title: Comparative proteomics analysis of engineered Saccharomyces cerevisiae with enhanced biofuel precursor production.
abstract: 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.
authors.affiliation: School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Tang
authors.affiliation: School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Huixing
authors.initials: H
authors.lastname: Feng
authors.affiliation: School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Wei Ning
authors.initials: WN
authors.lastname: Chen
mesh_terms: D001275:ATP Citrate (pro-S)-Lyase / Q000378:metabolism; D056804:Biofuels; D002853:Chromatography, Liquid; D008401:Gas Chromatography-Mass Spectrometry; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D007521:Isocitrate Dehydrogenase / Q000235:genetics; D024101:Mitochondrial Proteins / Q000235:genetics; D040901:Proteomics / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D053719:Tandem Mass Spectrometry
chemical_list: D056804:Biofuels; D024101:Mitochondrial Proteins; D007521:Isocitrate Dehydrogenase; D001275:ATP Citrate (pro-S)-Lyase