Tang Xiaoling, Feng Huixing, Zhang Jianhua, Chen Wei Ning
School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore, Singapore.
PLoS One. 2013 Dec 23;8(12):e84661. doi: 10.1371/journal.pone.0084661. eCollection 2013.
The yeast Saccharomyces cerevisiae was metabolically modified for enhanced biofuel precursor production by knocking out genes encoding mitochondrial isocitrate dehydrogenase and over-expression of a heterologous ATP-citrate lyase. A comparative iTRAQ-coupled 2D LC-MS/MS analysis was performed to obtain a global overview of ubiquitous protein expression changes in S. cerevisiae engineered strains. More than 300 proteins were identified. Among these proteins, 37 were found differentially expressed in engineered strains and they were classified into specific categories based on their enzyme functions. Most of the proteins involved in glycolytic and pyruvate branch-point pathways were found to be up-regulated and the proteins involved in respiration and glyoxylate pathway were however found to be down-regulated in engineered strains. Moreover, the metabolic modification of S. cerevisiae cells resulted in a number of up-regulated proteins involved in stress response and differentially expressed proteins involved in amino acid metabolism and protein biosynthesis pathways. These LC-MS/MS based proteomics analysis results not only offered extensive information in identifying potential protein-protein interactions, signal pathways and ubiquitous cellular changes elicited by the engineered pathways, but also provided a meaningful biological information platform serving further modification of yeast cells for enhanced biofuel production.
通过敲除编码线粒体异柠檬酸脱氢酶的基因并过表达异源ATP-柠檬酸裂解酶,对酿酒酵母进行代谢改造,以提高生物燃料前体的产量。进行了一项比较性的iTRAQ耦合二维液相色谱-串联质谱分析,以全面了解酿酒酵母工程菌株中普遍存在的蛋白质表达变化。鉴定出300多种蛋白质。在这些蛋白质中,有37种在工程菌株中差异表达,并根据其酶功能分为特定类别。发现参与糖酵解和丙酮酸分支点途径的大多数蛋白质上调,而参与呼吸和乙醛酸途径的蛋白质在工程菌株中下调。此外,酿酒酵母细胞的代谢改造导致许多参与应激反应的蛋白质上调,以及参与氨基酸代谢和蛋白质生物合成途径的差异表达蛋白质。这些基于液相色谱-串联质谱的蛋白质组学分析结果不仅在识别潜在的蛋白质-蛋白质相互作用、信号通路和工程途径引发的普遍细胞变化方面提供了广泛信息,还提供了一个有意义的生物学信息平台,用于进一步改造酵母细胞以提高生物燃料产量。