Metabolic engineering of Escherichia coli for the production of succinate from glycerol |
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| Authors: | Matthew D Blankschien James M Clomburg Ramon Gonzalez |
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| Institution: | 1. Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, P.O. Box 1892, Houston, TX 77005, USA;2. Department of Bioengineering, Rice University, Houston, TX, USA;1. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan;2. Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, Hsinchu 30010, Taiwan;1. Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of Chemical Technology, Beijing 100029, China;2. Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China;3. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States;1. Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095, USA;2. Biomedical Engineering IDP, University of California, Los Angeles, CA 90095, USA];1. Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, USA;2. Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan;3. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan;4. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan;5. Material and Chemical Research Laboratories, Industrial Technology Research Institute, Hsinchu, Taiwan;1. State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Jinan 250100, China;2. School of Life Science, Linyi University, Linyi 276000, China;3. School of Energy and Environment, City University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong SAR |
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| Abstract: | Glycerol has become an ideal feedstock for the microbial production of bio-based chemicals due to its abundance, low cost, and high degree of reduction. We have previously reported the pathways and mechanisms for the utilization of glycerol by Escherichia coli in minimal salts medium under microaerobic conditions. Here we capitalize on such results to engineer E. coli for the production of value-added succinate from glycerol. Through metabolic engineering of E. coli metabolism, succinate production was greatly elevated by (1) blocking pathways for the synthesis of competing by-products lactate, ethanol, and acetate and (2) expressing Lactococcus lactis pyruvate carboxylase to drive the generation of succinate from the pyruvate node (as opposed to that of phosphoenolpyruvate). As such, these metabolic engineering strategies coupled cell growth to succinate production because the synthesis of succinate remained as the primary route of NAD+ regeneration. This feature enabled the operation of the succinate pathway in the absence of selective pressure (e.g. antibiotics). Our biocatalysts demonstrated a maximum specific productivity of ～400 mg succinate/gcell/h and a yield of 0.69 g succinate/g glycerol, on par with the use of glucose as a feedstock. |
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