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Dynamic knockdown of E. coli central metabolism for redirecting fluxes of primary metabolites

Author(s)
Brockman, Irene Marie; Prather, Kristala L. Jones
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Abstract
Control of native enzyme levels is important when optimizing strains for overproduction of heterologous compounds. However, for many central metabolic enzymes, static knockdown results in poor growth and protein expression. We have developed a strategy for dynamically modulating the abundance of native enzymes within the host cell and applied this to a model system for myo-inositol production from glucose. This system relies on controlled degradation of a key glycolytic enzyme, phosphofructokinase-I (Pfk-I). Through tuning Pfk-I levels, we have been able to develop an Escherichia coli strain with a growth mode close to wild type and a production mode with an increased glucose-6-phosphate pool available for conversion into myo-inositol. The switch to production mode is trigged by inducer addition, allowing yield, titer, and productivity to be managed through induction time. By varying the time of Pfk-I degradation, we were able to achieve a two-fold improvement in yield and titers of myo-inositol.
Date issued
2014-12
URI
http://hdl.handle.net/1721.1/101771
Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Journal
Metabolic Engineering
Publisher
Elsevier
Citation
Brockman, Irene M., and Kristala L.J. Prather. “Dynamic Knockdown of E. Coli Central Metabolism for Redirecting Fluxes of Primary Metabolites.” Metabolic Engineering 28 (March 2015): 104–113.
Version: Author's final manuscript
ISSN
10967176
1096-7184

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