Two distinct pathways for trehalose assimilation in the yeast Saccharomyces cerevisiae.
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.
Jules
Matthieu
M
Centre de Bioingénierie Gilbert Durand, UMR-CNRS 5504, UMR-INRA 792, Complexe Scientifique de Rangueil, 31077 Toulouse Cedex 04, France.
Guillou
Vincent
V
François
Jean
J
Parrou
Jean-Luc
JL
D001692:Biological Transport; D003470:Culture Media; D015966:Gene Expression Regulation, Fungal; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D014198:Trehalase / Q000235:genetics / Q000378:metabolism; D014199:Trehalose / Q000378:metabolism*
D003470:Culture Media; D009004:Monosaccharide Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D027981:Symporters; C067575:maltose transport system, S cerevisiae; D014199:Trehalose; C097090:ATH1 protein, S cerevisiae; D014198:Trehalase