The recombinant expression of proteins has been the method of choice to meet the demands from proteomics and structural genomics studies. Despite its successful production of many heterologous proteins, Escherichia coli failed to produce many other proteins in their native forms. This may be related to the fact that the stresses resulting from the overproduction interfere with cellular processes. To better understand the physiological change during the overproduction phase, we profiled the metabolites along the time course of the recombinant protein expression. We identified 32 metabolites collected from different time points in the protein production phase. The stress induced by protein production can be characterized by (A) the increased usage of aspartic acid, choline, glycerol, and N-acetyllysine; and (B) the accumulation of adenosine, alanine, oxidized glutathione, glycine, N-acetylputrescine, and uracil. We envision that this work can be used to create a strategy for the production of usable proteins in large quantities.

中文翻译：

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通过二维核磁共振波谱追踪重组蛋白表达过程中大肠杆菌的代谢足迹。


蛋白质的重组表达已成为满足蛋白质组学和结构基因组学研究需求的首选方法。尽管大肠杆菌成功产生了许多异源蛋白质，但它未能产生许多其他天然形式的蛋白质。这可能与过度生产产生的压力干扰细胞过程有关。为了更好地了解过量生产阶段的生理变化，我们分析了重组蛋白表达过程中的代谢物。我们鉴定了从蛋白质生产阶段的不同时间点收集的 32 种代谢物。蛋白质生产引起的应激可通过以下特征来表征：(A) 天冬氨酸、胆碱、甘油和 N-乙酰赖氨酸的使用量增加； (B) 腺苷、丙氨酸、氧化型谷胱甘肽、甘氨酸、N-乙酰腐胺和尿嘧啶的积累。我们设想这项工作可用于制定大量生产可用蛋白质的策略。