence of respiration (15,18,19,35). Assimilation of these carbon sources occurs only under respiring conditions. The phenomenon has been known by the classical name of the Kluyver effect. The kind of sugars involved varies depending on the species and sometimes on the strains within a species. Although the reason for this apparent dependence on respiration for the assimilation of certain sugars is not clear, the phenomenon does appear to be brought about by the interplay of several factors involving lowered rate of transport and metabolism of certain sugars (4). Saccharomyces cerevisiae generally does not show this phenomenon (Kluyver effect negative), although K. lactis and S. cerevisiae seem to use similar pathways to metabolize galactose, raffinose, and maltose.In the present work we show that, by introducing additional Saccharomyces sugar permease genes, K. lactis cells can be released from their dependence on respiration for the assimilation of galactose and raffinose. High dosages of both permease and maltase genes were indeed necessary for K. lactis cells to grow on maltose in the absence of respiration. MATERIALS AND METHODSStrains, media, and growth conditions. K. lactis strains used in this study were PM4-4B (MAT␣ ade1 ade2 uraA), PM1-11B (MATa metA uraA lys arg rag2), P11-1D (uraA lys his2-2 lac12-230), P11-1A (lys his2-2 LAC12), 11D304 ), JBD100 (MAT␣ trp1 ura3-100 lac4-1), JBD100/M3 (MAT␣ trp1 ura3-100 lac4-1 cyt1 [16]), and JA6 (MAT␣ ade1 ade2 trp1 uraA [9]). The P11-1D lac12 mutant was obtained by sporulation of the P11 diploid constructed by crossing the original mutant 11D304 lac12 (32) with a ura3 LAC12 strain, PM1-11B. As described for the 11D304 lac12 strain (32), the lac12 mutant P11-1D is able to grow on ...

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TheKluyver effect for trehalose inSaccharomyces cerevisiae

Cited by 28 publications

References 22 publications

Reconstruction and Validation of Saccharomyces cerevisiae iND750, a Fully Compartmentalized Genome-Scale Metabolic Model

Reconstruction and Validation of Saccharomyces cerevisiae iND750, a Fully Compartmentalized Genome-Scale Metabolic Model

Two Distinct Pathways for Trehalose Assimilation in the Yeast Saccharomyces cerevisiae

Respiration-Dependent Utilization of Sugars in Yeasts: a Determinant Role for Sugar Transporters

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