Shi Jiahua, Feng Huixing, Lee Jaslyn, Ning Chen Wei
School of Chemical and Biomedical Engineering, College of Engineering, Nanyang Technological University, Singapore.
PLoS One. 2013 Dec 26;8(12):e85532. doi: 10.1371/journal.pone.0085532. eCollection 2013.
Accumulation of intracellular lipid in oleaginous yeast cells has been studied for providing an alternative supply for energy, biofuel. Numerous studies have been conducted on increasing lipid content in oleaginous yeasts. However, few explore the mechanism of the high lipid accumulation ability of oleaginous yeast strains at the proteomics level. In this study, a time-course comparative proteomics analysis was introduced to compare the non-oleaginous yeast Saccharomyces cerevisiae, with two oleaginous yeast strains, Cryptococcus albidus and Rhodosporidium toruloides at different lipid accumulation stages. Two dimensional LC-MS/MS approach has been applied for protein profiling together with isobaric tag for relative and absolute quantitation (iTRAQ) labelling method. 132 proteins were identified when three yeast strains were all at early lipid accumulation stage; 122 and 116 proteins were found respectively within cells of three strains collected at middle and late lipid accumulation stages. Significantly up-regulation or down-regulation of proteins were experienced among comparison. Essential proteins correlated to lipid synthesis and regulation were detected. Our approach provides valuable indication and better understanding for lipid accumulation mechanism from proteomics level and would further contribute to genetic engineering of oleaginous yeasts.
为了提供能源、生物燃料的替代供应,人们对产油酵母细胞内脂质的积累进行了研究。关于提高产油酵母中脂质含量,已经开展了大量研究。然而,在蛋白质组学水平上,很少有人探索产油酵母菌株脂质高积累能力的机制。在本研究中,引入了时间进程比较蛋白质组学分析,以比较非产油酵母酿酒酵母与两种产油酵母菌株——浅白隐球酵母和红冬孢酵母在不同脂质积累阶段的情况。二维液相色谱-串联质谱法已与用于相对和绝对定量的等压标签(iTRAQ)标记方法一起应用于蛋白质谱分析。当三种酵母菌株都处于脂质积累早期阶段时,鉴定出了132种蛋白质;在脂质积累中期和后期收集的三种菌株的细胞中,分别发现了122种和116种蛋白质。在比较过程中,蛋白质出现了显著的上调或下调。检测到了与脂质合成和调控相关的关键蛋白质。我们的方法从蛋白质组学水平为脂质积累机制提供了有价值的线索和更好的理解,并将进一步有助于产油酵母的基因工程。