title: Probing Metabolite Space of Escherichia coli via Growth Medium Composition as Monitored by Two-Dimensional NMR Spectroscopy.
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.
authors.affiliation: Department of Chemistry, Sejong University, 209 Neungdong-Ro, Gwangjin-Gu, Seoul 143 - 747, Korea, (phone: +82-2-3408-3748; fax: +82-2-3408-4317). ykchae@sejong.ac.kr.
authors.forename: Young
authors.initials: Y
authors.lastname: Kee Chae
authors.affiliation: Department of Chemistry, Sejong University, 209 Neungdong-Ro, Gwangjin-Gu, Seoul 143 - 747, Korea, (phone: +82-2-3408-3748; fax: +82-2-3408-4317).
authors.forename: Seol
authors.initials: S
authors.lastname: Hyun Kim
authors.affiliation: Department of Chemistry, Sejong University, 209 Neungdong-Ro, Gwangjin-Gu, Seoul 143 - 747, Korea, (phone: +82-2-3408-3748; fax: +82-2-3408-4317).
authors.forename: Jin
authors.initials: J
authors.lastname: Seong Hyun
mesh_terms: D001622:Betaine / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D018698:Glutamic Acid / Q000032:analysis / Q000378:metabolism; D005978:Glutathione / Q000032:analysis / Q000378:metabolism; D005990:Glycerol / Q000032:analysis / Q000378:metabolism; D007930:Leucine / Q000032:analysis / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D011994:Recombinant Proteins; D018698:Glutamic Acid; D001622:Betaine; D005978:Glutathione; D007930:Leucine; D005990:Glycerol
keywords: Growth medium; Metabolite profiling; Multivariate analysis; NMR Spectroscopy