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http://dx.doi.org/10.5012/bkcs.2012.33.12.4041

Tracing Metabolite Footsteps of Escherichia coli Along the Time Course of Recombinant Protein Expression by Two-Dimensional NMR Spectroscopy  

Chae, Young Kee (Department of Chemistry and Institute for Chemical Biology, Sejong University)
Kim, Seol Hyun (Department of Chemistry and Institute for Chemical Biology, Sejong University)
Ellinger, James J. (Department of Biochemistry, University of Wisconsin-Madison)
Markley, John L. (Department of Biochemistry, University of Wisconsin-Madison)
Publication Information
Bulletin of the Korean Chemical Society / v.33, no.12, 2012 , pp. 4041-4046 More about this Journal
Abstract
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.
Keywords
Metabolite profiling; Recombinant protein; Overexpression; NMR;
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