Construction and evolution of an Escherichia coli strain relying on nonoxidative glycolysis for sugar catabolism

Journal Article · Mon Mar 19 00:00:00 EDT 2018 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1802191115· OSTI ID:1426851

Lin, Paul P. ^[1]; Jaeger, Alec J. ^[1]; Wu, Tung-Yun ^[1]; Xu, Sharon C. ^[1]; Lee, Abraxa S. ^[1]; Gao, Fanke ^[1]; Chen, Po-Wei ^[1]; Liao, James C. ^[2]

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1. Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095,
2. Institute of Biological Chemistry, Academia Sinica, 115 Taipei, Taiwan

The Embden–Meyerhoff–Parnas (EMP) pathway, usually known as glycolysis, represents the fundamental biochemical infrastructure for sugar catabolism in almost all organisms, as it reveals key components for biosynthesis, energy metabolism, and global regulation. EMP-based metabolism synthesizes three-carbon (C3) metabolites before two-carbon (C2) metabolites and must emit one CO₂in the synthesis of the C2 building block, acetyl-CoA, a precursor for many industrially important products. Using rational design, genome editing, and evolution, here we replaced the native glycolytic pathways in Escherichia coli with the previously designed nonoxidative glycolysis (NOG), which bypasses initial C3 formation and directly generates stoichiometric amounts of C2 metabolites. The resulting strain, which contains 11 gene overexpressions, 10 gene deletions by design, and more than 50 genomic mutations (including 3 global regulators) through evolution, grows aerobically in glucose minimal medium but can ferment anaerobically to products with nearly complete carbon conservation. We confirmed that the strain metabolizes glucose through NOG by¹³C tracer experiments. This redesigned E. coli strain represents a different approach for carbon catabolism and may serve as a useful platform for bioproduction.

• Research Organization: Univ. of California, Los Angeles, CA (United States)
• Sponsoring Organization: USDOE Office of Science (SC), Biological and Environmental Research (BER)
• Grant/Contract Number: SC0012384; SC0006698
• OSTI ID: 1426851
• Alternate ID(s): OSTI ID: 1547357
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Vol. 115 Journal Issue: 14; ISSN 0027-8424
• Publisher: Proceedings of the National Academy of Sciences
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Escherichia coli
evolution
glycolysis
metabolic engineering
synthetic biology