Abstract：
Reconstruction and optimization of biosynthetic pathways can help to overproduce target chemicals in microbial cell factories based on genetic engineering. However, the perturbation of biosynthetic pathways on cellular metabolism is not well investigated and profiling the engineered microbes remains challenging. The rapid development of omics tools has the potential to characterize the engineered microbial cell factory. Here, we performed label-free quantitative proteomic analysis and metabolomic analysis of engineered sabinene overproducing Saccharomyces cerevisiae strains. Combined metabolic analysis andproteomic analysis of targeted mevalonate (MVA) pathway showed that co-ordination of cytosolic and mitochondrial pathways had balanced metabolism, and genome integration of biosynthetic genes had higher sabinene production with less MVA enzymes. Furthermore, comparative proteomic analysis showed that compartmentalized mitochondria pathway had perturbation on central cellular metabolism. This study provided an omics analysis example for characterizing engineered cell factory, which can guide future regulation of the cellular metabolism and maintaining optimal protein expression levels for the synthesis of target products.
摘要：
生物合成途径的重建和优化有助于在基于基因工程的微生物细胞工厂中过量生产目标化学物质。然而,生物合成途径对细胞代谢的扰动没有得到很好的研究，对工程微生物的分析仍然具有挑战性。组学工具的快速发展具有表征工程化微生物细胞工厂的潜力。这里,我们进行了无标记的定量蛋白质组学分析和代谢组学分析工程sabinene过量生产酿酒酵母菌株.联合代谢分析和蛋白质组学分析靶向甲羟戊酸（MVA）途径显示，细胞溶质和线粒体途径的协调具有平衡的代谢，生物合成基因的基因组整合具有较高的sabinene产量，而MVA酶较少。此外，比较蛋白质组分析表明，线粒体通路对中枢细胞代谢有扰动。本研究为表征工程化细胞工厂提供了一个组学分析实例，这可以指导未来细胞代谢的调节和维持目标产物合成的最佳蛋白质表达水平。
