Ando Akira, Tanaka Fumiko, Murata Yoshinori, Takagi Hiroshi, Shima Jun
National Food Research Institute, Ibaraki, Japan.
FEMS Yeast Res. 2006 Mar;6(2):249-67. doi: 10.1111/j.1567-1364.2006.00035.x.
Yeasts used in bread making are exposed to high concentrations of sucrose during sweet dough fermentation. Despite its importance, tolerance to high-sucrose stress is poorly understood at the gene level. To clarify the genes required for tolerance to high-sucrose stress, genome-wide screening was undertaken using the complete deletion strain collection of diploid Saccharomyces cerevisiae. The screening identified 273 deletions that yielded high sucrose sensitivity, approximately 20 of which were previously uncharacterized. These 273 deleted genes were classified based on their cellular function and localization of their gene products. Cross-sensitivity of the high-sucrose-sensitive mutants to high concentrations of NaCl and sorbitol was studied. Among the 273 sucrose-sensitive deletion mutants, 269 showed cross-sensitivities to sorbitol or NaCl, and four (i.e. ade5,7, ade6, ade8, and pde2) were specifically sensitive to high sucrose. The general stress response pathways via high-osmolarity glycerol and stress response element pathways and the function of the invertase in the ade mutants were similar to those in the wild-type strain. In the presence of high-sucrose stress, intracellular contents of ATP in ade mutants were at least twofold lower than that of the wild-type cells, suggesting that depletion of ATP is a factor in sensitivity to high-sucrose stress. The genes identified in this study might be important for tolerance to high-sucrose stress, and therefore should be target genes in future research into molecular modification for breeding of yeast tolerant to high-sucrose stress.
用于面包制作的酵母在甜面团发酵过程中会暴露于高浓度蔗糖环境中。尽管其重要性,但在基因水平上对高蔗糖胁迫耐受性的了解却很少。为了阐明耐受高蔗糖胁迫所需的基因,利用二倍体酿酒酵母的完全缺失菌株库进行了全基因组筛选。该筛选鉴定出273个缺失导致高蔗糖敏感性,其中约20个此前未被表征。这些273个缺失基因根据其细胞功能和基因产物的定位进行了分类。研究了高蔗糖敏感突变体对高浓度氯化钠和山梨醇的交叉敏感性。在这273个蔗糖敏感缺失突变体中,269个对山梨醇或氯化钠表现出交叉敏感性,四个(即ade5,7、ade6、ade8和pde2)对高蔗糖具有特异性敏感性。通过高渗甘油和应激反应元件途径的一般应激反应途径以及ade突变体中转化酶的功能与野生型菌株相似。在高蔗糖胁迫存在下,ade突变体中ATP的细胞内含量比野生型细胞至少低两倍,这表明ATP的消耗是对高蔗糖胁迫敏感的一个因素。本研究中鉴定出的基因可能对耐受高蔗糖胁迫很重要,因此应成为未来酵母耐高蔗糖胁迫分子改良育种研究的靶基因。