Cheng Jing-Sheng, Zhou Xiao, Ding Ming-Zhu, Yuan Ying-Jin
Tianjin University, People's Republic of China.
Appl Microbiol Biotechnol. 2009 Jul;83(5):909-23. doi: 10.1007/s00253-009-2037-1. Epub 2009 Jun 2.
The responses and adaptation mechanisms of the industrial Saccharomyces cerevisiae to vacuum fermentation were explored using proteomic approach. After qualitative and quantitative analyses, a total of 106 spots corresponding to 68 different proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The differentially expressed proteins were involved in amino acid and carbohydrate metabolisms, various signal pathways (Ras/MAPK, Ras-cyclic adenosine monophosphate, and HOG pathway), and heat shock and oxidative responses. Among them, alternations in levels of 17 proteins associated with carbohydrate metabolisms, in particular, the upregulations of proteins involved in glycolysis, trehalose biosynthesis, and the pentose phosphate pathway, suggested vacuum-induced redistribution of the metabolic fluxes. The upregulation of 17 heat stress and oxidative response proteins indicated that multifactors contributed to oxidative stresses by affecting cell redox homeostasis. Taken together with upregulation in 14-3-3 proteins levels, 22 proteins were detected in multispots, respectively, indicating that vacuum might have promoted posttranslational modifications of some proteins in S. cerevisiae. Further investigation revealed that the elevations of the differentially expressed proteins were mainly derived from vacuum stress rather than the absence of oxygen. These findings provide new molecular mechanisms for understanding of adaptation and tolerance of yeast to vacuum fermentation.
采用蛋白质组学方法探究了工业酿酒酵母对真空发酵的响应及适应机制。经过定性和定量分析,通过基质辅助激光解吸/电离飞行时间质谱法共鉴定出对应68种不同蛋白质的106个斑点。差异表达的蛋白质涉及氨基酸和碳水化合物代谢、各种信号通路(Ras/丝裂原活化蛋白激酶、Ras-环磷酸腺苷和高渗甘油途径)以及热休克和氧化应激反应。其中,17种与碳水化合物代谢相关的蛋白质水平发生变化,特别是参与糖酵解、海藻糖生物合成和磷酸戊糖途径的蛋白质上调,表明真空诱导了代谢通量的重新分配。17种热应激和氧化应激反应蛋白的上调表明,多种因素通过影响细胞氧化还原稳态导致氧化应激。结合14-3-3蛋白水平的上调,分别在多个斑点中检测到22种蛋白质,表明真空可能促进了酿酒酵母中某些蛋白质的翻译后修饰。进一步研究表明,差异表达蛋白的升高主要源于真空应激而非缺氧。这些发现为理解酵母对真空发酵的适应和耐受性提供了新的分子机制。