Perez Marc, Luyten Kattie, Michel Remy, Riou Christine, Blondin Bruno
Equipe de Microbiologie, UMR Sciences Pour l'Oneologie, INRA-ENSAM-UMI, 2 place Viala, F-34060 Montpellier cedex 1, France.
FEMS Yeast Res. 2005 Feb;5(4-5):351-61. doi: 10.1016/j.femsyr.2004.09.005.
The transport of glucose and fructose into yeast cells is a critical step in the utilization of sugars during wine fermentation. Hexose uptake can be carried out by various Hxt carriers, each possessing distinct regulatory and transport-kinetic properties capable of influencing yeast fermentation capacity. We investigated the expression pattern of the hexose transporters Hxt1 to 7 at the promoter and protein levels in Saccharomyces cerevisiae during wine fermentation. The Hxt1p carrier was expressed only at the beginning of fermentation, and had no role during stationary phase. The Hxt3p carrier was the only one to be expressed throughout fermentation, displaying maximal expression at growth arrest and slowly decreasing in abundance over the course of the stationary phase. The high-affinity carriers Hxt6p and Hxt7p displayed similar expression profiles, with expression induced at entry into stationary phase and persisting throughout the phase. The expression of these two carriers occurred despite the presence of high amounts of hexoses, and the proteins were stably expressed when the cells were starved for nitrogen. The Hxt2p transporter was only transiently expressed during lag phase, which suggests a role for the protein in growth initiation. Characterization of glucose transport kinetics indicated the presence of a shift in the low-affinity component that is consistent with a predominant expression of Hxt1p during growth phase and of Hxt3p during stationary phase. In addition, a high-affinity uptake component consistent with functional expression of Hxt6p/Hxt7p was identified during stationary phase.
葡萄糖和果糖向酵母细胞内的转运是葡萄酒发酵过程中糖类利用的关键步骤。己糖的摄取可通过多种Hxt载体进行,每种载体都具有独特的调节和转运动力学特性,能够影响酵母的发酵能力。我们研究了酿酒酵母在葡萄酒发酵过程中己糖转运蛋白Hxt1至7在启动子和蛋白质水平的表达模式。Hxt1p载体仅在发酵开始时表达,在稳定期无作用。Hxt3p载体是整个发酵过程中唯一表达的载体,在生长停滞时表达量最高,在稳定期内丰度缓慢下降。高亲和力载体Hxt6p和Hxt7p表现出相似的表达谱,在进入稳定期时诱导表达并在整个阶段持续存在。尽管存在大量己糖,这两种载体仍会表达,并且当细胞缺乏氮时蛋白质会稳定表达。Hxt2p转运蛋白仅在延迟期短暂表达,这表明该蛋白在生长起始中起作用。葡萄糖转运动力学特征表明低亲和力成分存在转变,这与生长阶段Hxt1p的主要表达以及稳定期Hxt3p的主要表达一致。此外,在稳定期鉴定出了与Hxt6p/Hxt7p功能表达一致的高亲和力摄取成分。