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Title: Inactivating FruR global regulator in plasmid-bearing Escherichia coli alters metabolic gene expression and improves growth rate
Authors: Ow, D.S.-W.
Lee, R.M.-Y.
Nissom, P.M.
Philp, R.
Oh, S.K.-W.
Yap, M.G.-S. 
Keywords: Catabolite repressor/activator
Fructose repressor
Global transcriptional regulator
Growth rate recovery
Metabolic engineering
Plasmid metabolic burden
Issue Date: 15-Sep-2007
Citation: Ow, D.S.-W., Lee, R.M.-Y., Nissom, P.M., Philp, R., Oh, S.K.-W., Yap, M.G.-S. (2007-09-15). Inactivating FruR global regulator in plasmid-bearing Escherichia coli alters metabolic gene expression and improves growth rate. Journal of Biotechnology 131 (3) : 261-269. ScholarBank@NUS Repository.
Abstract: The introduction of plasmids into Escherichia coli is known to impose a metabolic burden, which diminishes the growth rate. This effect could arise from perturbation of the central metabolic pathways, which supply precursors and energy for macromolecule synthesis. We knocked out a global regulator of central metabolism, FruR (also called Cra), to assess its phenotypic effect in E. coli carrying plasmids. During bioreactor runs, a higher specific growth rate of 0.91 h-1 was observed for the plasmid-bearing fruR knockout (P+ fruR) cells compared to its parental plasmid-bearing wildtype (P+ WT) cells (0.75 h-1), while both the plasmid-free cells displayed similar growth rates (1.0 h-1, respectively). To investigate gene expression changes possibly related to the growth rate recovery, quantitative reverse transcriptase PCR and 2DE proteomic studies were performed. In P+ fruR cells, expression of enzymes involved in sugar catabolism, glycolysis and transcription/translation processes were upregulated, while those related to gluconeogenesis, tricarboxylic acid cycle and stress response were downregulated. Our findings demonstrate that the inactivation of FruR global regulator in recombinant E. coli alters metabolic gene expression and significantly reduces growth retardation from the burden of maintaining a plasmid. This study represents the first attempt to explore the role of a global regulatory gene on plasmid metabolic burden. © 2007 Elsevier B.V. All rights reserved.
Source Title: Journal of Biotechnology
ISSN: 01681656
DOI: 10.1016/j.jbiotec.2007.07.508
Appears in Collections:Staff Publications

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