A dynamic metabolite valve for the control of central carbon metabolism |
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| Authors: | Kevin V. Solomon Tarielle M. Sanders Kristala L.J. Prather |
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| Affiliation: | 1. Department of Chemical Engineering, Synthetic Biology Engineering Research Center (SynBERC), Massachusetts Institute of Technology, Cambridge, MA 02139, USA;2. Amgen Scholars Program, Department of Chemistry, Norfolk State University, Norfolk, VA 23504, USA;1. Laboratory for Bioinformatics, Graduate School of Systems Lifesciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan;2. Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita 565-0871, Osaka, Japan;3. Research Center for Transomics Medicine, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan;1. Interdisciplinary Bioinnovation PhD Program, Tulane University, New Orleans, LA, USA;2. Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, LA, USA;1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China |
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| Abstract: | Successful redirection of endogenous resources into heterologous pathways is a central tenet in the creation of efficient microbial cell factories. This redirection, however, may come at a price of poor biomass accumulation, reduced cofactor regeneration and low recombinant enzyme expression. In this study, we propose a metabolite valve to mitigate these issues by dynamically tuning endogenous processes to balance the demands of cell health and pathway efficiency. A control node of glucose utilization, glucokinase (Glk), was exogenously manipulated through either engineered antisense RNA or an inverting gene circuit. Using these techniques, we were able to directly control glycolytic flux, reducing the specific growth rate of engineered Escherichia coli by up to 50% without altering final biomass accumulation. This modulation was accompanied by successful redirection of glucose into a model pathway leading to an increase in the pathway yield and reduced carbon waste to acetate. This work represents one of the first examples of the dynamic redirection of glucose away from central carbon metabolism and enables the creation of novel, efficient intracellular pathways with glucose used directly as a substrate. |
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