Probing Metabolite Space of Escherichia coli via Growth Medium Composition as Monitored by Two-Dimensional NMR Spectroscopy.
Chem Biodivers. 2015 Jun;12(6):925-36
Authors: Kee Chae Y, Hyun Kim S, Seong Hyun J
Abstract
As for recombinant protein production, Escherichia coli is one of the most frequently employed hosts because it offers a simple and inexpensive, but rapid and high-yield system in addition to the vast information on its molecular genetics and biology. However, due to its prokaryotic nature, it often fails to produce eukaryotic proteins in a desired form. To devise a systematic way leading to a condition that produces a large amount of usable proteins, we attempted to monitor intracellular metabolites under various conditions, and to link them to recombinant protein production. With such an intention, we identified 31 metabolites from cells grown in different media by using two-dimensional (2D) NMR spectroscopy. Our results revealed that 1) the level of betaine was low, while that of glutamic acid was high when grown in minimal media; 2) the level of glycerol was constantly high in all cases; 3) the level of oxidized glutathione was lower in Luria broth (LB); and 4) the level of leucine was low in minimal media. We hope this work might shed light onto how to improve production of the target proteins by metabolite profiling.
[NMR images] Probing composition, sequence, and conformation
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2/12/2013 9:37:38 AM GMT
PROBING COMPOSITION, SEQUENCE, AND CONFORMATION
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Chem Biodivers. 2013 Oct;10(10):1816-1827
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NMR Spectroscopy has been established as a major tool for identification and quantification of metabolites in a living system. Since the metabolomics era began, one-dimensional NMR spectroscopy has been intensively employed due to its...
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Bull Korean Chem Soc. 2012 Dec 20;33(12):4041-4046
Authors: Chae YK, Kim SH, Ellinger JJ, Markley JL
Abstract
The recombinant expression of proteins has been the method of choice to meet the demands from proteomics and structural genomics...
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Authors: Yamasaki K, Taniguchi Y, Takeda N, Nakano K, Yamasaki T, Kanaya S, Oobatake M
Pressure denaturation of Escherichia coli ribonuclease HI (RNase HI) was studied by Fourier transform infrared (FTIR) and two-dimensional NMR...
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Eur J Biochem. 1996 Oct 15;241(2):489-97
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Biochemistry. 1994 May 10;33(18):5502-9
Authors: Hoeltzli SD, Frieden C
Escherichia coli dihydrofolate reductase contains five tryptophan residues distributed throughout its structure. In order to examine the regions of the protein surrounding these tryptophan...
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